miR-125b在卵巢癌细胞中的表达及其意义

Expression and Significance of miR-125b in Ovarian Cancer Cells

[目的]探讨miR-125b在卵巢癌细胞中的表达及其意义。[方法]采用实时荧光定量多聚核苷酸链式反应(Real time-PCR)检测miR-125b在人正常卵果上皮细胞和卵巢癌细胞中的表达水平;CCK-8法检测卵巢癌细胞增殖情况;采用°H标记的2-脱氧葡萄糖(^(3)H2-deoxyglucose,^(3)H-2DG)法和乳酸定量试剂盒分别检测卵巢癌细胞葡萄糖摄取和乳酸产生的情况;采用Western blot检测miR-125b mimics过表达后卵巢癌细胞中已糖激酶2(Hexokinase 2,HK2)的表达。[结果]Real time-PCR结果显示:miR-125b在卵巢癌细胞中表达水平显著低于正常卵巢上皮细胞(P<0.05);CCK-8结果显示:miR-125b-mimics转染卵巢癌细胞48 h和72 h后的增殖能力均较对照组明显降低(P<0.05);^(3)H-2DG和乳酸定量结果显示:卵巢癌细胞过表达miR-125b后,葡萄糖摄取量和乳酸产生量明显减少(P<0.05),有氧糖酵解显著抑制;Western blot结果显示:卵巢癌细胞过表达miR 125b后,HK2蛋白表达水平显著降低(P<0.05)。[结论]miR 125b在卵巢癌细胞中低表达,可能通过下调HK2表达抑制卵巢癌细胞的能量代谢和增殖。

【Objective】To investigate the effect and mechanism of miR-125b on energy metabolism and proliferation in ovarian cancer cells.【Methods】Real-time PCR was used to detect the expression level of miR-125b in human ovarian epithelial cells and ovarian cancer cells.The proliferation of ovarian cancer cells was recorded by CCK-8.Glucose uptake and lactic acid production in ovarian cancer cells were measured by ^(3)H 1a-beled 2-deoxyglucose(3H-2DG)method and lactic acid quantitative kit.Western blot was performed for the expression of hexokinase 2(HK2)in ovarian cancer cells after miR-125b-mimics overexpression.[Results]The real time PCR results showed that the expression level of miR-125b in ovarian cancer cells was significantly lower than that in normal ovarian epithelial cells(P<0.05).The CCK-8 results demonstrated that the proliferation capacity of ovarian cancer cells was significantly reduced after transfected with miR-125b-mimics for 48h and 72h compared with the control group(P<0.05).The quantitative results of ^(3)H-2DG and lactic acid showed that glucose intake and lactic acid production were significantly reduced after overexpression of miR-125b in ovarian cancer cells(P<0.05);and aerobic glycolysis was significantly inhibited.Western blot results showed that the expression level of HK2 protein decreased after overexpression of miR-125b in ovarian cancer cells(P<0.05).[Conclusion]The low expression of miR-125b in ovarian cancer cells inhibits the energy metabolism and proliferation of ovarian cancer cells by down-regulating the expression of HK2.