目的 探讨微RNA-29b（miR-29b）对子宫内膜癌细胞增殖、迁移和侵袭的影响。方法 子宫内膜癌HEC-1-B细胞分为miR-29b模拟物组（MM组）、miR-29b阻遏物组（MR组）和阴性对照物组（MNC组）,分别转染miR-29b拟似物、miR-29b阻遏物和miR-29b阴性对照物,每组设置6个复孔。以实时定量逆转录PCR检测miR-29b表达,以水溶性四氮唑（WST-1）检测miR-29b对HEC-1-B子宫内膜癌细胞增殖的影响,以Transwell小室检测HEC-1-B子宫内膜癌细胞迁移和侵袭的影响,以Western blot法检测磷酸酶张力蛋白同源物（PTEN）-蛋白激酶 B（AKT）通路蛋白表达水平。结果 MNC组、MM组、MR组miR-29b相对表达量分别为（2 032.1±873.4）、（19 272.8±2 087.9）、（472.7±105.6）,组间比较差异有统计学意义（P<0.05）。MM组0、3、5、7 d时OD值分别为（0.32±0.06）、（0.53±0.08）、（1.13±0.12）和（1.92±0.14）,MNC组0、3、5、7 d时OD值分别为（0.34±0.09）、（0.71±0.08）、（1.67±0.21）和（3.49±0.24）,MR组0、3、5、7 d时OD值分别为（0.38±0.09）、（0.84±0.18）、（2.43±0.24）和（5.67±0.15）,3组0 d时OD值比较差异无统计学意义（P=0.216）,三组3 d、5 d、7 d时OD值比较差异存在统计学意义（P<0.001）。MNC组、MM组和MR组迁移细胞数分别为（403.9±23.8）（102.6±15.7）和（685.7±46.8）个,上述3组侵袭细胞数分别为（82.1±12.7）（38.2±10.6）和（124.6±21.6）个,MM组和MNC组上述指标比较差异均有统计学意义（P<0.05）,MR组和MM组上述指标比较差异均有统计学意义（P<0.05）。MNC组、MM组、MR组PTEN蛋白相对表达量分别为（0.25±0.08）、（0.69±0.11）、（0.11±0.05）,上述3组p-AKT蛋白相对表达量分别为（0.58±0.10）、（0.13±0.06）和（0.79±0.08）,上述3组AKT蛋白相对表达量分别为（0.38±0.09）、（0.37±0.11）和（0.37±0.08）,MM组与MNC组PTEN、p-AKT水平比较差异有统计学意义（P<0.05）,AKT水平比较差异无统计学意义（P>0.05）;MR组与MNC组、MM组PTEN、p-AKT水平比较差异有统计学意义（P<0.05）,AKT水平比较差异无统计学意义（P>0.05）。结论 过表达miR-29b对子宫内膜癌细胞增殖、迁移和侵袭具有抑制作用,靶向PTEN-AKT可能是其重要作用途径。

Objective To investigate the effects of microRNA-29b on proliferation,migration and invasion of endometrial cancer cells．Methods The endometrial cancer HEC-1-B cells were divided into micro29b mimetic group（MM group）,micro29b repressor group（MR group）and negative control group（MNC group）,and the micro29b mimetic,micro29b repressor and micro29b negative control were transfected into each group,six compound holes with each group．The real-time quantitative reverse transcription polymerase chain reaction（RT-PCR）was used to detect the expression of mi29b,WST-1 was used to detect the effect of mi29b on the proliferation of HEC-1-B endometrial cancer cells,Transwell chamber was used to detect the migration and invasion of HEC-1-B endometrial cancer cells,and Western blot was used to detect the expression level of PTEN-AKT pathway protein．Results The relative expression levels of microRNA-29b in MNC group,MM group and MR group were（2 032.1±873.4）,（19 272.8±2 087.9）and（472.7±105.6）,respectively,and there were significant differences between groups（P<0.05）．OD values of MM group at 0 d,3 d,5 d and 7 d were（0.32±0.06）,（0.53±0.08）,（1.13±0.12）and（1.92±0.14）respectively．The OD values of MNC group at 0,3,5 and 7 days were（0.34±0.09）,（0.71±0.08）,（1.67±0.21）and（3.49±0.24）respectively．The OD values of MR group at 0 d,3 d,5 d and 7 d were（0.38±0.09）,（0.84±0.18）,（2.43±0.24）and（5.67±0.15）respectively．There was no significant difference in OD value between the three groups on day 0 （P=0.216）．There were significant differences in OD value between the three groups on day 3,day 5 and day 7（P<0.001）．The number of migrating cells in MNC group,MM group and MR group were（403.9±23.8）cells,（102.6±15.7）cells and（685.7±46.8）cells,respectively．The number of invasive cells in the above three groups were（82.1±12.7）cells,（38.2±10.6）cells and（124.6±21.6）cells．There were significant differences in the above indexes between MM group and MNC group（P<0.05）,also between MR group and MM group（P<0.05）．The relative expression levels of PTEN protein in MNC group,MM group and MR group were（0.25±0.08）,（0.69±0.11）and（0.11±0.05）．The relative expression levels of p-AKT protein in the above three groups were（0.58±0.10）,（0.13±0.06）and（0.79±0.08）．The relative expression levels of AKT protein in the above three groups were（0.38±0.09）,（0.37±0.11）and（0.37±0.08）,respectively．Compared with MNC group,the levels of PTEN and p-AKT in MM group had statistical significance（P<0.05）,but there was no statistical difference in AKT level（P>0.05）．Compared with MNC group and MM group,the levels of PTEN and p-AKT in MR group had statistical significance（P<0.05）,and there was no statistical difference in AKT level（P>0.05）．Conclusions Overexpression of microRNA-29b can inhibit the proliferation,migration and invasion of endometrial cancer cells,and targeting PTEN-AKT may be an important pathway．

目的 研究过表达miR-29a-3p对IL-22诱导的HaCaT细胞增殖的影响。方法 将HaCaT细胞分为Cell组、IL-22组、IL-22+NC组和IL-22+ miR-29a-3p组,荧光定量PCR检测miR-29a-3p的表达水平,CCK8检测细胞的活力,流式细胞仪检测细胞凋亡及周期。结果 与0 μg/L组相比,25 μg/L组、50 μg/L组和100 μg/L组HaCaT细胞的增殖率在24 h、48 h和72 h均出现升高(F值分别为33.27、36.19、52.29,均P<0.000 1)。与0 μg/L组相比,miR-29a-3p在50 μg/L组和100 μg/L组HaCaT中的表达水平降低(F=129,P<0.000 1),分别降低83%和80%。与IL-22+NC组相比,IL-22+ miR-29a-3p组的增殖率在24 h、48 h和72 h均降低(P值分别为0.002 1、0.001 6、0.023 1),细胞总凋亡率增加(6.67±1.06 vs 30.55±1.86,P=0.000 1),G1期细胞比例增加(P=0.000 1),S期细胞比例降低(P=0.000 1)。结论 IL-22可降低HaCaT中miR-29a-3p的表达量,过表达miR-29a-3p通过促进凋亡和引起细胞G1期阻滞抑制IL-22诱发的HaCaT细胞过度增生。

Objective To investigate the effect of miR-29a-3p overexpression on IL-22-induced proliferation of HaCaT cells. Methods HaCaT cells were divided into four groups, Cell group, IL-22 group, IL-22 +NC group and IL-22+miR-29a-3p group. The expression level of miR-29a-3p was detected by fluorescence quantitative PCR. Cell viability was detected by CCK8. Apoptosis and cell cycle were detected by flow cytometry. Results Compared with the 0 g/L group, the proliferation rate of HaCaT cells in the 25 μg/L group, 50 μg/L group and 100 μg/L group was increased at 24 h, 48 h and 72 h (F value was 33.27, 36.19, 52.29,respectively, all P<0.000 1). Compared with the 0 μg/L group, miR-29a-3p expression level in HaCaT in 50 μg/L and 100 μg/L groups was decreased (F=129, P<0.000 1), with a decrease of 83% and 80%, respectively. Compared with the IL-22+NC group, proliferation rate of IL-22+miR-29a-3p group was decreased at 24 h, 48 h and 72 h (P value was 0.002 1, 0.001 6, 0.023 1, respectively), total apoptosis rate was increased (6.67±1.06 vs 30.55±1.86, P=0.000 1), cell proportion in G1 phase was increased (P=0.000 1), and the cell proportion in S phase was decreased (P=0.000 1). Conclusion Il-22 can reduce miR-29a-3p expression level in HaCaT, and miR-29a-3p overexpression can inhibit the excessive proliferation induced by IL-22 in HaCaT cells by promoting apoptosis and inducing G1 phase arrest.