【摘要】：目的:研究腺苷酸活化蛋白激酶(AMPK)激活與卵巢癌細胞對順鉑耐藥的關系。方法:以不同濃度的順鉑(0,3.125,6.25,12.5,25,50μmol·L-1)分別作用人卵巢癌細胞SKOV3及其順鉑耐藥株SKOV3/DDP 24h,MTT法檢測順鉑的抑制率;透射電鏡檢測SKOV3和SKOV3/DDP中的自噬體;流式細胞術檢測順鉑作用SKOV3和SKOV3/DDP的凋亡率;Western blot法測定基礎狀態(tài)下及順鉑單獨或聯(lián)合AMPK抑制劑compound C作用時,SKOV3和SKOV3/DDP中p-AMPK、自噬微管相關蛋白輕鏈3(LC3-Ⅱ)和凋亡蛋白聚腺苷二磷酸-核糖聚合酶(PARP)的表達差異;并測定compound C聯(lián)合順鉑作用SKOV3和SKOV3/DDP時的細胞活性。結(jié)果:與SKOV3比較,SKOV3/DDP對順鉑耐藥(RI=6.8);SKOV3/DDP中自噬體明顯增多(P0.01);順鉑對SKOV3/DDP早期凋亡率(7.0±0.55)%遠低于SKOV3早期凋亡率(30.5±0.34)%(P0.01);SKOV3/DDP中p-AMPK、LC3-Ⅱ表達量均增加,順鉑作用時Cleaved-PARP表達不明顯;compound C與順鉑同時作用時,SKOV3/DDP中p-AMPK、LC3-Ⅱ表達減少,Cleaved-PARP表達水平顯著升高,且SKOV3/DDP對順鉑敏感性增加。結(jié)論:SKOV3/DDP中p-AMPK的激活誘導卵巢癌細胞自噬可能導致其對順鉑耐藥,compound C抑制p-AMPK的激活,與順鉑同時作用促進順鉑誘導SKOV3/DDP細胞凋亡,且增加其對順鉑的敏感性。
[Abstract]:Aim: to study the relationship between adenylate activated protein kinase (AMPK) activation and cisplatin resistance in ovarian cancer cells. Methods: the inhibition rate of cisplatin was detected by different concentrations of cisplatin (03.125 Cisplatin 6.255.12.5U 2550 渭 mol L-1) on human ovarian cancer cell line SKOV3 and cisplatin resistant cell line SKOV3/DDP for 24 h, and the autophagy in SKOV3 and SKOV3/DDP was detected by transmission electron microscope (TEM). Detection of apoptosis rate of SKOV3 and SKOV3/DDP induced by Cisplatin by flow Cytometry and Western blot Assay for p-AMPK, LC3- 鈪，

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