微小RNA-143抑制人肾癌细胞株A498增殖及其对凋亡的影响

Effect of microRNA-143 on proliferation and apoptosis of human renal cell carcinoma cell line A498

目的观察微小RNA-143（miR-143）对肾细胞癌细胞株A498增殖及凋亡的影响。方法化学合成miR-143模拟物（miR-143mimics）及阴性对照转染对照序列，用Lipofectamine%^TM 2000脂质体瞬时转染miR-143模拟物及阴性对照转染对照序列至肾细胞癌株A498中，miR．143mimics和转染对照序列浓度为60nmol／L，实验分为3组：miR．143mimics转染组、阴性对照转染组（NC组）、空白对照组（Opti—MEMI无血清培养基培养的A498细胞）。采用细胞计数试剂盒（CCK-8）和流式细胞技术等方法检测转染后不同分组肾癌细胞株生长、增殖与凋亡的变化。结果与对照组比较，CCK-8检测结果表明miR-143mimics转染组细胞增殖抑制明显，且抑制效应于转染48h后呈现。流式细胞术检测结果显示miR-143mimics转染组细胞凋亡率增加[（9．10-1-2．31）％]，显著高于阴性对照转染组[（4．23±1．52）％]与空白对照组（（3．43±J．61）％]，组间差异有统计学意义（P〈0．05）。结论miR-143模拟物可抑制肾癌细胞的增殖，诱导肾癌细胞凋亡，具有明显的肿瘤抑制效应。

Objective To observe the effect of microRNA-143 （miR-143） on proliferation and apoptosis of human renal cell carcinoma cell line A498. Methods The miR-143 mimics and negative con- trol transfection sequence were transfected into A498 cells with Lipo2000 liposome, and the concentrations of both re{R-143 mimics and negative control transfection sequence were 60 nmol/L. The experiment was divided into three groups： miR-143 mimics-transfected group, negative control-transfeeted group, blank control group which was cultured with opti-MEMI serum-free culture medium only. After transfection, the cell proliferation and apoptosis were detected by cell counting kit-8 （CCK-8） assay and flow cytometry re- spectively. Results After transfeetion of miR-143 mimics, as compared with control group, CCK-8 assay revealed a significant inhibition of proliferation, and inhibition effect occurred at 48th h after transfeetion. Flow cytometry demonstrated that miR-143 mimics transfeetion increased apotosis rate [ （9. 10±2.31）% ], significantly higher than the negative control group [ （4.23±1.52）%] and blank control group [ （ 3.43±1.61 ） % ] , and there was significant difference among the groups （ P 〈 0. 05 ）. Conclu- sion miR-143 could inhibit proliferation and induce apoptosis of renal cell carcinoma cells, suggesting that miR-143 was a tumor suppressor gene in renal cell carcinoma.