microRNA-145表达对宫颈癌Hela细胞增殖及凋亡的影响

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(P0.05)。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(P0.05)。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(P0.05)。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(P0.05)。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。

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

ABSTRACT 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 of microRNA-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 intensity values(OD490) value of 490nm wave in microRNA-145 group were significantly lower than that of transfection of 0 h; after transfection 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.