慢病毒介导ACTHR过表达对人肾上腺皮质癌H295R细胞的增殖影响

Lentivirus-mediated over-expression of ACTHR on the proliferation of human adrenocortical H295R cells

目的研究ACTHR高表达对人肾上腺皮质癌H295R细胞的影响,并探讨ACTHR在肾上腺肿瘤发病中的作用。方法用慢病毒包装的ACTHR高表达载体感染H295R细胞作为实验组,同时设立对照组。用WST-1法及细胞计数法检测感染后细胞的增殖情况,感染48h后TUNEL法检测细胞的凋亡情况。结果 WST-1增殖检测发现实验组细胞增殖活性呈双重变化趋势,在前3天内增殖活性高于对照组细胞,尤其是第2天达到顶峰,但第4天后增殖活性低于对照组细胞;细胞计数法显示在增殖活性最高时,实验组和对照组细胞分别为(5.3±0.21)×103和(3.8±0.24)×103个,两组细胞计数结果对比具有统计学意义(P<0.05);而TUNEL结果显示荧光镜下两组细胞的凋亡情况无明显差异(P>0.05)。结论 ACTHR对肾上腺肿瘤的影响具有时间依赖性,但可能主要以促进细胞增殖为主,而对凋亡影响不明显。

Objective To explore the effect of adrenocorti cotrophic hormone receptor （ACTHR） over‐expression on the proliferation of human adrenocortical H295R cells ,and to investigate its role in the pathogenesis of adrenal tumors .Methods H295R cells transfected by lentivirus‐mediated ACTHR served as the experimental group .H295R cells without virus infection served as control group .The cell proliferation at different ACTHR levels was evaluated with WST‐1 and cell count .Cell apoptosis after 48 hours was measured with TUNEL .Results Higher proliferation activity was observed in the experimental cells than in the control cells in the first three days ,which peaked on the second day .But this trend reversed on the fourth day . The number of experimental and control cells were （5 .3 ± 0 .21） × 103 and （3 .8 ± 0 .24） × 103 at peak stage ,（ P 〈 0 .05） .By contrast ,there was no significant difference between the two groups in cell apoptosis .Conclusion ACTHR affects H295R cells in a time‐dependent manner .It has stronger effect on the proliferation than on the apoptosis of H295R cells .