RNA干扰技术抑制HGF表达对人结肠癌细胞SW620的增殖和移行行为的影响
摘要
结直肠癌是最常见的恶性肿瘤之一,占消化道恶性肿瘤的第二位,严重威胁到人类的健康。目前,临床上常用的结直肠癌的治疗方法如手术、放疗和化疗等虽然具有一定的疗效,但是创伤大,副反应重,预后差。因此,寻求一种新的、副反应小、治疗效果更好的方法势在必行。现已证实,结直肠癌的发展是一个多步骤、多阶段及多因素参与的疾病,而癌基因的变异及其参与的信号转导与肿瘤发展的关系极为密切。在众多信号传导网络中,蛋白酪氨酸激酶受体通路是最重要的传导通路之一。这也正是肝细胞生长因子(Hepatocyte Grouth Factor,HGF),及其受体c-Met的信号转导通路的首要环节。
肝细胞生长因子HGF又称分散因子( scatter factor , SF)是一种多肽生长因子,具有强促有丝分裂、组织器官成形、诱导上皮细胞迁移、侵袭以及诱导血管生成等作用。近年研究表明,在某些病理过程中,特别是在一些恶性肿瘤组织中包括结直肠癌在内,HGF/c- Met存在突变、过表达,而且与肿瘤发生、发展、转移及患者预后有密切关系。有研究表明,结直肠癌患者血清中HGF含量显著升高,其中合并肝转移的结直肠癌患者的血清中HGF含量较无肝转移的患者更高,推测HGF对结直肠癌的诊断、分期和预后的判断可能具有一定的价值,特别是对于结直肠癌患者有无肝转移将是一个很有价值的判断指标。因此,HGF极有可能成为新的结直肠癌治疗靶点。
RNA干扰(RNA interference, RNAi)是将与靶基因的mRNA同源互补的双链RNA(dsRNA )导入细胞,能特异性地降解该mRNA ,从而产生相应的功能表型缺失,属于转录后水平的基因沉默(post - transcriptional gene silence , PTGS)。目前,RNAi技术已经迅速而广泛的应用到基因功能鉴定、基因表达、转录后调控等热门研究领域,并为多种疾病的基因治疗提供了新的思路,尤其在肿瘤的基因治疗方面显示出诱人的前景。其中小分子干扰双链RNA(small interfering RNA , siRNA)是RNAi途径中的中间产物,它启动了细胞内RNAi的反应。有效的siRNA,能产生类似基因敲除的效果,为肿瘤的基因治疗开辟了新的途径。
我们前期实验已证明HGF可以促进人结肠癌细胞SW620的增殖和移行,并呈质量浓度依赖性。本实验采用RNAi方法抑制HGF基因表达,进一步探讨HGF对人结肠癌细胞株SW620的增殖和移行的影响,以期寻找治疗结直肠癌的新方法。
目的:探讨采用RNA干扰方法抑制HGF基因表达对结肠癌细胞SW620增殖、侵袭的影响。
方法:
1本实验用脂质体LIPOFECTAMINETM 2000介导,将Santa公司设计并合成的特异性的HGFα/βsiRNA转染入人结肠癌细胞株SW620,同时设立阴性对照和空白对照。在倒置荧光显微镜下测定细胞的瞬时转染效率。
2用Western Blot和RT-PCR方法检测细胞中HGF的mRNA和蛋白表达改变情况。
3倒置显微镜下观察各组细胞的形态学变化。
4 MTT检测细胞增殖活性的变化情况,并绘出各组细胞的生长曲线。
5流式细胞仪检测各组细胞的细胞周期变化。
6基底膜重建实验比较各组细胞的移行能力。
7扫描及透射电镜检测各组细胞的超微结构变化。
结果:
1倒置荧光显微镜下可观察到细胞瞬时转染效率约为70-80%。
2 Western Blot和RT-PCR方法检测转染HGFα/βsiRNA后SW620细胞中HGF的mRNA和蛋白表达均较对照组显著降低(p<0.05)。
3倒置显微镜下观察发现,在转染后48小时,转染HGFα/βsiRNA组,细胞增殖明显减少,细胞数量减少,细胞聚集情况明显抑制。
4 MTT结果显示,24、48、72和96小时四个时间段细胞的OD值,实验组与阴性对照组和空白对照组比较均有差异( p<0.05),实验组细胞增殖能力明显减弱,且效果在48小时最显著,此时其增殖能力抑制率>50%。
5流式细胞术结果显示,与阴性对照组和空白对照组相比,实验组中的G1期细胞明显增多,S期细胞减少。
6基底膜重建实验显示,实验组移行能力比阴性对照组降低58.2%,比空白对照组降低60.3%。
7扫描电子显微镜下观察到:实验组SW620细胞表面伪足成细丝状,突起数量减少。阴性对照组和正常对照组细胞可见粗壮的伪足,数目较多,有较明显的突起,且分布密集。
8透射电子显微镜观察SW620细胞的超微结构发现:RNA干扰后细胞内未见微管,只见少量微丝呈散在分布。
阴性对照组和正常对照组在核旁的胞质里可见较长较多的微管平行排列,微丝成束。
结论:应用RNA干扰技术抑制HGF的表达后明显抑制人结肠癌细胞株SW620的增殖和移行。
Colorectal carcinoma is the second common malignant tumor in digestive tract. Nowadays, the clinicle treatments such as operation, radiotherapy and therapy have some success on colorectal carcinoma. But, it has the big wound、bad adverse reaction and worse prognosis. Because of the younger of the colorectal carcinoma patients and the patient’s demand of high quality of life in post-treatment ,a new mothed which has the micro-wound、fewer adverse reaction even the radical cure of tumer is needed. It has been proved that the development of colorectal carcinoma included lots of steps, stages and facrors. Especially, the variation and the signal transduction of oncogene are closely associated with tumor progression and metastasis.One of the most important signal transductions is protein tyrosine kinase(PTK) ,which is the first link of the signaling of hepatocyte growth factor (HGF) and its acceptor c-Met .
HGF, also known as scatter factor (SF), is one of polypeptide growth factor (PGF), with pleiotropic effects clinicle treatments such as operation、radiotherapy and chemo have some success on colorectal carcinoma.But,it has the big wound、bad adverse reaction and worse prognosis.Because of the including mitogen, organogenesis, invasive growth and migration of endothelial cells, angiogenesis and so on[1].Recently, it has been shown that gene mutation and over-expression of HGF/c- Met are occurred in some procedure of pathology especially in some malignancies. Moreover, the mutation and over-expression of HGF/c- Met have a close relationship with tumor genesis, progression, metastasis and
prognosis. Udai found that the over-expression of HGF and c-Met was closely associated with tumer metastatic phenotype, metastases and survival. HGF will be worthy of diagnosing and deciding the stage and prognosis about colorectal carcinoma as a sign of serology. HGF was in the hope of finding a new therapeutic target to treat colorectal carcinoma.
RNA interference (RNAi) is a highly evolutionally conserved process of post-transcriptional gene silencing (PTGS) by which double stranded RNA (dsRNA), when introduced into a cell, causes sequence-specific degradation of homogolous mRNA sequences . Nowadays, RNAi causes specific gene silencing to take the place of traditional antisense nucleotide which is used in a hot wide variety of fields, including identification of gene function、regulation of post-transcriptional gene expression and so on. Additionally, RNAi provides the new way on gene therapy in kinds of disease especially makes the hot spot of gene therapy in tumor. As the intermediate product of RNAi, the small interfering RNA (siRNA) launched the RNAi in the cells and the result was known as knockout.
We had proved that HGF can promote the proliferation and migration of SW620 in a concentration-dependent manner previously . In this experiment, we would make down the gene expression of HGF by RNAi to investigate the effect of HGF on the proliferation and migration of SW620 in order to find a new therapeutic target of colorectal cancer.
Objective:To investigate the effects of the inhibition of HGF expression by RNAi on the proliferation and migration of SW620 .
Methods:
1 In this study, the specific HGFα/βsiRNA (h) designed and synthetized by Santa was transfected by lipidosome LIPOFECTAMINETM 2000 into SW620 . We also made negative control and blank group .The efficiency of cell transfection was examined by the inverted fluorescence microscope.
2 The expressions of HGF were analyzed by Western Blot and reverse-transcription PCR .
3 The morphology of SW620 was observed under the inverted microscope.
4 MTT was used to analyse the effect on the proliferation after RNAi and to draw the growth curve of each group.
5 The effect on the cell cycle after RNAi was determined by FCM.
6 The effect on the migration of cells after RNAi was studied by restoration of basal membrane.
7 The ultrastructure of SW620 after RNAi was observed under SEM and TEM.
Results:
1 The efficiency of cell transfection examined by the inverted fluorescence microscope was about 70-80%.
2 The expression of HGF in SW620 after the transfection of specific HGFα/βsiRNA was down- regulated significantly.
3 It can been seen under the inverted microscope that the proliferation and the quantity of cells were decreased apparently after 48h of the transfection of HGFα/βsiRNA.And cell aggregation was suppressed significantly.
4 The cell proliferation of HGFα/βsiRNA group was suppressed deeply as shown by MTT, Aditionaly, the most obvious effect happened at 48h.
5 The results of FCM showed that the cells of HGFα/βsiRNA group in G1 phase increased, but the cells in S phase decreased compared with the control groups.
6 As shown by restoration of basal membrane, the capability of migration of the cells of HGFα/βsiRNA group was degraded 58.2% than the negative control group and 60.3% than the untreated group.
7 The results of SEM showed that there were less parapodiums on the cell surface ,and the intensive and visible scabrosities can be seen after treated with HGFα/βsiRNA.However,the result of cells .in both control groups was converse.
8 As shown by TEM, the microtubules and microfilaments of SW620 decreased or disappeared emarkably after being treated with HGFα/βsiRNA.
Conclusion:The proliferation and migration of SW620 was suppressed obviously by RNAi of HGF .
肝细胞生长因子HGF又称分散因子( scatter factor , SF)是一种多肽生长因子,具有强促有丝分裂、组织器官成形、诱导上皮细胞迁移、侵袭以及诱导血管生成等作用。近年研究表明,在某些病理过程中,特别是在一些恶性肿瘤组织中包括结直肠癌在内,HGF/c- Met存在突变、过表达,而且与肿瘤发生、发展、转移及患者预后有密切关系。有研究表明,结直肠癌患者血清中HGF含量显著升高,其中合并肝转移的结直肠癌患者的血清中HGF含量较无肝转移的患者更高,推测HGF对结直肠癌的诊断、分期和预后的判断可能具有一定的价值,特别是对于结直肠癌患者有无肝转移将是一个很有价值的判断指标。因此,HGF极有可能成为新的结直肠癌治疗靶点。
RNA干扰(RNA interference, RNAi)是将与靶基因的mRNA同源互补的双链RNA(dsRNA )导入细胞,能特异性地降解该mRNA ,从而产生相应的功能表型缺失,属于转录后水平的基因沉默(post - transcriptional gene silence , PTGS)。目前,RNAi技术已经迅速而广泛的应用到基因功能鉴定、基因表达、转录后调控等热门研究领域,并为多种疾病的基因治疗提供了新的思路,尤其在肿瘤的基因治疗方面显示出诱人的前景。其中小分子干扰双链RNA(small interfering RNA , siRNA)是RNAi途径中的中间产物,它启动了细胞内RNAi的反应。有效的siRNA,能产生类似基因敲除的效果,为肿瘤的基因治疗开辟了新的途径。
我们前期实验已证明HGF可以促进人结肠癌细胞SW620的增殖和移行,并呈质量浓度依赖性。本实验采用RNAi方法抑制HGF基因表达,进一步探讨HGF对人结肠癌细胞株SW620的增殖和移行的影响,以期寻找治疗结直肠癌的新方法。
目的:探讨采用RNA干扰方法抑制HGF基因表达对结肠癌细胞SW620增殖、侵袭的影响。
方法:
1本实验用脂质体LIPOFECTAMINETM 2000介导,将Santa公司设计并合成的特异性的HGFα/βsiRNA转染入人结肠癌细胞株SW620,同时设立阴性对照和空白对照。在倒置荧光显微镜下测定细胞的瞬时转染效率。
2用Western Blot和RT-PCR方法检测细胞中HGF的mRNA和蛋白表达改变情况。
3倒置显微镜下观察各组细胞的形态学变化。
4 MTT检测细胞增殖活性的变化情况,并绘出各组细胞的生长曲线。
5流式细胞仪检测各组细胞的细胞周期变化。
6基底膜重建实验比较各组细胞的移行能力。
7扫描及透射电镜检测各组细胞的超微结构变化。
结果:
1倒置荧光显微镜下可观察到细胞瞬时转染效率约为70-80%。
2 Western Blot和RT-PCR方法检测转染HGFα/βsiRNA后SW620细胞中HGF的mRNA和蛋白表达均较对照组显著降低(p<0.05)。
3倒置显微镜下观察发现,在转染后48小时,转染HGFα/βsiRNA组,细胞增殖明显减少,细胞数量减少,细胞聚集情况明显抑制。
4 MTT结果显示,24、48、72和96小时四个时间段细胞的OD值,实验组与阴性对照组和空白对照组比较均有差异( p<0.05),实验组细胞增殖能力明显减弱,且效果在48小时最显著,此时其增殖能力抑制率>50%。
5流式细胞术结果显示,与阴性对照组和空白对照组相比,实验组中的G1期细胞明显增多,S期细胞减少。
6基底膜重建实验显示,实验组移行能力比阴性对照组降低58.2%,比空白对照组降低60.3%。
7扫描电子显微镜下观察到:实验组SW620细胞表面伪足成细丝状,突起数量减少。阴性对照组和正常对照组细胞可见粗壮的伪足,数目较多,有较明显的突起,且分布密集。
8透射电子显微镜观察SW620细胞的超微结构发现:RNA干扰后细胞内未见微管,只见少量微丝呈散在分布。
阴性对照组和正常对照组在核旁的胞质里可见较长较多的微管平行排列,微丝成束。
结论:应用RNA干扰技术抑制HGF的表达后明显抑制人结肠癌细胞株SW620的增殖和移行。
Colorectal carcinoma is the second common malignant tumor in digestive tract. Nowadays, the clinicle treatments such as operation, radiotherapy and therapy have some success on colorectal carcinoma. But, it has the big wound、bad adverse reaction and worse prognosis. Because of the younger of the colorectal carcinoma patients and the patient’s demand of high quality of life in post-treatment ,a new mothed which has the micro-wound、fewer adverse reaction even the radical cure of tumer is needed. It has been proved that the development of colorectal carcinoma included lots of steps, stages and facrors. Especially, the variation and the signal transduction of oncogene are closely associated with tumor progression and metastasis.One of the most important signal transductions is protein tyrosine kinase(PTK) ,which is the first link of the signaling of hepatocyte growth factor (HGF) and its acceptor c-Met .
HGF, also known as scatter factor (SF), is one of polypeptide growth factor (PGF), with pleiotropic effects clinicle treatments such as operation、radiotherapy and chemo have some success on colorectal carcinoma.But,it has the big wound、bad adverse reaction and worse prognosis.Because of the including mitogen, organogenesis, invasive growth and migration of endothelial cells, angiogenesis and so on[1].Recently, it has been shown that gene mutation and over-expression of HGF/c- Met are occurred in some procedure of pathology especially in some malignancies. Moreover, the mutation and over-expression of HGF/c- Met have a close relationship with tumor genesis, progression, metastasis and
prognosis. Udai found that the over-expression of HGF and c-Met was closely associated with tumer metastatic phenotype, metastases and survival. HGF will be worthy of diagnosing and deciding the stage and prognosis about colorectal carcinoma as a sign of serology. HGF was in the hope of finding a new therapeutic target to treat colorectal carcinoma.
RNA interference (RNAi) is a highly evolutionally conserved process of post-transcriptional gene silencing (PTGS) by which double stranded RNA (dsRNA), when introduced into a cell, causes sequence-specific degradation of homogolous mRNA sequences . Nowadays, RNAi causes specific gene silencing to take the place of traditional antisense nucleotide which is used in a hot wide variety of fields, including identification of gene function、regulation of post-transcriptional gene expression and so on. Additionally, RNAi provides the new way on gene therapy in kinds of disease especially makes the hot spot of gene therapy in tumor. As the intermediate product of RNAi, the small interfering RNA (siRNA) launched the RNAi in the cells and the result was known as knockout.
We had proved that HGF can promote the proliferation and migration of SW620 in a concentration-dependent manner previously . In this experiment, we would make down the gene expression of HGF by RNAi to investigate the effect of HGF on the proliferation and migration of SW620 in order to find a new therapeutic target of colorectal cancer.
Objective:To investigate the effects of the inhibition of HGF expression by RNAi on the proliferation and migration of SW620 .
Methods:
1 In this study, the specific HGFα/βsiRNA (h) designed and synthetized by Santa was transfected by lipidosome LIPOFECTAMINETM 2000 into SW620 . We also made negative control and blank group .The efficiency of cell transfection was examined by the inverted fluorescence microscope.
2 The expressions of HGF were analyzed by Western Blot and reverse-transcription PCR .
3 The morphology of SW620 was observed under the inverted microscope.
4 MTT was used to analyse the effect on the proliferation after RNAi and to draw the growth curve of each group.
5 The effect on the cell cycle after RNAi was determined by FCM.
6 The effect on the migration of cells after RNAi was studied by restoration of basal membrane.
7 The ultrastructure of SW620 after RNAi was observed under SEM and TEM.
Results:
1 The efficiency of cell transfection examined by the inverted fluorescence microscope was about 70-80%.
2 The expression of HGF in SW620 after the transfection of specific HGFα/βsiRNA was down- regulated significantly.
3 It can been seen under the inverted microscope that the proliferation and the quantity of cells were decreased apparently after 48h of the transfection of HGFα/βsiRNA.And cell aggregation was suppressed significantly.
4 The cell proliferation of HGFα/βsiRNA group was suppressed deeply as shown by MTT, Aditionaly, the most obvious effect happened at 48h.
5 The results of FCM showed that the cells of HGFα/βsiRNA group in G1 phase increased, but the cells in S phase decreased compared with the control groups.
6 As shown by restoration of basal membrane, the capability of migration of the cells of HGFα/βsiRNA group was degraded 58.2% than the negative control group and 60.3% than the untreated group.
7 The results of SEM showed that there were less parapodiums on the cell surface ,and the intensive and visible scabrosities can be seen after treated with HGFα/βsiRNA.However,the result of cells .in both control groups was converse.
8 As shown by TEM, the microtubules and microfilaments of SW620 decreased or disappeared emarkably after being treated with HGFα/βsiRNA.
Conclusion:The proliferation and migration of SW620 was suppressed obviously by RNAi of HGF .
引文
1 Hubbard SR , Mohammadi M, Schlessinger J .Autoregulatory mechanisms in protein-tyrosine kinases [J ] .J Biol Chem, 1998 ,273 (20) :11987-11990
2 Di Renzo MF, Olivero M, Giacomini A, et al. Overexpression and amplification of the met/HGF receptor geneduring the progression of colorectal cancer. Clin Cancer Res,1995, 1(2): 147-154
3李宏武,单吉贤.人大肠癌组织肝细胞生长因子及c-met的表达.世界华人消化杂志, 2004, 12(9): 2199-2201
4 Zeng Q , Chen S , You Z , et al . Hepatocyte growth factor inhibitsanoikis in head and neck squamous cell carcinoma cells by activation ofERKand Akt signaling independent of NF2κB[J] . J Biol Chem, 2002 ,277(28) : 25203-25208
5 Michalopoulos GK, DeFranees MC. Liver regeneration. Science, 1997, 276(5309): 60-66
6 Miyata Y, Ashida S, Nakamura T, et al. Over expression of hepatocyte growth factor receptor in renal carcinoma cells indirectly stimulates tumor growth in vivo. Biochemical and biophysical research communications, 2003, 302(4): 892-897
7 Udai S. Kammula, Eleanor J. Kuntz, Todd D. Francone, Zhaoshi Zeng, Molecular co-expression of the c-Met oncogene and hepatocyte growth factor in primary colon cancer predicts tumor stage and clinical outcome [J]Cancer Letters, 2007,248(2) : 219-228
8施为建,周巧云,蒋凤莲,鞠文东,王冬娥,陆小军血清肝细胞生长因子在大肠癌肝转移患者中的临床价值[J]临床肿瘤学杂志2007,12(11):822-824
9 Bottaro D P , Rubin J S , Faletto D L , et al. Identification of the hepatocyte growth factor receptor as the c2met proto2onco2 gene product [J]. Science , 1991 , 251(4995) : 802-804
10 Sawada K, Radjabi A R , Shinomiya N , et al. c2Met overex2 pression is a prognostic factor in ovarian cancer and an effec2 tive target for inhibition of peritonealdissemination and inva2 sion[J ]. Cancer Res , 2007 , 67 (4) : 1670-1679
11 Britta S, Kurt S. Differences in the migration capacity of primary human colon carcinoma cells (SW480) and their lymph node metastatic derivatives (SW620). Cancer Letters, 1998, 131(1):55-64
12 ElbashirSM,Harborth J ,Lendeckel W。etal .Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells, Nature, 2001, 411(6836):494-498
13 BrummelkampT R,Bernard R,Agami R .Stables uppressiono ftu morigenicity by virus-mediated RNA interference. Cancer Cell, 2002,2(3):243-247
14 Bernstein E,Caudy AA Hammond SM,etal. Role for a bidentate ribonuclease in the initiation step of RNA interferenc [J] .Nature,2001,409(6818):,363-366
15 Paranjpe S , Bowen W C , Bell A W, et al. Cell cycle effects resulting from inhibition of hepatocyte growth factor and its receptor c-Met in regenerating rat livers by RNA interference [J ] . Hepatology , 2007 , 45 (6) : 1471-1477
16 Lairen N .Bell,Liying Cai,Brian H.Johnstone, et al.A central role for hepatocyte growth factor in adipose tissue angiogenesis[J].Am J Physiol Endocrinol Metab,2008,294:336-344
17 Saiki A, Watanabe F, Murano T ,et al. Hepatocyte growth factor secreted by cultured adipocytes promotes tube formation of vascular endothelial cells in vitro. Int J Obes(Lond). 2006 ,30(11):1676-84
18 Paumelle R,Tulasne D,Kherrouche Z,et a1.Hepatocyte growth factor / scatter factor activates the ETS1 transcription factor by a RAS-RAF-MEK-ERK signaling pathway [J]. Oncogene,2002,21(15):2309-23l9
19 Ueda K , 1wahashi M , Matsuura I ,eta1 . Adenoviral-mediated gene transduction of the hepatocyte growth factor(HGF) antagonist,NK4,suppresses peritoneal metastases of gastric cancer in nude mice[J].Eur JCancer,2004,40 (14):2135-2142
20 Bray D. Cell Movement. 2nd ed. NewYork: Garland Publishing, 2001: 47-79
21 Plastino J, Sykes C. The actin sling shot. Curr Opin Cell Biol, 2005, 17(1): 62-66
22 Maulik G, Kijima T , Ma PC , et al . Modulation of the c-Met/Hepatocyte growth factor pathway in small cell lung cancer [J] . Clin Cancer Res , 2002 , 8 (2) :620-627.
23 Nobes C. D, Hall A, et al.Rho, rac and cdc42 GTPases: regulators of actin structures, cell adhesion and motility. Biochem. Soc. Trans., 1995,23: 456-459
24 Nobes C. D, Hawkins P, Stephens L, et al.Activation of the small GTP-binding proteins rho and rac by growth factor receptors. J. Cell Sci., 1995,108: 225-233
25 Irigoyen JP, Besser D, Nagamine Y. Cytoskeleton reorganization induces the urokinase-type plasminogen activator gene via the Ras/extracellular signal-regulatedkinase (ERK) signaling pathway. J Biol Chem. 1997 ,272(3):1904-9
26 Okamura, H, and Resh, M. D. p80/85 Cortactin Associates with the Src SH2 Domain and Colocalizes with v-Src in Transformed Cells. J. Biol. Chem.,1995 ,3(270): 26613-26618
27 Besser, D, Urich, M, Sakaue, M, et a1. Urokinase-type plasminogen activator gene regulation by polyomavirus middle-T antigen. ,Oncogene,1995 ,11(11):2383-91
28 Pulverer, B. J, Kyriakis, J. M, Avruch, J, et a1. Phosphorylation of c-jun mediated by MAP kinases. Nature. 1991 ,353(6345):670-4
29 Hunter T, Karin M.Cell. The regulation of transcription by phosphorylation. 1992,70(3):375-87 Ben-Ze'ev, A. Animal cell shape changes and gene expression. Bioessays, 1991, 13(5): 207-212
30 Ben-Ze'ev, A. Animal cell shape changes and gene expression. Bioessays, 1991, 13(5): 207-212
1 Zeng Q , Chen S , You Z , et al . Hepatocyte growth factor inhibitsanoikis in head and neck squamous cell carcinoma cells by activation ofERKand Akt signaling independent of NF kappa B[J] . J Biol Chem, 2002 ,277(28) : 25203-25208
2 Michalopoulos GK, DeFranees MC. Liver regeneration. Science, 1997, 276(5309): 60-66
3 Nakamura T. Strueture and function of hepatocyte growth factor. Progress in growth factor research, 1991, 3(l): 67-85
4 Stamos J , Lazarus R A , Yao X, et al. Crystal structure ofthe HGF betachain in complex with the Sema domain of the Met receptor[J]. EMBO J , 2004 , 23(12) : 2325-2335
5 Kirchhofer D , Lipari M T, Santell L , et al. Utilizing the ac- tivation mechanism of serine proteases to engineer hepatocyte growth factor into a Met antagonist [J]. Proc Natl Acad Sci USA , 2007 , 104(13) : 5306-5311
6 Furge KA,Zhang YW,Vande Woude GF.Met receptor tyrosine kinaseenhanced signaling through adapter protein[J].Oncogene,2000,19: 5582-5589
7 Ding S,Merkulova-Rainon T,Han ZC,et a1.HGF receptor up-regulation contributes to the angiogenic phenotype of human endothelial celsand promotes angiogenesis in vitro[J].Blood,2003,101:4816-4822
8 Sengupta S,Gherardi E,Sellers LA,et a1.Hepatocyte growth factor/seatter factor can induce angiogenesis independently of vascular endothelial growth factor[J].Arterioscler Thromb Vasc Biol,2003,23:69-75
9 Tokunou M,Niki T,Eguehi K,et a1.c-MET expression in myofibro-blasts : Role in autocrine activation and prognostic significance in lungadenoeareinoma[J].Am J Pathol,2001,158(4):1451-1463
10 Kajiya K, Hirakawa S, Ma B, et al. growth factor promotes lymphatic vessel formation and function. EMBO J, 2005, 24: 2885-95
11 Cao R, Bjorndahl MA, Gallego MI, et al. Hepatocyte growth factor is a lymphangiogenic factor with an indirect mechanism of action. Blood, 2006, 107: 3531-6
12 Goldman J, Rutkowski JM, Shields JD, et al. Cooperative and redundant roles of VEGFR-2 and VEGFR-3 signaling in adult lymphangiogenesis. FASEB J, 2007, 21: 1-10
13 Bottaro D P , Rubin J S , Faletto D L , et al. Identification of the hepatocyte growth factor receptor as the c2metproto2onco2 gene product [J]. Science , 1991 , 251(4995) : 802-804
14 Birchmeier C , Birchmeier W, Gherardi E , et al. Met , metastasis , motility and more [J]. Nat Rev Mol Cell Biol ,2003 , 4(12) : 915-925
15 Hubbard SR , Mohammadi M , Schlessinger J . Autoregulatory mechanisms in protein-tyrosine kinases[J].J Biol Chem,1998,273(20):11987-11990
16 Bardelli A ,Longati P,W illiams TA, et a1.A peptide representing the carboxyl-terminal tail of the met receptor inhibits kinase activity and invasive growth[J].J Biol Chem,1999,274(41):29274
17 Rosario M, Birchmeier W,et al. How to make tubes : signaling by the Met receptor tyrosine kinase[J]. Trends Cell Biol , 2003 ,13 (6) : 328-335
18 PaumeHe R,Tulasne D,Kherrouche Z,et a1.Hepatocyte growth factor / scatter factor activates the ETS1 transcription factor by a RAS-RAF-MEK-ERK signaling pathway [J]. Oncogene,2002,21(15):2309-23l9
19 Sawada K, Radjabi A R , Shinomiya N , et al. c-Met overexpression is a prognostic factor in ovarian cancer and an effective target for inhibition of peritoneal dissemination and invasion[J ]. Cancer Res , 2007 , 67 (4) : 1670-1679
20 Kermorgant S, Aparicio T, DessirierV, et al. Hepatocyte growth factor induces colonic cancer cell invasiveness viaenhanced mo2 tility and p rotease overp roduction [J]. Evidence for PI3 kinase and PKC involvement. Carcinogenesis, 2001, 22 ( 7 ) : 1035-1042
21董彤,辛晓燕.肝细胞生长因子及其受体c-Met在卵巢癌细胞中的表达[J].肿瘤与临床,2006,8(18):541-543.
22 Chongmei Liu, Zhiming Liu, Minzheng Ying, et a1.The significance of proto-oncogene HGF/SF receptor c-Met mRNA expression in nasopharyngeal carcinoma[J]. Chinese-German Journal of Clinical Oncology,2007,6( 3), 278-281
23 Strohmeyer D,Strauss F,Rossing c,et a1.Expression of bFGF, VEGF and c-met and their correlation with microvesse[density and progression in prostate carcinoma[J].Anticancer Res.2004,24(3):1797- 1804
24 Paranjpe S , Bowen W C , Bell A W, et al. Cell cycle effects resulting from inhibition of hepatocyte growth factor and its receptor c-Met in regenerating rat livers by RNA interference [J] . Hepatology , 2007 , 45 (6) : 1471-1477
25 H. Borgiel-Marek, D. Kajdaniuk, I. Niedzielska, et al. Daily oscillations of HGF in oral squamous cell carcinoma [J] . Journal of Cranio-Maxillofacial Surgery ,2008,36(1):186
26 S.U. Kang, Y.R. Yoon, H.S. Hwang,et al. c-Met gene amplification is associated with advanced stage colorectal cancer and liver metastases.[J]European Journal of CancerSupplements, 2007, 5(4): 328
27施为建,周巧云,蒋凤莲,等。血清肝细胞生长因子在大肠癌肝转移患者中的临床价值[J]临床肿瘤学杂志2007,12(11):822-824
28张玉华,吴文溪,魏尉,等。肝细胞生长因子及受体促进人大肠癌细胞血管内皮生长因子表达[J]南京医科大学学报(自然科学版),2007,27(3):216-219
29 Nie B, Shen Z, Wen JB, et al. AAV-HGFK1 and Ad-p53 cocktail therapy prolongs survival of mice with colon cancer, Mol Cancer Ther. 2008;7(9):2855-65
30 Kataoka H,Hamasuna K,Itoh H, et al. activation of hepatocyte growth factor/scatter Factor in colorectal carcinoma.Cancer Res 2000;60:6148-6159
31 Ueda K , 1wahashi M , Matsuura I ,eta1 . Adenoviral-mediated gene transduction of the hepatocyte growth factor(HGF) antagonist , NK4 ,suppresses peritoneal metastases of gastric cancer in nude mice[J].Eur JCancer,2004,40 (14):2135-2142
32 Udai S. Kammula, Eleanor J. Kuntz, Todd D. Francone, eta1. Molecular co-expression of the c-Met oncogene and hepatocyte growth factor in primary colon cancer predicts tumor stage and clinical outcome [J]Cancer Letters, 2007,248(2) : 219-228
33李宏武,梁健,张趣,肝细胞生长因子和MAPK途径在大肠癌细胞体外侵袭中的作用[J]中国医科大学学报,2006,35(6):606-608
34 Michela Fassetta , Lorenza D'Alessandro , Nadia Coltella , eta1. Hepatocyte growth factor installs a survival platform for colorectal cancer cell invasive growth and overcomes p38 MAPK mediated apoptosis, [J] Cellular Signalling , 2006,18(4):1967-1976
35 Date K, Matsumoto K, Shimura H , et al . HGF/ NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor [J] .FEBS Lett , 1997 ,420 (1) :1-6
36 Gakuhei Son, Tadamichi Hirano, Ekihiro Seki, eta1.Blockage of HGF/c-Met system by gene therapy (adenovirus-mediated NK4 gene) suppresses hepatocellular carcinoma in mice , [J] Journal of Hepatology ,2006,45(5):688-695
37 J. Wen, K. Matsumoto, N. Taniura, eta1.Hepatic gene expression of NK4, an HGF-antagonist/angiogenesis inhibitor,suppresses liver metastasis and invasive growth of colon cancer in mice, Cancer Gene Ther. 11 (2004):419–430
38 Davies G,Watkins G,Mason MD,et a1.Targeting the HGF/SF receptor c-met using a hammerhead ribozyme tran sgene reduces in vitro invasion and migration in prostate cancer cells[J].Prostate,2004,60(4):3l7-324
39 Kentaro Ueda, Makoto Iwahashi, Ichiro Matsuura, eta1. Adenoviral-mediated Gene transduction of the hepatocyte growth factor (HGF) antagonist, NK4, suppresses peritoneal metastases of gastric cancer in nude mice. [J]European Journal of Cancer, 2004,40(14) : 2135-2142
40 McManusMT, Sharp PA. Gene silencing in mammals by small interferingRNAs[J]. Nature, 2002, 3 (10) : 737-747
41 Elbashir SM, LendeckelW, Tuschi T. RNA interferenceis mediated by 212and 222nucleotide RNAs [J]. GenesDev, 2001, 15: 188-200
42 Zamore P D, Tuschl T, Sharp P A, et al. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals [J]. Cell, 2000, 101:25 - 33
43 Hutvagner G, Zamore P D. RNAi: nature abhors a double-strand[J]. CurrOp in GenetDev, 2002, 12: 225– 232
44 Sijen T, Fleenor J , Simmer F, et al. On the role of RNA amp lification in dsRNA2triggered gene silencing[J]. Cell,2001, 107: 465 - 476
45 Yu JY,DeRuiter SL, Turner DL, et al. RNA interference by exp ression of short2interfering RNAs and hairp in RNAs in mammalian cells [J].Proc Natl Acad Sci USA, 2002, 99 (9) : 6047-6052
46 Svoboda P, Stein P, Schultz RM, et al. RNAi in mouse oocytes and p reimp lantation embryos: effectiveness of hairp in dsRNA [J]. Biochem Biophys Res Commun, 2001, 287 (5) : 1099-1104
47 Ray KM, PaulA. RNA interference: from gene silencing to gene specific therapeutics[J]. Pharmacol Ther, 2005,107 (2) : 222-239
48 Wannenes F, Ciafre SA, Niola F, et al. Vector-based RNA interference against vascular endothelial growth factor-a significantly limits vascularization and growth of prostate cancer in vivo[J]. Cancer Gene Ther,2005, 12 (12) : 926-934
49 Yohsuke Matsumoto, Takahiro Motoki, Satoshi Kubota, et al. Inhibition of tumor– stromal interaction through HGF/Met signaling by valproic acid[J].Biochemical and Biophysical Research Communications, 2008,366(1):110-116
50 Wei-Jiunn Lee, Wen-Kang Chen, Chau-Jong Wang, et al. Apigenin inhibits HGF-promoted invasive growth and metastasis involvingblocking PI3K/Akt pathway andβ4 integrin function inMDA-MB-231 breast cancer cells[J]. Toxicology and Applied Pharmacology, 2008, 226(2) :178-191
51 M. Kong-Beltran, J. Stamos, D. Wickramasinghe, et al.The Sema domainof Met is necessary for receptor dimerization and activation . Cancer Cell,2004,6:75–84
52 Yuehai Shen, Qian Xie, Monica Norberg, et al.Geldanamycin derivative inhibition of HGF/SF-mediated Met tyrosine kinase receptor-dependent urokinase-plasminogen activation. Bioorganic & Medicinal Chemistry, 2005,13(16) : 4960-4971
53 Young Chang Lim, Hun Yi Park, Hye Sook Hwang, et al.(?)-Epigallocatechin-3-gallate (EGCG) inhibits HGF-induced invasion and metastasis in hypopharyngeal carcinoma cells [J ] Cancer Letters, 2008,271(1): 140-152
54 Wang SY,Chen B,Zhan YQ,el a1. SU5416 is a potent inhibitor of hepatocyte growth factor receptor (c-M et) and blocks HGF induced invasiveness of human HepG2 hepatomacells[J].J Hepatol,2004,41(2):267-273
55 M ichieli P , M azzone M , Basilico C . et a1. Targeting the tumor and its microenvironment by a dual—function decoy Met receptor[J].Cancer Cell,2004,6(1):6l-73
56 Sugano M, Iwasaki Y, Abe M, TNF-alpha employs a protein-tyrosine phosphatase to inhibit activation of hepatocyte growth factor receptor and hepatocyte growth factor-induced endothelial cell proliferation, Mol Cell Biochem, 2009 ,322(1-2):113-117
2 Di Renzo MF, Olivero M, Giacomini A, et al. Overexpression and amplification of the met/HGF receptor geneduring the progression of colorectal cancer. Clin Cancer Res,1995, 1(2): 147-154
3李宏武,单吉贤.人大肠癌组织肝细胞生长因子及c-met的表达.世界华人消化杂志, 2004, 12(9): 2199-2201
4 Zeng Q , Chen S , You Z , et al . Hepatocyte growth factor inhibitsanoikis in head and neck squamous cell carcinoma cells by activation ofERKand Akt signaling independent of NF2κB[J] . J Biol Chem, 2002 ,277(28) : 25203-25208
5 Michalopoulos GK, DeFranees MC. Liver regeneration. Science, 1997, 276(5309): 60-66
6 Miyata Y, Ashida S, Nakamura T, et al. Over expression of hepatocyte growth factor receptor in renal carcinoma cells indirectly stimulates tumor growth in vivo. Biochemical and biophysical research communications, 2003, 302(4): 892-897
7 Udai S. Kammula, Eleanor J. Kuntz, Todd D. Francone, Zhaoshi Zeng, Molecular co-expression of the c-Met oncogene and hepatocyte growth factor in primary colon cancer predicts tumor stage and clinical outcome [J]Cancer Letters, 2007,248(2) : 219-228
8施为建,周巧云,蒋凤莲,鞠文东,王冬娥,陆小军血清肝细胞生长因子在大肠癌肝转移患者中的临床价值[J]临床肿瘤学杂志2007,12(11):822-824
9 Bottaro D P , Rubin J S , Faletto D L , et al. Identification of the hepatocyte growth factor receptor as the c2met proto2onco2 gene product [J]. Science , 1991 , 251(4995) : 802-804
10 Sawada K, Radjabi A R , Shinomiya N , et al. c2Met overex2 pression is a prognostic factor in ovarian cancer and an effec2 tive target for inhibition of peritonealdissemination and inva2 sion[J ]. Cancer Res , 2007 , 67 (4) : 1670-1679
11 Britta S, Kurt S. Differences in the migration capacity of primary human colon carcinoma cells (SW480) and their lymph node metastatic derivatives (SW620). Cancer Letters, 1998, 131(1):55-64
12 ElbashirSM,Harborth J ,Lendeckel W。etal .Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells, Nature, 2001, 411(6836):494-498
13 BrummelkampT R,Bernard R,Agami R .Stables uppressiono ftu morigenicity by virus-mediated RNA interference. Cancer Cell, 2002,2(3):243-247
14 Bernstein E,Caudy AA Hammond SM,etal. Role for a bidentate ribonuclease in the initiation step of RNA interferenc [J] .Nature,2001,409(6818):,363-366
15 Paranjpe S , Bowen W C , Bell A W, et al. Cell cycle effects resulting from inhibition of hepatocyte growth factor and its receptor c-Met in regenerating rat livers by RNA interference [J ] . Hepatology , 2007 , 45 (6) : 1471-1477
16 Lairen N .Bell,Liying Cai,Brian H.Johnstone, et al.A central role for hepatocyte growth factor in adipose tissue angiogenesis[J].Am J Physiol Endocrinol Metab,2008,294:336-344
17 Saiki A, Watanabe F, Murano T ,et al. Hepatocyte growth factor secreted by cultured adipocytes promotes tube formation of vascular endothelial cells in vitro. Int J Obes(Lond). 2006 ,30(11):1676-84
18 Paumelle R,Tulasne D,Kherrouche Z,et a1.Hepatocyte growth factor / scatter factor activates the ETS1 transcription factor by a RAS-RAF-MEK-ERK signaling pathway [J]. Oncogene,2002,21(15):2309-23l9
19 Ueda K , 1wahashi M , Matsuura I ,eta1 . Adenoviral-mediated gene transduction of the hepatocyte growth factor(HGF) antagonist,NK4,suppresses peritoneal metastases of gastric cancer in nude mice[J].Eur JCancer,2004,40 (14):2135-2142
20 Bray D. Cell Movement. 2nd ed. NewYork: Garland Publishing, 2001: 47-79
21 Plastino J, Sykes C. The actin sling shot. Curr Opin Cell Biol, 2005, 17(1): 62-66
22 Maulik G, Kijima T , Ma PC , et al . Modulation of the c-Met/Hepatocyte growth factor pathway in small cell lung cancer [J] . Clin Cancer Res , 2002 , 8 (2) :620-627.
23 Nobes C. D, Hall A, et al.Rho, rac and cdc42 GTPases: regulators of actin structures, cell adhesion and motility. Biochem. Soc. Trans., 1995,23: 456-459
24 Nobes C. D, Hawkins P, Stephens L, et al.Activation of the small GTP-binding proteins rho and rac by growth factor receptors. J. Cell Sci., 1995,108: 225-233
25 Irigoyen JP, Besser D, Nagamine Y. Cytoskeleton reorganization induces the urokinase-type plasminogen activator gene via the Ras/extracellular signal-regulatedkinase (ERK) signaling pathway. J Biol Chem. 1997 ,272(3):1904-9
26 Okamura, H, and Resh, M. D. p80/85 Cortactin Associates with the Src SH2 Domain and Colocalizes with v-Src in Transformed Cells. J. Biol. Chem.,1995 ,3(270): 26613-26618
27 Besser, D, Urich, M, Sakaue, M, et a1. Urokinase-type plasminogen activator gene regulation by polyomavirus middle-T antigen. ,Oncogene,1995 ,11(11):2383-91
28 Pulverer, B. J, Kyriakis, J. M, Avruch, J, et a1. Phosphorylation of c-jun mediated by MAP kinases. Nature. 1991 ,353(6345):670-4
29 Hunter T, Karin M.Cell. The regulation of transcription by phosphorylation. 1992,70(3):375-87 Ben-Ze'ev, A. Animal cell shape changes and gene expression. Bioessays, 1991, 13(5): 207-212
30 Ben-Ze'ev, A. Animal cell shape changes and gene expression. Bioessays, 1991, 13(5): 207-212
1 Zeng Q , Chen S , You Z , et al . Hepatocyte growth factor inhibitsanoikis in head and neck squamous cell carcinoma cells by activation ofERKand Akt signaling independent of NF kappa B[J] . J Biol Chem, 2002 ,277(28) : 25203-25208
2 Michalopoulos GK, DeFranees MC. Liver regeneration. Science, 1997, 276(5309): 60-66
3 Nakamura T. Strueture and function of hepatocyte growth factor. Progress in growth factor research, 1991, 3(l): 67-85
4 Stamos J , Lazarus R A , Yao X, et al. Crystal structure ofthe HGF betachain in complex with the Sema domain of the Met receptor[J]. EMBO J , 2004 , 23(12) : 2325-2335
5 Kirchhofer D , Lipari M T, Santell L , et al. Utilizing the ac- tivation mechanism of serine proteases to engineer hepatocyte growth factor into a Met antagonist [J]. Proc Natl Acad Sci USA , 2007 , 104(13) : 5306-5311
6 Furge KA,Zhang YW,Vande Woude GF.Met receptor tyrosine kinaseenhanced signaling through adapter protein[J].Oncogene,2000,19: 5582-5589
7 Ding S,Merkulova-Rainon T,Han ZC,et a1.HGF receptor up-regulation contributes to the angiogenic phenotype of human endothelial celsand promotes angiogenesis in vitro[J].Blood,2003,101:4816-4822
8 Sengupta S,Gherardi E,Sellers LA,et a1.Hepatocyte growth factor/seatter factor can induce angiogenesis independently of vascular endothelial growth factor[J].Arterioscler Thromb Vasc Biol,2003,23:69-75
9 Tokunou M,Niki T,Eguehi K,et a1.c-MET expression in myofibro-blasts : Role in autocrine activation and prognostic significance in lungadenoeareinoma[J].Am J Pathol,2001,158(4):1451-1463
10 Kajiya K, Hirakawa S, Ma B, et al. growth factor promotes lymphatic vessel formation and function. EMBO J, 2005, 24: 2885-95
11 Cao R, Bjorndahl MA, Gallego MI, et al. Hepatocyte growth factor is a lymphangiogenic factor with an indirect mechanism of action. Blood, 2006, 107: 3531-6
12 Goldman J, Rutkowski JM, Shields JD, et al. Cooperative and redundant roles of VEGFR-2 and VEGFR-3 signaling in adult lymphangiogenesis. FASEB J, 2007, 21: 1-10
13 Bottaro D P , Rubin J S , Faletto D L , et al. Identification of the hepatocyte growth factor receptor as the c2metproto2onco2 gene product [J]. Science , 1991 , 251(4995) : 802-804
14 Birchmeier C , Birchmeier W, Gherardi E , et al. Met , metastasis , motility and more [J]. Nat Rev Mol Cell Biol ,2003 , 4(12) : 915-925
15 Hubbard SR , Mohammadi M , Schlessinger J . Autoregulatory mechanisms in protein-tyrosine kinases[J].J Biol Chem,1998,273(20):11987-11990
16 Bardelli A ,Longati P,W illiams TA, et a1.A peptide representing the carboxyl-terminal tail of the met receptor inhibits kinase activity and invasive growth[J].J Biol Chem,1999,274(41):29274
17 Rosario M, Birchmeier W,et al. How to make tubes : signaling by the Met receptor tyrosine kinase[J]. Trends Cell Biol , 2003 ,13 (6) : 328-335
18 PaumeHe R,Tulasne D,Kherrouche Z,et a1.Hepatocyte growth factor / scatter factor activates the ETS1 transcription factor by a RAS-RAF-MEK-ERK signaling pathway [J]. Oncogene,2002,21(15):2309-23l9
19 Sawada K, Radjabi A R , Shinomiya N , et al. c-Met overexpression is a prognostic factor in ovarian cancer and an effective target for inhibition of peritoneal dissemination and invasion[J ]. Cancer Res , 2007 , 67 (4) : 1670-1679
20 Kermorgant S, Aparicio T, DessirierV, et al. Hepatocyte growth factor induces colonic cancer cell invasiveness viaenhanced mo2 tility and p rotease overp roduction [J]. Evidence for PI3 kinase and PKC involvement. Carcinogenesis, 2001, 22 ( 7 ) : 1035-1042
21董彤,辛晓燕.肝细胞生长因子及其受体c-Met在卵巢癌细胞中的表达[J].肿瘤与临床,2006,8(18):541-543.
22 Chongmei Liu, Zhiming Liu, Minzheng Ying, et a1.The significance of proto-oncogene HGF/SF receptor c-Met mRNA expression in nasopharyngeal carcinoma[J]. Chinese-German Journal of Clinical Oncology,2007,6( 3), 278-281
23 Strohmeyer D,Strauss F,Rossing c,et a1.Expression of bFGF, VEGF and c-met and their correlation with microvesse[density and progression in prostate carcinoma[J].Anticancer Res.2004,24(3):1797- 1804
24 Paranjpe S , Bowen W C , Bell A W, et al. Cell cycle effects resulting from inhibition of hepatocyte growth factor and its receptor c-Met in regenerating rat livers by RNA interference [J] . Hepatology , 2007 , 45 (6) : 1471-1477
25 H. Borgiel-Marek, D. Kajdaniuk, I. Niedzielska, et al. Daily oscillations of HGF in oral squamous cell carcinoma [J] . Journal of Cranio-Maxillofacial Surgery ,2008,36(1):186
26 S.U. Kang, Y.R. Yoon, H.S. Hwang,et al. c-Met gene amplification is associated with advanced stage colorectal cancer and liver metastases.[J]European Journal of CancerSupplements, 2007, 5(4): 328
27施为建,周巧云,蒋凤莲,等。血清肝细胞生长因子在大肠癌肝转移患者中的临床价值[J]临床肿瘤学杂志2007,12(11):822-824
28张玉华,吴文溪,魏尉,等。肝细胞生长因子及受体促进人大肠癌细胞血管内皮生长因子表达[J]南京医科大学学报(自然科学版),2007,27(3):216-219
29 Nie B, Shen Z, Wen JB, et al. AAV-HGFK1 and Ad-p53 cocktail therapy prolongs survival of mice with colon cancer, Mol Cancer Ther. 2008;7(9):2855-65
30 Kataoka H,Hamasuna K,Itoh H, et al. activation of hepatocyte growth factor/scatter Factor in colorectal carcinoma.Cancer Res 2000;60:6148-6159
31 Ueda K , 1wahashi M , Matsuura I ,eta1 . Adenoviral-mediated gene transduction of the hepatocyte growth factor(HGF) antagonist , NK4 ,suppresses peritoneal metastases of gastric cancer in nude mice[J].Eur JCancer,2004,40 (14):2135-2142
32 Udai S. Kammula, Eleanor J. Kuntz, Todd D. Francone, eta1. Molecular co-expression of the c-Met oncogene and hepatocyte growth factor in primary colon cancer predicts tumor stage and clinical outcome [J]Cancer Letters, 2007,248(2) : 219-228
33李宏武,梁健,张趣,肝细胞生长因子和MAPK途径在大肠癌细胞体外侵袭中的作用[J]中国医科大学学报,2006,35(6):606-608
34 Michela Fassetta , Lorenza D'Alessandro , Nadia Coltella , eta1. Hepatocyte growth factor installs a survival platform for colorectal cancer cell invasive growth and overcomes p38 MAPK mediated apoptosis, [J] Cellular Signalling , 2006,18(4):1967-1976
35 Date K, Matsumoto K, Shimura H , et al . HGF/ NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor [J] .FEBS Lett , 1997 ,420 (1) :1-6
36 Gakuhei Son, Tadamichi Hirano, Ekihiro Seki, eta1.Blockage of HGF/c-Met system by gene therapy (adenovirus-mediated NK4 gene) suppresses hepatocellular carcinoma in mice , [J] Journal of Hepatology ,2006,45(5):688-695
37 J. Wen, K. Matsumoto, N. Taniura, eta1.Hepatic gene expression of NK4, an HGF-antagonist/angiogenesis inhibitor,suppresses liver metastasis and invasive growth of colon cancer in mice, Cancer Gene Ther. 11 (2004):419–430
38 Davies G,Watkins G,Mason MD,et a1.Targeting the HGF/SF receptor c-met using a hammerhead ribozyme tran sgene reduces in vitro invasion and migration in prostate cancer cells[J].Prostate,2004,60(4):3l7-324
39 Kentaro Ueda, Makoto Iwahashi, Ichiro Matsuura, eta1. Adenoviral-mediated Gene transduction of the hepatocyte growth factor (HGF) antagonist, NK4, suppresses peritoneal metastases of gastric cancer in nude mice. [J]European Journal of Cancer, 2004,40(14) : 2135-2142
40 McManusMT, Sharp PA. Gene silencing in mammals by small interferingRNAs[J]. Nature, 2002, 3 (10) : 737-747
41 Elbashir SM, LendeckelW, Tuschi T. RNA interferenceis mediated by 212and 222nucleotide RNAs [J]. GenesDev, 2001, 15: 188-200
42 Zamore P D, Tuschl T, Sharp P A, et al. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals [J]. Cell, 2000, 101:25 - 33
43 Hutvagner G, Zamore P D. RNAi: nature abhors a double-strand[J]. CurrOp in GenetDev, 2002, 12: 225– 232
44 Sijen T, Fleenor J , Simmer F, et al. On the role of RNA amp lification in dsRNA2triggered gene silencing[J]. Cell,2001, 107: 465 - 476
45 Yu JY,DeRuiter SL, Turner DL, et al. RNA interference by exp ression of short2interfering RNAs and hairp in RNAs in mammalian cells [J].Proc Natl Acad Sci USA, 2002, 99 (9) : 6047-6052
46 Svoboda P, Stein P, Schultz RM, et al. RNAi in mouse oocytes and p reimp lantation embryos: effectiveness of hairp in dsRNA [J]. Biochem Biophys Res Commun, 2001, 287 (5) : 1099-1104
47 Ray KM, PaulA. RNA interference: from gene silencing to gene specific therapeutics[J]. Pharmacol Ther, 2005,107 (2) : 222-239
48 Wannenes F, Ciafre SA, Niola F, et al. Vector-based RNA interference against vascular endothelial growth factor-a significantly limits vascularization and growth of prostate cancer in vivo[J]. Cancer Gene Ther,2005, 12 (12) : 926-934
49 Yohsuke Matsumoto, Takahiro Motoki, Satoshi Kubota, et al. Inhibition of tumor– stromal interaction through HGF/Met signaling by valproic acid[J].Biochemical and Biophysical Research Communications, 2008,366(1):110-116
50 Wei-Jiunn Lee, Wen-Kang Chen, Chau-Jong Wang, et al. Apigenin inhibits HGF-promoted invasive growth and metastasis involvingblocking PI3K/Akt pathway andβ4 integrin function inMDA-MB-231 breast cancer cells[J]. Toxicology and Applied Pharmacology, 2008, 226(2) :178-191
51 M. Kong-Beltran, J. Stamos, D. Wickramasinghe, et al.The Sema domainof Met is necessary for receptor dimerization and activation . Cancer Cell,2004,6:75–84
52 Yuehai Shen, Qian Xie, Monica Norberg, et al.Geldanamycin derivative inhibition of HGF/SF-mediated Met tyrosine kinase receptor-dependent urokinase-plasminogen activation. Bioorganic & Medicinal Chemistry, 2005,13(16) : 4960-4971
53 Young Chang Lim, Hun Yi Park, Hye Sook Hwang, et al.(?)-Epigallocatechin-3-gallate (EGCG) inhibits HGF-induced invasion and metastasis in hypopharyngeal carcinoma cells [J ] Cancer Letters, 2008,271(1): 140-152
54 Wang SY,Chen B,Zhan YQ,el a1. SU5416 is a potent inhibitor of hepatocyte growth factor receptor (c-M et) and blocks HGF induced invasiveness of human HepG2 hepatomacells[J].J Hepatol,2004,41(2):267-273
55 M ichieli P , M azzone M , Basilico C . et a1. Targeting the tumor and its microenvironment by a dual—function decoy Met receptor[J].Cancer Cell,2004,6(1):6l-73
56 Sugano M, Iwasaki Y, Abe M, TNF-alpha employs a protein-tyrosine phosphatase to inhibit activation of hepatocyte growth factor receptor and hepatocyte growth factor-induced endothelial cell proliferation, Mol Cell Biochem, 2009 ,322(1-2):113-117