铜绿假单胞菌制剂经由甘露糖依赖方式影响人宫颈癌Hela细胞株的生长研究
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摘要
目的研究铜绿假单胞菌制剂(pseudomonas aeruginosa with mannose sensitive hemagglutination pili,PAMSHA)由甘露糖依赖方式影响Bad、Bax及Bcl-2的表达进而影响癌细胞的增殖和细胞周期进程,为治疗宫颈癌的新的生物制剂提供依据。方法采用单独处理PA-MSHA或甘露糖与PA-MSHA联合处理宫颈癌Hela细胞,随后采用MTT法检测细胞增殖,流式检测细胞周期,荧光定量PCR和western blot检测凋亡抑制蛋白Bcl-2和促凋亡蛋白Bad、Bax的表达变化。结果 PA-MSHA单独处理宫颈癌Hela后细胞增殖减慢,细胞周期阻滞在G0/G1期,同时荧光定量PCR和western blot检测显示促凋亡蛋白Bad、Bax的表达增加而凋亡抑制蛋白Bcl-2表达减少。而甘露糖与PA-MSHA共培育组与空白组对比在细胞增殖、细胞周期及凋亡蛋白表达上差异无统计学意义。结论 PA-MSHA可以影响Bad、Bax及Bcl-2的表达进而影响癌细胞的增殖和细胞周期进程,且这种作用为甘露糖敏感型,该结果为其成为治疗宫颈癌的新的生物制剂提供了可靠依据。
Objective To investigate the effects of Pseudomonas aeruginosa with mannose sensitive hemagglutinin( PA- MSHA) on the proliferation and cell cycle of cervical cancer cell line Hela via a mannose- dependent manner. Methods Hela cells were treated with PA- MSHA or PA- MSHA and mannose. The cell proliferation was observed by MTT. Cell cycle distribution was measured by flow cytometry( FCM). Real time RT- PCR and western blot were used to evaluate the expressions of apoptosis- related molecules,Bad,Bax and Bcl- 2. Results For the Hela cells treated with PA- MSHA alone,the cell proliferation slowed down. G0- G1 cell cycle arrest was observed. The expressions of Bad and Bax were strongly increased while that of Bcl- 2 was deceased. For the Hela cells co- cultured with PA- MSHA and mannose,the cell proliferation,cell cycle and the expression of the apoptosis- related proteins were not significantly different from those of the controls( P > 0. 05). Conclusions PA- MSHA can affect the expression of Bad,Bax and Bcl- 2 and further influcence the proliferation and cell cycle of the cancer cells. The effects are mannose- sensitive. The data provide mechanistic details for the potential application of PA- MSHA based treatment against human cervical cancer.
Objective To investigate the effects of Pseudomonas aeruginosa with mannose sensitive hemagglutinin( PA- MSHA) on the proliferation and cell cycle of cervical cancer cell line Hela via a mannose- dependent manner. Methods Hela cells were treated with PA- MSHA or PA- MSHA and mannose. The cell proliferation was observed by MTT. Cell cycle distribution was measured by flow cytometry( FCM). Real time RT- PCR and western blot were used to evaluate the expressions of apoptosis- related molecules,Bad,Bax and Bcl- 2. Results For the Hela cells treated with PA- MSHA alone,the cell proliferation slowed down. G0- G1 cell cycle arrest was observed. The expressions of Bad and Bax were strongly increased while that of Bcl- 2 was deceased. For the Hela cells co- cultured with PA- MSHA and mannose,the cell proliferation,cell cycle and the expression of the apoptosis- related proteins were not significantly different from those of the controls( P > 0. 05). Conclusions PA- MSHA can affect the expression of Bad,Bax and Bcl- 2 and further influcence the proliferation and cell cycle of the cancer cells. The effects are mannose- sensitive. The data provide mechanistic details for the potential application of PA- MSHA based treatment against human cervical cancer.
引文
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[2]李惠芳,常艳丽,李娜,等.P53突变、表达及人乳头状瘤病毒感染与宫颈癌[J].生物化学与生物物理进展,2001,28(2):214-217.
[3]Steben M,Duarte-Franco E.Human papillomavirus infection:epidemiology and pathopgysiology[J].Gynecol Oncol,2007,107(2supp11):S2-S5.
[4]许雪梅.人乳头瘤病毒及宫颈癌疫苗的研究[J].生物化学与生物物理进展,2008,35(10):1095-1103.
[5]李筱梅,郑英,张庆.宫颈癌发病率的年轻化趋势与人乳头瘤病毒感染的关系[J].中国实用妇科与产科杂志,2006,16(9):741-742.
[6]王维,丁王景王聿,王大成.铜绿假单胞菌特有的Tsi2三维结构-一种全新卷曲螺旋构象的类抗毒素蛋白[J].生物化学与生物物理进展,2012,39(7):640-646.
[7]Lee Nq,Jung SB,Ahn BY,et al.Immunization of burn-patients with a Pseudomonas aeruginosa outer membrane protein vaceine elieits antibodies with protective effieaey[J].Vaceine,2000,18(18):1952-1961.
[8]牟希亚.铜绿假单胞菌制剂甘露糖敏感血凝菌毛株菌苗的成功建立[J].微生物学杂志,1986,26:176-179.
[9]Li Z,Hao D,Zhang H,et al.A clinical study on PA-MSHA vaccine used for adjuvant therapy of lymphoma and lung cancer[J].Huaxi Med Coll J Med,2000,31:334-337.
[10]Ling W,Liu H,Cao H,et al.Effects of PA-MSHA vaccine on gastric cancer cells in vitro[J].Chin J Cancer Prev Treat,2008,15:1381-1385.
[11]陈卫东,温继军.TAC方案联合使用铜绿假单胞菌制剂在乳腺癌新辅助化疗中的应用研究[J].南方医科大学学报,2009,29(6):1204-1207.
[12]Jenkins CE,Swiatoniowski A,Issekutz AC,et al.Pseudomonas aeruginosa exotoxin A induces human mast cell apoptosis by a caspase-8 and-3-dependent mechanism[J].J Biol Chem,2004,279:37201-37207.
[13]Jia L,Wang C,Kong H,et al.Effect of PA-MSHA vaccine on plasma phospholipids metabolic profiling and the ratio of Th2/Th1 cells within immune organ of mouse IgA nephropathy[J].J Pharm Biomed Anal,2007,43(2):646-654.
[14]Cao Z,Shi L,Li Y,et al.Pseudomonas aeruginosa:mannose sensitive hemagglutinin inhibits the growth of human hepatocarcinoma cells via mannose-mediated apoptosis[J].Dig Dis Sci,2009,54(10):2118-2127.
[15]Sharon N.Carbohydrates as future anti-adhesion drugs for infectious diseases[J].Biochim Biophys Acta,2006,1760(4):527-537.
[16]King SS,Young DA,Nequin LG,et al.Use of specific sugars to inhibit bacterial adherence to equine endometrium in vitro[J].Am J Vet Res,2000,61(4):446-449.
[17]Eshdat Y,Sharon N.Recognitory bacterial surface lectins which mediate its mannose-specific adherence to eukaryotic cells[J].Biol Cell,1984,51(2):259-266.
[18]Jain RK,Forbes NS.Can engineered bacteria help control cancer[J].Proc Natl Acad Sci USA,2001,98(26):14748-14750.
[19]Anderson JC,Clarke EJ,Arkin AP,et al.Environmentally controlled invasion of cancer cells by engineered bacteria[J].J Mol Biol,2006,355(4):619-627.