microRNA-145表达对宫颈癌Hela细胞增殖及凋亡的影响

Effect of micro RNA-145 Expression on Proliferation and Apoptosis of Cervical Cancer Hela Cells

目的:探讨微小核糖核酸145(micro RNA-145)表达对宫颈癌Hela细胞增殖及凋亡的影响。方法:实验室常规培养宫颈癌Hela细胞并分为4组,空白(Blank)组(Hela细胞+RPMI1640)、micro RNA-145组(Hela细胞+RPMI1640+micro RNA-145-5p mimics)、阴性序列(NC)组(Hela细胞+RPMI1640+NC)、Mock组(Hela细胞+RPMI1640+Lipofectamine 2000),记录各组Hela细胞转染率,采用实时荧光定量聚合酶链锁反应(QRT-PCR)检测各组Hela细胞中micro RNA-145的表达水平,采用四甲基偶氮唑蓝(MTT)比色法检测Hela细胞增殖情况,采用4',6-二脒基-2-苯基吲哚(DAPI)染色法判断Hela细胞凋亡情况。结果:本研究中,各组Hela细胞转染率均>80%;micro RNA-145组micro RNA-145的表达显著高于Blank组、NC组和Mock组,差异有统计学意义(P<0.05)。转染24 h、48 h、72 h后,micro RNA-145组490 nm波长处的光密度值(OD490值)较转染0h后明显降低,转染48 h、72 h后,Blank组、NC组、Mock组OD490值较转染0 h后时明显升高,转染24 h、48 h、72 h后,micro RNA-145组OD490值均低于Blank组、NC组、Mock组,差异有统计学意义(P<0.05)。DAPI染色后,micro RNA-145组Hela细胞凋亡率高于Blank组、NC组、Mock组,差异有统计学意义(P<0.05)。转染后,Blank组、NC组、Mock组的micro RNA-145表达率、OD490值、DAPI染色后Hela细胞凋亡率比较差异均无统计学意义(P>0.05)。结论:micro RNA-145表达上调可抑制宫颈癌Hela细胞增殖,并促进Hela细胞凋亡,通过药物调控micro RNA-145表达有望成为宫颈癌治疗的新靶点。

Objective： To explore the effect of micro Ribonucleic Acid 145 （microRNA-145） expression on the proliferation and apoptosis of cervical cancer Hela cells. Methods： The cervical cancer Hela cells were cultured routinely in the laboratory and divided into blank group （Hela cell ＋ RPMI1640）, microRNA-145 group （Hela cell ＋ RPMI1640 ＋ microRNA-145-5p mimics）, NC group （Hela cell ＋ RPMI1640 ＋ NC） and Mock group （Hela cell ＋ RPMI1640 ＋ Lipofectamine 2000）. The transfection rate of Hela cells in each group was recorded, the expression of microRNA-145 in Hela cells was detected by real-time quantitative polymerase chain reaction （QRT-PCR）, the proliferation of Hela cells was detected by four methyl blue tetrazolium （MTT） colorimetric assay, the apoptosis of Hela cells was determined by 4＇,6-diamidino-2-phenylindole （DAPI） staining. Results： In this study,the transfection rate of Hela cells in each group was more than 80%; the expression ofmicroRNA-145 in microRNA-145 group was significantly higher than that in blank group, NC group and Mock group, the differences were statistically significant （P〈0.05）. After transfection of 24 h, 48 h and 72 h, the light in- tensity values（OD490） value of 490nm wave in microRNA-145 group were significantly lower than that oftransfection of 0 h; after trans- fection of 48 h and 72 h, the OD490 value of blank group, NC group and Mock group were significantly higher than that of transfection of 0 h; after transfection of 24 h, 48 h and 72 h, the OD490 values in the microRNA-145 group were lower than those in the blank group, the NC group and the Mock group, the differences were statistically significant （P〈0.05）. After DAPI staining, the apoptosis rate of Hela cells in microRNA-145 group was higher than that in blank group, NC group and Mock group, the difference was statistically significant （P〈0.05）. After transfection, there were no significant differences in the microRNA-145 expression rate, OD490 value and Hela cell apoptosis rate after DAPI staining in blank group, NC group and Mock group（P〉0.05）. Conclusion： Up-regulation of microRNA-145 ex- pression can inhibit the proliferation of cervical cancer Hela cells and promote apoptosis of Hela cells,and the regulation of microR- NA-145 expression by drugs is expected to be a new target for cervical cancer treatment.