端粒酶相关因素对培养的视网膜色素上皮细胞增生的影响

目的探讨端粒酶相关因素对培养的视网膜色素上皮（retinal pigment epithelium，RPE）细胞增生的影响。方法用常规培养法培养人RPE细胞后，分别用端粒酶抑制剂二甲基亚砜、热休克蛋白90抑制剂、无血清培养、端粒酶反义寡核苷酸、P16反义寡核苷酸处理，然后提取细胞mRNA，PCR扩增后电泳检测端粒酶mRNA表达。然后用MTT法检测无血清培养组、端粒酶反义寡核苷酸处理组、P16反义寡核苷酸处理组与常规培养组细胞增生状况。结果二甲基亚砜组、热休克蛋白90抑制剂组、无血清培养组、端粒酶反叉寡核苷酸处理组，端粒酶mRNA表达受到抑制，凝胶分析系统软件分析结果电泳H值分别为80、116、90、81，而P16反义寡核苷酸处理组端粒酶mRNA表达增强，电泳H值为255。常规培养组H值为162，阴性对照组为0，阳性对照组（胃癌细胞）为255。MTT法显示：无血清培养组，端粒酶反义寡核苷酸处理组细胞活性下降，细胞生长抑制率上升。而P16反义寡核苷酸处理组细胞活性上升，细胞生长抑制率下降。结论端粒酶抑制剂二甲基亚砜、热休克蛋白90抑制剂、无血清培养、端粒酶反义寡核苷酸可抑制RPE细胞端粒酶mRNA表达，从而抑制RPE细胞增生；P16反义寡核苷酸可促进RPE细胞端粒酶mRNA表达，从而促进RPE细胞增生。

Effect of relative factors of telomerase on the proliferation of cultured retinal pigment epithelial cells

Objective To evaluate the effect of relative factors of telomerase on the proliferation of retinal pigment epithelial(RPE) cells.Methods RPE cells were cultured by normal method and were treated with inhibitor of telomerase-dimethyl sulfoxide(DMSO),inhibitor of heat shock protein 90,free-serum culture,anti-sense oligonucleotide(AON) of telomerase,anti-sense oligonucleotide of P16 respectively.Then the expression of telomerase mRNA was detected by polymerase chain reaction;the proliferation of above groups cells by MTT was detect.Results Expression of telomerase in DMSO group,inhibitor of heat shock protein 90,free-serum culture group,anti-sense oligonucleotide of telomerase group were inhibited,and the electrophororesis H values with gel analysis system was 80,116,90,81,respectively.The expression of telomerase in group of anti-sense oligonucleotide of P16 was enhanced,with the electrophororesis H value of 255.It in normal culture group was 162;0 in negative group and 255 in positive control group (gastric cancer cells).MTT showed that the cytoactive of RPE cells in group of free-serum culture,anti-sense oligonucleotide of telomerase descend and restrain rate of cells ascend,the anti-sense oligonucleotide of P16 group was on the opposite.Conclusion Inhibitor of telomerase DMSO,inhibitor of heat shock protein 90,free-serum culture,anti-sense oligonucleotide of telomerase can inhibit the expression of telomerase in RPE cells,resulting in the proliferation of RPE cells.The anti-sense oligonucleotide of P16 is on the opposite.