特殊富含AT序列结合蛋白1在胃癌组织中的表达及其对胃癌细胞增殖的影响
|
摘要
目的:研究特殊富含AT序列结合蛋白1(SATB1)在胃癌组织中的表达及其对胃癌细胞增殖的影响。方法:免疫组化法检测30例胃癌组织、淋巴结组织及癌旁组织中SATB1的表达; RT-PCR及Western blot检测胃癌细胞株MGC-803和正常胃黏膜永生化细胞GES-1中SATB1的mRNA和蛋白的含量;电转染技术将真核表达质粒pc DNA3. 1-His-SATB1(HisSATB1)、载体质粒pc DNA3. 1(His-N)、沉默质粒p PLK+Puro-SATB1 shRNA(sh-SATB1)、沉默载体质粒p PLK+Puro(sh-N)转染入MGC-803细胞,24 h后Western blot检测各组细胞中SATB1、AKT、p-AKT的表达; CCK8检测细胞增殖活性。结果:SATB1在胃癌组织、转移淋巴结组织及MGC-803细胞中的表达水平显著升高(P<0. 05)。sh-SATB1组MGC-803细胞中SATB1的蛋白表达显著降低,细胞增殖活性、AKT、p-AKT蛋白表达均显著降低(P<0. 05); His-SATB1组MGC-803细胞中SATB1蛋白表达水平显著升高,细胞增殖活性增强,AKT、p-AKT蛋白表达均显著升高(P<0. 05)。结论:SATB1在胃癌组织及细胞中高表达,SATB1可促进胃癌细胞的增殖,该效应可能是通过调控AKT合成及激活PI3K/AKT信号通路实现的。
Objective: To study the expression of special AT rich sequence binding protein 1( SATB1) in gastric cancer tissues and its effect on the proliferation of gastric cancer cells. Methods: The expression of SATB1 was detected by immunohistochemistry in30 cases of gastric cancer,lymph node and paracancerous tissues. The mRNA and protein expression of SATB1 were detected by RTPCR and Western blot in gastric cancer cell line MGC-803 and normal gastric mucosal immortalized cell GES-1. Eukaryotic expression plasmid pc DNA3. 1-His-SATB1( His-SATB1),vector plasmid pc DNA3. 1( His-N),silencing plasmid p PLK+Puro-SATB1 shRNA( shSATB1),silencing vector plasmid p PLK+Puro( sh-N) were transfected into MGC-803 cells using electroporation technology,and after24 h Western blot was used to determine the expression of SATB1,AKT and p-AKT in each cell group. Cell proliferation was detected by CCK8. Results: The expression of SATB1 in gastric cancer tissues,metastatic lymph node tissues and MGC-803 cells significantly increased( P< 0. 05). The protein expression of SATB1 remarkably decreased in sh-SATB1 group,with decreased cell proliferation activity,AKT and p-AKT protein expression( P<0. 05). The protein expression of SATB1 in His-SATB1 group significantly increased,with enhanced cell proliferation,increased AKT and p-AKT protein expression( P<0. 05). Conclusion: SATB1 is highly expressed in gastric cancer tissues and cells. SATB1 promotes the proliferation of gastric cancer cells,which could be achieved by regulating AKT synthesis and activating PI3 K/AKT signaling pathway.
Objective: To study the expression of special AT rich sequence binding protein 1( SATB1) in gastric cancer tissues and its effect on the proliferation of gastric cancer cells. Methods: The expression of SATB1 was detected by immunohistochemistry in30 cases of gastric cancer,lymph node and paracancerous tissues. The mRNA and protein expression of SATB1 were detected by RTPCR and Western blot in gastric cancer cell line MGC-803 and normal gastric mucosal immortalized cell GES-1. Eukaryotic expression plasmid pc DNA3. 1-His-SATB1( His-SATB1),vector plasmid pc DNA3. 1( His-N),silencing plasmid p PLK+Puro-SATB1 shRNA( shSATB1),silencing vector plasmid p PLK+Puro( sh-N) were transfected into MGC-803 cells using electroporation technology,and after24 h Western blot was used to determine the expression of SATB1,AKT and p-AKT in each cell group. Cell proliferation was detected by CCK8. Results: The expression of SATB1 in gastric cancer tissues,metastatic lymph node tissues and MGC-803 cells significantly increased( P< 0. 05). The protein expression of SATB1 remarkably decreased in sh-SATB1 group,with decreased cell proliferation activity,AKT and p-AKT protein expression( P<0. 05). The protein expression of SATB1 in His-SATB1 group significantly increased,with enhanced cell proliferation,increased AKT and p-AKT protein expression( P<0. 05). Conclusion: SATB1 is highly expressed in gastric cancer tissues and cells. SATB1 promotes the proliferation of gastric cancer cells,which could be achieved by regulating AKT synthesis and activating PI3 K/AKT signaling pathway.
引文
[1]彭智. GLOBCAN胃癌流行病学数据解读[J].肿瘤综合治疗电子杂志,2018,4(4):63-65.Peng Z. Interpretation epidemiological of gastric cancer published by GLOBCAN[J]. J Cancer Comprehensive Therapy,2018,4(4):63-65.
[2] Bray F,Ferlay J,Soerjomataram I,et al. Global cancer statistics2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin,2018,68(6):394-424.
[3] Wang KW,Wang YY,Ye ZY. Expression of Oct 4 in gastric carcinoma and its relationship with invasion,metastasis and prognosis of gastric cancer[J]. Zhejiang Med,2013,35(3):171-174.
[4] Nakayama Y,Mian IS,Kohwi-Shigematsu T. A nuclear targeting determinant for SATB1,a genome organizer in the T cell lineage[J]. Cell Cycle,2005,4(8):1099-1106.
[5] Dickinson LA,Joh T,Kohwi Y,et al. A tissue-specific MAR/SAR DNA-binding protein with unusual binding site recogonition[J].Cell,1992,70(4):631-645.
[6] Han HJ,Russo J,Kohwi Y,et al. SATB1 reprogrammes gene expression to promote breast tumour growth and metastasis[J].Nature,2008,452(7148):187-193.
[7] Huang B,Zhou HL,Wang X,et al. Silencing SATB1 with siRNA inhibits the proliferation and invasion of small cell lung cancer cells[J]. Cancer Cell Int,2013,13(1):8.
[8] Song GQ,Liu K,Bai YG. The expression of SATB1 in SGC-7901cells[J]. Chin J General Pract,2014,1(12):21-23.
[9]袁春銮,孙志超,张磊,等. SATB1阳性患者胃癌临床病理特征分析及预后[J].实用临床医药杂志,2016,20(3):52-56.Yuan CL,Sun ZC,Zhang L,et al. Clinicopathological characteristic and prognosis of patients with SATB1-positive gastric cancer[J].Clin Med J,2016,20(3):52-56.
[10]赵冬梅,耿旭景,谭丽,等. IL-6在子宫内膜异位患者血清中的表达及其潜在作用机制研究[J].中国免疫学杂志,2018,34(11):1707-1715.Zhao DM,Geng XJ,Tan L,et al. Expression of IL-6 in serum of patients with endometriosis and its potential mechanism[J]. Chin J Immunol,2018,34(11):1707-1715.
[11] Notani D,Ramanujam PL,Kumar PP,et al. N-terminal PDZ-like domain of chromatin organizer SATB1 contributes towards its function as transcription regulator[J]. J Biosci,2011,36(3):461-469.
[12] Olasz J,Doleschall Z,Dunai Z,et al. PI3K/AKT pathway activation and therapeutic consequences in breast cancer[J].Magy Onkol,2015,59(4):346-351.
[13] Franke TF,Kaplan DR,Cantley LC. PI3K:downstream AKTion blocks apoptosis[J]. Cell,1997,88(4):435-437.
[14] Burgering BM,Coffer PJ. Protein kinase B(c-AKT)in phosphatidylinositol-3-OH kinase signal transduction[J]. Nature,1995,376(6541):599-602.
[15] Franke TF,Yang SI,Chan TO,et al. The protein kinase encoded by the AKT proto-oncogene is a target of the PDGF-activated phosphatidylinositol 3-kinase[J]. Cell,1995,81(5):727-736.
[16] Tu W,Luo M,Wang Z,et al. Upregulation of SATB1 promotes tumor growth and metastasis in liver cancer[J]. Liver Int,2012,32(7):1064-1078.
[17] Sharma N,Nanta R,Sharma J,et al. PI3K/AKT/m TOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth[J]. Oncotarget,2015,6(31):32039-32060.
[18] Zhong XS,Zheng JZ,Reed E,et al. SU5416 inhibited VEGF and HIF-1alpha expression through the PI3K/AKT/p70S6K1signaling pathway[J]. Biochem Biophy Res Commun,2004,324(2):471-480.
[19]解友邦,李建平,冯建明,等. P13K/AKT信号转导通路和肿瘤[J].国际呼吸杂志,2015,35(19):1502-1505.Xie YB,Li JP,Feng JM,et al. P13K/AKT signal transduction pathway and tumor[J]. Int J Respir,2015,35(19):1502-1505.
[2] Bray F,Ferlay J,Soerjomataram I,et al. Global cancer statistics2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin,2018,68(6):394-424.
[3] Wang KW,Wang YY,Ye ZY. Expression of Oct 4 in gastric carcinoma and its relationship with invasion,metastasis and prognosis of gastric cancer[J]. Zhejiang Med,2013,35(3):171-174.
[4] Nakayama Y,Mian IS,Kohwi-Shigematsu T. A nuclear targeting determinant for SATB1,a genome organizer in the T cell lineage[J]. Cell Cycle,2005,4(8):1099-1106.
[5] Dickinson LA,Joh T,Kohwi Y,et al. A tissue-specific MAR/SAR DNA-binding protein with unusual binding site recogonition[J].Cell,1992,70(4):631-645.
[6] Han HJ,Russo J,Kohwi Y,et al. SATB1 reprogrammes gene expression to promote breast tumour growth and metastasis[J].Nature,2008,452(7148):187-193.
[7] Huang B,Zhou HL,Wang X,et al. Silencing SATB1 with siRNA inhibits the proliferation and invasion of small cell lung cancer cells[J]. Cancer Cell Int,2013,13(1):8.
[8] Song GQ,Liu K,Bai YG. The expression of SATB1 in SGC-7901cells[J]. Chin J General Pract,2014,1(12):21-23.
[9]袁春銮,孙志超,张磊,等. SATB1阳性患者胃癌临床病理特征分析及预后[J].实用临床医药杂志,2016,20(3):52-56.Yuan CL,Sun ZC,Zhang L,et al. Clinicopathological characteristic and prognosis of patients with SATB1-positive gastric cancer[J].Clin Med J,2016,20(3):52-56.
[10]赵冬梅,耿旭景,谭丽,等. IL-6在子宫内膜异位患者血清中的表达及其潜在作用机制研究[J].中国免疫学杂志,2018,34(11):1707-1715.Zhao DM,Geng XJ,Tan L,et al. Expression of IL-6 in serum of patients with endometriosis and its potential mechanism[J]. Chin J Immunol,2018,34(11):1707-1715.
[11] Notani D,Ramanujam PL,Kumar PP,et al. N-terminal PDZ-like domain of chromatin organizer SATB1 contributes towards its function as transcription regulator[J]. J Biosci,2011,36(3):461-469.
[12] Olasz J,Doleschall Z,Dunai Z,et al. PI3K/AKT pathway activation and therapeutic consequences in breast cancer[J].Magy Onkol,2015,59(4):346-351.
[13] Franke TF,Kaplan DR,Cantley LC. PI3K:downstream AKTion blocks apoptosis[J]. Cell,1997,88(4):435-437.
[14] Burgering BM,Coffer PJ. Protein kinase B(c-AKT)in phosphatidylinositol-3-OH kinase signal transduction[J]. Nature,1995,376(6541):599-602.
[15] Franke TF,Yang SI,Chan TO,et al. The protein kinase encoded by the AKT proto-oncogene is a target of the PDGF-activated phosphatidylinositol 3-kinase[J]. Cell,1995,81(5):727-736.
[16] Tu W,Luo M,Wang Z,et al. Upregulation of SATB1 promotes tumor growth and metastasis in liver cancer[J]. Liver Int,2012,32(7):1064-1078.
[17] Sharma N,Nanta R,Sharma J,et al. PI3K/AKT/m TOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth[J]. Oncotarget,2015,6(31):32039-32060.
[18] Zhong XS,Zheng JZ,Reed E,et al. SU5416 inhibited VEGF and HIF-1alpha expression through the PI3K/AKT/p70S6K1signaling pathway[J]. Biochem Biophy Res Commun,2004,324(2):471-480.
[19]解友邦,李建平,冯建明,等. P13K/AKT信号转导通路和肿瘤[J].国际呼吸杂志,2015,35(19):1502-1505.Xie YB,Li JP,Feng JM,et al. P13K/AKT signal transduction pathway and tumor[J]. Int J Respir,2015,35(19):1502-1505.