本文選題：哌立福辛 + 順鉑��； 參考：《現(xiàn)代婦產(chǎn)科進(jìn)展》2015年10期

【摘要】：目的:探討哌立福辛(perifosine)與順鉑(cisplatin)聯(lián)合對(duì)人卵巢透明細(xì)胞癌ES2細(xì)胞系凋亡及Caspase-3和PARP表達(dá)的影響。方法:以不同濃度(0、2、4、6、8和10μmol/L)哌立福辛作用于人卵巢透明細(xì)胞癌ES2細(xì)胞系48h。CCK-8法檢測(cè)細(xì)胞增殖抑制情況。以哌立福辛最小有作用劑量與不同濃度(0、10、40、70和100μmol/L)順鉑聯(lián)合處理細(xì)胞48h,以金氏公式篩選出協(xié)同抑制作用最明顯的聯(lián)合劑量,以該聯(lián)合劑量處理ES2細(xì)胞48h后,DAPI染色法、流式細(xì)胞儀檢測(cè)細(xì)胞凋亡情況;用Giemsa染色檢測(cè)細(xì)胞有絲分裂指數(shù);Western blot法檢測(cè)細(xì)胞中Caspase-3(cleaved)及PARP(cleaved)蛋白的表達(dá)情況。結(jié)果:CCK-8法檢測(cè)提示,哌立福辛作用48h時(shí)對(duì)ES2細(xì)胞的最小作用劑量為4μmol/L。金氏公式計(jì)算結(jié)果提示,4μmol/L哌立福辛+40μmol/L順鉑聯(lián)合用藥協(xié)同抑制細(xì)胞增殖作用最強(qiáng)(q=1.21);與哌立福辛或順鉑單獨(dú)作用相比,DAPI染色發(fā)現(xiàn)聯(lián)合用藥組鏡下可見(jiàn)大量的核碎裂及凋亡小體;流式細(xì)胞儀檢測(cè)發(fā)現(xiàn),聯(lián)合用藥能協(xié)同增加細(xì)胞凋亡率(q=1.32,P0.01)。Giemsa染色結(jié)果顯示,聯(lián)合用藥能協(xié)同抑制細(xì)胞有絲分裂(P0.01)。Western blot檢測(cè)發(fā)現(xiàn),聯(lián)合用藥能明顯增加細(xì)胞中Caspase-3(cleaved)蛋白的表達(dá),降低PARP(cleaved)蛋白的表達(dá)。結(jié)論:哌立福辛與順鉑聯(lián)合能協(xié)同通過(guò)增加Caspase-3(cleaved)蛋白表達(dá)及降低PARP(cleaved)蛋白表達(dá),促進(jìn)人卵巢透明細(xì)胞癌ES2細(xì)胞發(fā)生凋亡。
[Abstract]:Aim: to investigate the effects of pirifampicin (perifosine) combined with cisplatin (cisplatin) on apoptosis and expression of Caspase-3 and PARP in human ovarian clear cell carcinoma cell line ES2. Methods: the proliferation inhibition of human ovarian clear cell carcinoma ES2 cell line was detected by different concentrations of pirifoxine (0 渭 mol / L and 10 渭 mol / L) by CCK-8 method. The cells were treated with pirifoxine at a minimum active dose of 4070 and 100 渭 mol / L cisplatin for 48 h. The most obvious synergistic inhibitory dose was screened by Kim's formula. The cells were treated with the combined dose for 48 h and then treated with DAPI staining method. The cell apoptosis was detected by flow cytometry and the expression of Caspase-3 (cleaved) and PARP (cleaved) protein was detected by Giemsa staining and Western blot assay. Results the minimum dose of pirifoxine on ES2 cells was 4 渭 mol / L after 48h exposure. The results of Kim's formula showed that 4 渭 mol / L pirifoxine 40 渭 mol / L cisplatin combined with cisplatin had the strongest synergistic inhibitory effect on cell proliferation (q1. 21), compared with pirifampicin or cisplatin alone, a large number of nuclear fragmentation and apoptotic corpuscles were found in the combined treatment group. The results of flow cytometry showed that the combined drug could increase the cell apoptosis rate (QN 1.32 P0.01). Giemsa staining results showed that the combined drug could inhibit the cell mitosis (P0.01). Western blot assay showed that the combined treatment could significantly increase the expression of Caspase-3 (cleaved) protein in the cells. The expression of PARP (cleaved) protein was decreased. Conclusion: the combination of pirifampicin and cisplatin can promote apoptosis of human ovarian clear cell carcinoma ES2 cells by increasing the expression of Caspase-3 (cleaved) protein and decreasing the expression of (cleaved) protein.
【作者單位】： 中國(guó)醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院北京協(xié)和醫(yī)院婦產(chǎn)科;
【分類號(hào)】：R737.31

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本文編號(hào)：2059717