孕激素受体ser294磷酸化对卵巢癌细胞的增殖和凋亡的影响

发布时间：2018-02-03 15:36

本文关键词： 卵巢癌孕激素受体磷酸化增殖凋亡　出处：《遵义医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨孕激素对卵巢癌细胞孕激素受体Ser294磷酸化状态的影响以及孕激素受体Ser294磷酸化对卵巢癌细胞的增殖和凋亡的影响。方法:(1)将卵巢癌HO8910(PR高表达)和SKOV3(PR低表达)细胞分为四组:R5020组、R5020+U0126组、U0126组和对照组。R5020组于起始时加入不含药物培养基,6h、18h、30h、42h更换为终浓度为10~(-6)mol/L R5020的培养基,12h、24h、36h更换为不含药物的培养基。R5020+U0126组起始加入终浓度为10~(-5)mol/L U0126的培养基,6h、18h、30h、42h更换为终浓度为10~(-6)mol/L R5020的培养基,12h、24h、36h更换为终浓度为10~(-5)mol/L U0126的完全培养基。U0126组加入终浓度为10~(-5)mol/L U0126的培养基。对照组加不含药物的完全培养基,以上四组均于48h收获细胞,应用Western-Blot检测分别各组细胞PRA Ser294和PRB Ser294磷酸化相对表达,由于PR各亚型中仅有PRA Ser294和PRB Ser294会发生磷酸化,所以我们用PRA Ser294和PRB Ser294磷酸化的变化趋势来表示PR Ser294磷酸化的变化趋势。(2)实验分组同上,应用CCK8法和Annexin V/PI双染法流式细胞技术检测各组细胞的增殖抑制率和凋亡率。结果:(1)空白对照组SKOV3和HO8910细胞系PRB Ser294磷酸化相对表达量分别为(0.006±0.002、0.007±0.002);PRA Ser294不发生磷酸化,U0126组SKOV3和HO8910细胞系PRB Ser294磷酸化相对表达量分别为(0.005±0.001、0.006±0.001);PRA Ser294均不发生磷酸化,R5020组SKOV3和HO8910细胞系PRB Ser294磷酸化相对表达量分别为(0.72±0.028、0.68±0.045);PRA Ser294磷酸化相对表达量分别为(0.18±0.027、0.19±0.028),R5020+U0126组SKOV3和HO8910细胞系PRB Ser294磷酸化相对表达量分别为(0.52±0.033、0.37±0.012);HO8910细胞系PRA Ser294磷酸化相对表达量分别为(0.14±0.016);SKOV3细胞PRA Ser294不发生磷酸化。R5020组两株细胞PRA Ser294和PRB Ser294磷酸化相对表达均高于对照组,R5020+U0126组PRA Ser294和PRB Ser294磷酸化相对表达均低于R5020组,差异具有统计学意义(P0.05)。(2)U0126组SKOV3和HO8910细胞系增殖抑制率分别为(9.27%、28.22%),R5020组SKOV3和HO8910细胞系增殖抑制率分别为(40.36%、53.15%),R5020+U0126组SKOV3和HO8910细胞系增殖抑制率分别为(29.65%、38.66%)。R5020组两株细胞的增殖抑制率均高于R5020+U0126组,R5020+U0126组两株细胞的增殖抑制率均高于U0126组,差异具有统计学意义(P0.05)。(3)对照组SKOV3和HO8910细胞系凋亡率分别为(9.79%、9.57%),U0126组SKOV3和HO8910细胞系凋亡率分别为(12.37%、12.62%),R5020组SKOV3和HO8910细胞系凋亡率分别为(21.66%、27.82%),R5020+U0126组SKOV3和HO8910细胞系凋亡分别为(15.35%、14.11%)。R5020组两株细胞凋亡率均高于对照组和R5020+U0126组,R5020+U0126组凋亡率均高于U0126组,差异具有统计学意义(P0.05)。结论:孕激素可使卵巢癌HO8910和SKOV3细胞中PR Ser294磷酸化相对表达增加。卵巢癌SKOV3和HO8910细胞系PR Ser294磷酸化增加后,孕激素抑制卵巢癌细胞系增殖和促凋亡作用增强。
[Abstract]:Objective: to investigate the effect of progesterone on the phosphorylation of progesterone receptor Ser294 in ovarian cancer cells and the effect of progesterone receptor Ser294 phosphorylation on proliferation and apoptosis of ovarian cancer cells. 1) High expression of HO8910(PR and low expression of SKOV3(PR) were divided into four groups: R5020 group. R5020 U0126 group and control group. R5020 group were treated with the drug free medium for 6 h and 18 h for 30 h. After 42h, the medium was replaced with a final concentration of 10 ~ 6 mol / L R5020 for 12 h and 24 h. After 36 h, the medium. R5020 U0126 was replaced with the medium containing no drug. The initial addition of the medium with the final concentration of 10 ~ 5mol / L U0126 was 6 h ~ 18 h ~ (-1) and 30 h ~ (-1). After 42h, the medium was replaced with a final concentration of 10 ~ 6 mol / L R5020 for 12 h and 24 h. After 36 hours, the final concentration was replaced with a complete medium with final concentration of 10g / L mol / L U0126. Group U0126 was added with a final concentration of 10g / L / L U0126. The culture medium of mol/L U0126. The control group was supplemented with complete medium without drug. The above four groups were harvested at 48 h. The relative expression of PRA Ser294 and PRB Ser294 was detected by Western-Blot. Only PRA Ser294 and PRB Ser294 were phosphorylated in the subtypes of PR. So we use the trend of PRA Ser294 and PRB Ser294 phosphorylation to express the change trend of PR Ser294 phosphorylation. CCK8 assay and Annexin V / Pi double staining flow cytometry were used to detect the proliferation inhibition rate and apoptosis rate of the cells in each group. The relative phosphorylation level of PRB Ser294 in SKOV3 and HO8910 cells was 0.006 卤0.002, respectively. 0.007 卤0.002; No phosphorylation occurred in PRA Ser294. The relative phosphorylation levels of PRB Ser294 in U0126 SKOV3 and HO8910 cell lines were 0.005 卤0.001t0. 006 卤0. 001C, respectively. No phosphorylation occurred in PRA Ser294. The relative phosphorylation of PRB Ser294 in R5020 SKOV3 and HO8910 cells was 0.72 卤0.028 and 0.68 卤0.045, respectively. The relative phosphorylation level of PRA Ser294 was 0.18 卤0.027 + 0.19 卤0.028). The relative phosphorylation of PRB Ser294 in R5020 U0126 SKOV3 and HO8910 cell lines was 0.52 卤0.033, respectively. 0.37 卤0.012; The relative phosphorylation level of PRA Ser294 in HO8910 cell line was 0.14 卤0.016; The relative expression of PRA Ser294 and PRB Ser294 phosphorylation in PRA Ser294 of SKOV3 cells was higher than that in control group. The relative expression of PRA Ser294 and PRB Ser294 in R5020 U0126 group was lower than that in R5020 group. The inhibitory rates of proliferation of SKOV3 and HO8910 cell lines in group P0.05 and U0126 were 9.27 and 28.22, respectively. The inhibitory rates of SKOV3 and HO8910 cell lines in R5020 group were 40.36 and 53.15 respectively. The proliferation inhibition rate of SKOV3 and HO8910 cells in R5020 U0126 group was 29.65%. The inhibition rate of proliferation of the two cells in R5020 group was higher than that in R5020 U0126 group. The inhibition rate of proliferation in R5020 U0126 group was higher than that in U0126 group. The apoptosis rates of SKOV3 and HO8910 cell lines in the control group were 9.79 and 9.57, respectively. The apoptosis rate of SKOV3 and HO8910 cells in U0126 group was 12.37 and 12.62 respectively. The apoptosis rate of SKOV3 and HO8910 cells in R5020 group was 21.66 and 27.82 respectively. Apoptosis of SKOV3 and HO8910 cells in R5020 U0126 group was 15.35%. The apoptotic rate in R5020 group was higher than that in control group and R5020 U0126 group, and the apoptosis rate in R5020 U0126 group was higher than that in U0126 group. The difference was statistically significant (P0.05). Conclusion: progesterone can increase the expression of PR Ser294 phosphorylation in HO8910 and SKOV3 cells. The phosphorylation of Ser294 was increased. Progesterone inhibits proliferation and promotes apoptosis of ovarian cancer cell line.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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