微 RNA-125、信号转导与转录活化因子 3对卵巢癌细胞的影响及靶向关系验证

目的检测微 RNA(miR)-125a、信号转导与转录活化因子 3(STAT3)在卵巢癌细胞中的表达水平，分析其对卵巢癌细胞增殖、侵袭和凋亡的影响，并验证两者的靶向关系。方法 2020年 1月至 2021年 12月，采用 qRT-PCR检测卵巢癌细胞和正常人卵巢上皮细胞中 miR-125a、STAT3 mRNA水平。利用转染技术将 SKOV3细胞分为 miR-NC(miR-125a阴性对照组质粒)组、 miR-125a(miR-125a过表达质粒)组、 si-NC(沉默 STAT3阴性对照质粒)组、 si-STAT3(沉默 STAT3质粒)组、 miR-125a+si-NC和 miR-125a+si-STAT3组。 MTT实验、 Transwell实验和凋亡实验分别检测不同组间细胞的 OD值、穿膜细胞数和凋亡率。萤光素酶报告基因分析 SKOV3细胞中 miR-125a与 STAT3的靶向关系，蛋白质印迹法检测不同组间 STAT3蛋白表达情况。结果与 HOSEpiC细胞 miR-125a和 STAT3 mRNA水平( 1.00±0.15、0.54±0.08)相比， SKOV3、CAOV3和 PEO1细胞中 miR-125a水平(0.41±0.07、0.56±0.08、0.58±0.10)降低， STAT3 mRNA水平( 1.87±0.22、1.54±0.07、1.54±0.08)增加；选用 SKOV3细胞进行后续实验。与 miR-NC组相比， miR-125a组细胞增殖能力减弱、穿膜细胞数降低、凋亡率增加。与 si-NC组相比， si-STAT3组细胞增殖能力减弱、穿膜细胞数降低、凋亡率增加。与 miR-125a+si-NC组相比， miR-125a+si-STAT3组细胞增殖能力减弱、穿膜细胞数降低、凋亡率增加。 STAT3+miR-125a mimic组与 STAT3 WT组相比， STAT3蛋白表达较弱，且 miR-125a与 STAT存在靶向结合位点。结论卵巢癌细胞中 miR-125a的过表达可靶向 STAT3抑制 SKOV3细胞的增殖和侵袭并诱导细胞凋亡，发挥抗肿瘤活性。

Effects of miR-125 and STAT3 on ovarian cancer cells and verification of their targeting relationship

Objective To detect the expression levels of micrornas (miR)-125a and STAT3 in ovarian cancer cells, analyze their effects on the proliferation, invasion and apoptosis of ovarian cancer cells, and verify the targeting relationship between miR-125a and STAT3.Methods From January 2020 to December 2021, qRT-PCR was used to detect the relative expression levels of miR-125a and STAT3 mRNA in ovarian cancer cells and normal human ovarian epithelial cells. SKOV3 cells were divided into miR-NC (miR-125a negative control plasmid) group, miR-125a (miR-125a overexpressed plasmid) group, si-NC (silenced STAT3 negative control plasmid) group, si-STAT3 (silenced STAT3 plasmid) group, and miR-125a+si (transfected with si-NC and miR-125a+si-STAT3) groups. MTT experiment, Transwell experiment and apoptosis experiment were used to detect the OD value, number of transmembrane cells and apoptosis rate of SKOV3 cells in different groups. Western blotting was used to detect the expression of STAT3 protein among differentgroups. Luciferase reporter gene was used to analyze the targeting relationship between miR-125a and STAT3 in SKOV3 cells. Re? sults Compared with the mRNA levels of miR-125a and STAT3 in HOSEpiC cells (1.00±0.15, 0.54±0.08), the levels of miR-125a in SKOV3, CAOV3 and PEO1 cells (0.41±0.07, 0.56±0.08, 0.58±0.10) were decreased. STAT3 mRNA levels (1.87±0.22, 1.54±0.07,1.54±0.08) were increased, and SKOV3 cells were selected for follow-up experiments. SKOV3 cells was selected for follow-up experiments. Compared with the miR-NC group, the miR-125a group had weakened cell proliferation, decreased number of transmembranecells, and increased apoptosis rate. Compared with the si-NC group, the cell proliferation ability of the si-STAT3 group was weakened, the number of penetrating cells was reduced, and the apoptosis rate was increased. Compared with the miR-125a+si-NC group, the miR125a+si-STAT3 group had weakened cell proliferation, decreased number of transmembrane cells, and increased apoptosis rate. Compared with the STAT3 WT group, the expression of STAT3 protein in the STAT3+miR-125a mimic group was weaker, and miR-125a had a targeted binding site for STAT.Conclusion Overexpression of miR-125a in ovarian cancer cells can target STAT3 to inhibit theproliferation and invasion of SKOV3 cells and induce cell apoptosis to exert anti-tumor activity.