miR-381在肾癌细胞中的表达变化及其对细胞增殖、迁移、侵袭能力的影响

Expression of miR-381 and its effect on proliferation,migration,and invasion of renal carcinoma cells

目的观察miR-381在肾细胞癌(肾癌)细胞系中的表达变化,及其对肾癌细胞增殖、迁移、侵袭能力的影响。方法采用荧光定量PCR法检测肾癌细胞系Caki-2、A498、786-O及人正常肾上皮细胞系HK-2中的miR-381。将Caki-2细胞分为miR-381组、NC组、mock组,miR-381组转染miR-381 mimics,mock组转染miR-381 Scramble,NC组不做转染处理。采用CCK-8试剂盒检测各组细胞增殖能力;采用细胞划痕实验检测各组细胞迁移能力;Transwell实验检测细胞侵袭能力;采用qRT-PCR法检测各组细胞中的DNA结合抑制因子1(ID-1)mRNA,采用Western blotting法检测ID-1蛋白。结果 Caki-2、A498、786-O细胞中miR-381相对表达量低于HK-2细胞(P<0.01)。转染72、96 h后miR-381组细胞增殖能力低于NC组、mock组(P均<0.05)。miR-381组、mock组、NC组细胞划痕愈合率分别为28.7%±2.6%、59.7%±4.8%、56.4%±4.5%(P<0.01),侵袭细胞数分别为(44.7±3.9)、(235.9±17.9)、(225.7±15.7)个,miR-381组细胞划痕愈合率及侵袭细胞数均少于mock组、NC组(P均<0.01)。miR-381组细胞中ID-1 mRNA及蛋白相对表达量均明显低于mock组、NC组(P均<0.01)。结论 miR-381在肾癌细胞中低表达;过表达miR-381可降低肾癌细胞的增殖、迁移、侵袭能力,其机制可能与ID-1表达下调有关。

Objective To observe the expression changes of miR-381 in the renal cell carcinoma cell line,and its effect on the proliferation,migration,and invasion of renal cell carcinoma cells. Methods The expression of miR-381 in three renal cell carcinoma cell lines（ Caki-2,A498,and 786-O） and normal kidney cell line HK-2 was detected by fluorescent quantitative PCR. The Caki-2 cell line was divided into three groups,miR-381 group which was transfect with miR-381 mimics,mock group which ws transfected with miR-381,and the NC group without transfection. CCK-8,Scratch test,and Transwell assay were used to determine the proliferation,migration,and invasion capabilities. The real-time PCR and Western blotting were used to detect the mRNA and protein levels of ID-1 in these three groups. Results The expression level of miR-381 in the Caki-2,A498 and 786-O cells was significantly lower than that in the HK-2 cells（ P 〈 0. 01）. At72 and 96 h after transfection,the proliferation of miR-381 group was lower than that of the NC group and mock group（ P 0. 01）. The wound healing rate was 28. 7 ± 2. 6% in the miR-381 group,56. 4 ± 4. 5% in the NC group,and 59. 7 ±4. 8% in the mock group,and the wound healing rate of the miR-381 group was significantly lower than that in the NC group and mock group（ all P 〈 0. 01）. The invasive cells were 44. 7 ± 3. 9 in the miR-381 group,225. 7 ± 15. 7 in the NC group,and 235. 9 ± 17. 9 in the mock group,and the invasive cells were significantly less than those of the NC group and mock group（ all P 〈 0. 01）. Compared with the NC and mock groups,the mRNA and protein expression of ID-1 in the cells with over-expression of miR-381 was lower（ P 〈 0. 01）. Conclusions The miR-381 is low expressed in the renal cell carcinoma cells. Over-expression of miR-381 can inhibit the proliferation,migration,and invasion of renal cell carcinoma cells,which may be associated with the down-regulated expression of ID-1.