摘要
目的:通过检测不同压力CO2气腹环境下结直肠癌细胞周期和microRNA-221(miR-221)及CDKN1C/p57表达的变化,探讨CO2气腹对结直肠癌细胞增殖的影响。方法:将人结直肠癌CaCO2细胞系分别置于0、10、12和15mm Hg的CO2气腹环境下培养4h,应用实时荧光定量PCR检测CaCO2细胞中miR-221表达状况,运用Western-blot分析CDKN1C/p57蛋白表达状况,并通过噻唑蓝(MTT)比色法和流式细胞仪观察细胞的增殖状态和分析细胞周期。结果:与对照组相比,15mm Hg的CO2气腹组结直肠癌细胞中miR-221表达量下降[(1.212±0.159)vs.(1.937±0.208)],而CDKN1C/p57蛋白表达增高[(2.017±0.808)vs.(0.972±0.416)],同时细胞增殖抑制率[(32.7±5.1)%vs.(21.8±3.6)%]增加,且G0/G1期细胞比例明显增加,S期细胞比例显著下降,差异均有统计学意义(P<0.01);0、10、12mm Hg的CO2气腹组结直肠癌细胞增殖抑制率及miR-221、CDKN1C/p57表达水平与对照组相比差异均无统计学意义(P>0.05)。结论:临床常用压力的CO2气腹对结直肠癌细胞增殖无明显影响;15mm Hg的CO2气腹可抑制癌细胞增殖,其原因可能与miR-221表达下降,继而CDKN1C/p57蛋白表达增高有关。
Objective:To investigate the effects of CO2 pneumoperitoneum of different pressures on cell proliferation and expression of microRNA-221(miR-221) and CDKN1C/p57 in colorectal carcinoma cells.Methods:Human colorectal carcinoma cell line CaCO2 were exposed to 0,10,12 and 15mm Hg CO2 pneumoperitoneum in vitro for four hours.The miR-221 expression pattern was detected by real-time RT-PCR following the protocols recommended by commercial corporation.The protein level of CDKN1C/p57 expression was detected by Western-blot analysis.Furthermore,cell proliferation and cell cycle analysis were measured by MTT assay and flow cytometer.Results:The cellular proliferation inhibition rate in 15mm Hg CO2 pneumoperitoneum group enhanced significantly compared with control group [(32.7±5.1)% vs.(21.8±3.6)%],while that in other groups did not changed significantly.The expression of miR-221 was significantly down-regulated in 15mm Hg CO2 pneumoperitoneum group compared with control group [(1.212±0.159) vs.(1.937±0.208)],while CDKN1C/p57 protein expression enhanced markedly [(2.017±0.808) vs.(0.972±0.416)].Conclusions:There was no obvious effects of clinical CO2 pneumoperitoneum on colorectal carcinoma cell proliferation;15 mm Hg CO2 pneumoperitoneum could inhibit cell proliferation by inhibiting miR-221 expression and up-regulating CDKN1C/p57 protein expression consequently.
出处
《腹腔镜外科杂志》
2011年第4期267-271,共5页
Journal of Laparoscopic Surgery
基金
广东省自然科学基金博士启动项目(编号:8451051501000390)
广东省医学科研基金项目(编号:B2010190)
南方医科大学南方医院院长基金项目(编号:2008C005)
关键词
结直肠肿瘤
细胞增殖
CO2气腹
Colorectal neoplasms
Cell proliferation
Carbon dioxide pneumoperitoneum