雌二醇联合孕激素对卵巢癌细胞凋亡及microRNA-15a表达的影响

Effect of Estradiol Combined with Progestogen on Apoptosis of Ovarian Cancer Cell and Expression of MicroRNA-15a

关键词:  雌二醇  孕激素  卵巢癌  Bcl-2  Bax  microRNA-15a(miR-15a)

Key words:  estradiol  progesterone  ovarian cancer  Bcl-2  Bax  microRNA-15a(miR-15a)

摘要:

目的 探讨雌二醇（E2）联合孕激素（P4）对microRNA-15a （miR-15a）表达的影响。方法 采集手术中切除的卵巢癌组织标本进行细胞培养，分3个处理组：E2组，P4组和雌二醇联合孕激素（E2+P4）组。激素处理后，MTT法检测细胞存活率；流式细胞技术检测细胞凋亡与细胞周期；qRT-PCR法检测Bcl-2，Bax和miR-15a的相对表达量。结果 高浓度（浓度均为10^-4 mol·L^-1）的E2，P4，E2+P4能够降低卵巢癌细胞的存活率，并表现为时间依赖性；低浓度（≤ 10^-8 mol·L^-1） E2能够提高卵巢癌细胞的存活率。与对照组相比，高浓度E2，P4和E2+P4能够增加卵巢癌细胞的凋亡率（P<0.001），并且E2+P4的作用最明显；低浓度E2能够抑制肿瘤细胞的凋亡（P<0.001）。E2，P4和E2+P4对细胞周期的影响没有统计学差异。高浓度的E2，P4和E2+P4能够下调Bcl-2的表达（P<0.05或P<0.001），上调Bax的表达（P<0.001）；但是低浓度E2作用却相反。高浓度的E2+P4能够促进miR-15a的表达（P<0.001）。结论 高浓度的E2，P4和E2+P4能够降低细胞的存活率促进细胞凋亡，下调Bcl-2的表达，上调Bax的表达；低浓度E2的作用则相反；高浓度的E2+P4能够促进miR-15a的表达。

Abstract:

OBJECTIVE To evaluate the effect of estradiol(E2) combined with progestogen(P4) on expression of microRNA-15a(miR-15a). METHODS The primary ovarian cancer cells from clinical ovarian cancer tissues were isolated and cultured, and divided into 3 groups:E2 group, P4 group, E2 combined with P4(E2+P4) group. The survival rate of ovarian cancer cells after the treatment was analyzed by MTT assay. Apoptosis rate and cell cycle were measured by flow cytometry. Moreover, the relative abundance of miR-15a, Bcl-2 and Bax expressions were detected by qRT-PCR. RESULTS High concentration (10^-4 mol·L^-1) of E2, P4 and E2+P4 inhibited the survival rate of ovarian cancer cells with a time dependent manner. However, low concentration(≤ 10^-8 mol·L^-1) of E2 induced the survival rate of ovarian cancer cells. Compared with control group, high concentration of E2, P4 and E2+P4 induced apoptosis of ovarian cancer cells(P<0.001), and the most obvious effect of promoting apoptosis was E2+P4. However, low concentration of E2 reduced apoptosis of ovarian cancer cells(P<0.001). E2, P4 and E2+P4 had no effect on cell cycle. High concentration of E2, P4 and E2+P4 reduced the expression of Bcl-2(P<0.05 or P<0.001), but they induced the expression of Bax(P<0.001). However, the effect of low concentration of E2 was opposite. Moreover, high concentration of E2+P4 could promote the expression of miR-15a(P<0.001). CONCLUSION High concentration of E2, P4 and E2+P4 significantly inhibit the survival and promot the apoptosis, reduce the expression of Bcl-2, and induce the expression of Bax in ovarian cancer cells. However, the effect of low concentration of E2 is opposite. High concentration of E2+P4 can induce the expression of miR-15a.