本文關(guān)鍵詞： 卵巢癌 孕激素受體 磷酸化 增殖 凋亡　出處：《遵義醫(yī)學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討孕激素對(duì)卵巢癌細(xì)胞孕激素受體Ser294磷酸化狀態(tài)的影響以及孕激素受體Ser294磷酸化對(duì)卵巢癌細(xì)胞的增殖和凋亡的影響。方法:(1)將卵巢癌HO8910(PR高表達(dá))和SKOV3(PR低表達(dá))細(xì)胞分為四組:R5020組、R5020+U0126組、U0126組和對(duì)照組。R5020組于起始時(shí)加入不含藥物培養(yǎng)基,6h、18h、30h、42h更換為終濃度為10~(-6)mol/L R5020的培養(yǎng)基,12h、24h、36h更換為不含藥物的培養(yǎng)基。R5020+U0126組起始加入終濃度為10~(-5)mol/L U0126的培養(yǎng)基,6h、18h、30h、42h更換為終濃度為10~(-6)mol/L R5020的培養(yǎng)基,12h、24h、36h更換為終濃度為10~(-5)mol/L U0126的完全培養(yǎng)基。U0126組加入終濃度為10~(-5)mol/L U0126的培養(yǎng)基。對(duì)照組加不含藥物的完全培養(yǎng)基,以上四組均于48h收獲細(xì)胞,應(yīng)用Western-Blot檢測(cè)分別各組細(xì)胞PRA Ser294和PRB Ser294磷酸化相對(duì)表達(dá),由于PR各亞型中僅有PRA Ser294和PRB Ser294會(huì)發(fā)生磷酸化,所以我們用PRA Ser294和PRB Ser294磷酸化的變化趨勢(shì)來(lái)表示PR Ser294磷酸化的變化趨勢(shì)。(2)實(shí)驗(yàn)分組同上,應(yīng)用CCK8法和Annexin V/PI雙染法流式細(xì)胞技術(shù)檢測(cè)各組細(xì)胞的增殖抑制率和凋亡率。結(jié)果:(1)空白對(duì)照組SKOV3和HO8910細(xì)胞系PRB Ser294磷酸化相對(duì)表達(dá)量分別為(0.006±0.002、0.007±0.002);PRA Ser294不發(fā)生磷酸化,U0126組SKOV3和HO8910細(xì)胞系PRB Ser294磷酸化相對(duì)表達(dá)量分別為(0.005±0.001、0.006±0.001);PRA Ser294均不發(fā)生磷酸化,R5020組SKOV3和HO8910細(xì)胞系PRB Ser294磷酸化相對(duì)表達(dá)量分別為(0.72±0.028、0.68±0.045);PRA Ser294磷酸化相對(duì)表達(dá)量分別為(0.18±0.027、0.19±0.028),R5020+U0126組SKOV3和HO8910細(xì)胞系PRB Ser294磷酸化相對(duì)表達(dá)量分別為(0.52±0.033、0.37±0.012);HO8910細(xì)胞系PRA Ser294磷酸化相對(duì)表達(dá)量分別為(0.14±0.016);SKOV3細(xì)胞PRA Ser294不發(fā)生磷酸化。R5020組兩株細(xì)胞PRA Ser294和PRB Ser294磷酸化相對(duì)表達(dá)均高于對(duì)照組,R5020+U0126組PRA Ser294和PRB Ser294磷酸化相對(duì)表達(dá)均低于R5020組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。(2)U0126組SKOV3和HO8910細(xì)胞系增殖抑制率分別為(9.27%、28.22%),R5020組SKOV3和HO8910細(xì)胞系增殖抑制率分別為(40.36%、53.15%),R5020+U0126組SKOV3和HO8910細(xì)胞系增殖抑制率分別為(29.65%、38.66%)。R5020組兩株細(xì)胞的增殖抑制率均高于R5020+U0126組,R5020+U0126組兩株細(xì)胞的增殖抑制率均高于U0126組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。(3)對(duì)照組SKOV3和HO8910細(xì)胞系凋亡率分別為(9.79%、9.57%),U0126組SKOV3和HO8910細(xì)胞系凋亡率分別為(12.37%、12.62%),R5020組SKOV3和HO8910細(xì)胞系凋亡率分別為(21.66%、27.82%),R5020+U0126組SKOV3和HO8910細(xì)胞系凋亡分別為(15.35%、14.11%)。R5020組兩株細(xì)胞凋亡率均高于對(duì)照組和R5020+U0126組,R5020+U0126組凋亡率均高于U0126組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:孕激素可使卵巢癌HO8910和SKOV3細(xì)胞中PR Ser294磷酸化相對(duì)表達(dá)增加。卵巢癌SKOV3和HO8910細(xì)胞系PR Ser294磷酸化增加后,孕激素抑制卵巢癌細(xì)胞系增殖和促凋亡作用增強(qiáng)。
[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 鹵0.002, respectively. 0.007 鹵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 鹵0.001t0. 006 鹵0. 001C, respectively. No phosphorylation occurred in PRA Ser294. The relative phosphorylation of PRB Ser294 in R5020 SKOV3 and HO8910 cells was 0.72 鹵0.028 and 0.68 鹵0.045, respectively. The relative phosphorylation level of PRA Ser294 was 0.18 鹵0.027 + 0.19 鹵0.028). The relative phosphorylation of PRB Ser294 in R5020 U0126 SKOV3 and HO8910 cell lines was 0.52 鹵0.033, respectively. 0.37 鹵0.012; The relative phosphorylation level of PRA Ser294 in HO8910 cell line was 0.14 鹵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.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.31

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