雌二醇对BG-1卵巢癌细胞增殖及侵袭的影响

目的:探讨雌二醇(E2)对卵巢癌细胞BG-1增殖、侵袭的影响及作用机制。方法:MTT法检测10-9~10-5mol/L E2作用于BG-1细胞72h后的细胞增殖率的变化,并筛选最适作用浓度。将BG-1分为4组:E2组、E2+ICI(ICI182,780 ER阻断剂)组、E2+PPT(特异性ERα激动剂)组和E2+DPN(特异性ERβ激动剂)组。MTT法检测不同药物作用72h后的细胞增殖率;RT-PCR、Western blot法分别检测药物作用6、24h和48h后各组细胞中cyclin(细胞周期素)D1和p21表达水平;ELISA检测10-9~10-5mol/L E2作用后,BG-1细胞中MMP-9水平的变化;Transwell侵袭实验检测E2及ER调节剂作用后BG-1细胞的侵袭力变化。结果:10-9~10-5mol/L E2作用于BG-1细胞72h后,细胞增殖率均高于对照组(P0.05),并且E2浓度为10-7mol/L时BG-1细胞的增殖率最高。不同药物作用后,E2+PPT组的细胞增殖率显著高于E2组(P0.05),E2+ICI组的细胞增殖率显著低于E2组(P0.05),而E2+DPN组细胞增殖率无显著变化(P0.05)。与E2组相比,E2+ICI组中cyclin D1表达明显降低,p21表达显著升高(P0.05);E2+PPT组中cyclin D1与p21的表达水平与E2+ICI组大致相反(P0.05);E2+DPN组中cyclin D1和p21的表达均无明显变化(P0.05)。10-9~10-5mol/L E2均可促进MMP-9分泌(P0.05),其中10-8mol/L E2作用3h后MMP-9分泌最多。E2组的侵袭力显著高于空白对照组,而E2+ICI组的侵袭力显著低于空白对照组(P0.05)。结论:E2通过ERα信号通路促进BG-1细胞增殖;E2还能促进MMP-9分泌,增强BG-1细胞的侵袭力。

The effects of E2on proliferation and invasion in BG-1 ovarian cancer cell

Objective: To investigate the effects of 17β-estradiol( E2) on the proliferation and invasion of BG-1 ovarian cancer cell line and its mechanism. Methods: The proliferation was evaluated by MTT assay after cells were treated with 10-9~ 10-5mol / L E2 to identify the optimal concentration on BG-1. Then cells were divided into 4 groups: E2 group,E2+ ICI group,E2+PPT group and E2+DPN group. The expressions of cyclin D1 and p21 in BG-1 cells were analyzed by PCR and Western blot,after cells were cultured with different drugs for 6,24 and 48h.Matrix metalloproteinase-9( MMP-9) expression was evaluated by ELISA after treated with 10-9~ 10-5mol / L E2 for 3h,6h,12 h and 24 h,respectively. The invasion was detected after cells were treated with E2 and E2+ ICI through transwell invasion assay. Result: After treated with E2,the cell proliferation increased significantly compared with control( P<0.05).Furthermore,the proliferative rate of cells treated with 10-7mol / L E2 was highest among the concentration gradients.The proliferation of E2+PPT group was significantly higher than that of E2group( P<0.05),while that of E2+ICI group was much lower than E2 group. No obvious alteration of proliferation was observed between E2+DPN group and E2group( P >0.05). In E2+ICI group,the expression of cyclinD1 was much lower than E2 group,while the expression of p21 was much higher than E2group( P<0.05).The expressions of cyclinD1 and p21 in E2+PPT group were roughly opposite to E2+ICI group( P<0.05).In E2+DPN group,no significant difference of cyclinD1 and p21 expression was observed compared to E2group( P>0.05).E2 dramatically increased the expression of MMP-9 at concentrations of 10-9~ 10-5mol /L( P < 0. 05). Furthermore,the expression of MMP-9 in cells treated for 3h with 10-8mol /L E2 was much higher than the others.In E2 group,the invasion was enhanced significantly in contrast with blank control group,while in E2+ICI group,it was weaken compared with blank control group( P < 0.05).Conclusions: E2 could promote the proliferation of BG-1 ovarian cancer cell line by up-regulating cyclinD1 and down-regulating p21 through ERα signal pathway. E2 could also stimulate MMP-9 secretion,and enhance the invasion of BG-1 cells.