微小RNA-7抑制人卵巢癌细胞株增殖及侵袭能力的研究

目的:探讨微小RNA-7(miR-7)对低转移人卵巢癌细胞株HO-8910、高转移人卵巢癌细胞株HO-8910pm增殖及侵袭转移能力的影响。方法:检测两种细胞株的miR-7表达情况,构建miR-7质粒,通过lipofectmin 2000瞬时转染miR-7低表达的细胞株,通过实时PCR检测转染前后细胞株miR-7以及EGFR mRNA表达情况,Western blot法检测EGFR蛋白表达。细胞划痕实验、transwell实验检测细胞体外迁移及侵袭能力;CCK-8检测增殖能力变化。结果:HO-8910细胞miR-7表达为HO-8910pm的(2.517±0.508)倍;转染后HO-8910pm的miR-7表达提高了(8.015±0.1805)倍,同时抑制EGFRmRNA及蛋白表达,抑制侵袭、迁移、增殖能力(P<0.05);而miR-7表达相对高的HO-8910体外迁移、侵袭及增殖能力均低于HO-8910pm。结论:miR-7可通过调控EGFR的表达抑制HO-8910、HO-8910pm的增殖及侵袭转移能力,miR-7可能为临床治疗卵巢癌提供新靶标。

The effects of miRNA-7 on the invasion and proliferation of human ovarian cancer cell line HO-8910 and HO-8910pm cells

Objective:To explore the effects of miRNA(miR)-7 on the invasion and metastasis and proliferation of human ovarian cancer cell line HO-8910 and HO-8910m in vitro.Methods:MiR-7 plasmid was transiently transferred into cells which expressed low level of miR-7 using lipofectmin 2000 in vitro,then miR-7 expression in HO-8910pm cells was determined using Taqman probe of real time PCR before or after transfection.The expression of EGFR was determined by western blot.The ability of invasion was determined using transwell and migration rate was measured by wound healing assay.The proliferation of cells was accessed using CCK-8 assay.Results:The expression level of miR-7 was significantly increased in miR-7-transfected HO-8910pm cells compared with that in HO-8910pm cells.The expression of EGFR was down-regulated(P<0.05).The ability of proliferation and invasion and metastasis of miR-7-transfected HO-8910pm cells was significantly inhibited(P<0.05).Conclusion:miR-7 inhibits the proliferation and invasion of HO-8910 and HO-8910pm by regulating EGFR expression,suggesting that miR-7 might be a novel target for the biological therapy of human ovarian cancer.