腺苷酸活化蛋白激酶激活与卵巢癌细胞顺铂耐药的关系

Relationship between AMPK activation and cisplatin resistance in the ovarian cancer cells

目的:研究腺苷酸活化蛋白激酶(AMPK)激活与卵巢癌细胞对顺铂耐药的关系。方法:以不同浓度的顺铂(0,3.125,6.25,12.5,25,50μmol·L-1)分别作用人卵巢癌细胞SKOV3及其顺铂耐药株SKOV3/DDP 24h,MTT法检测顺铂的抑制率;透射电镜检测SKOV3和SKOV3/DDP中的自噬体;流式细胞术检测顺铂作用SKOV3和SKOV3/DDP的凋亡率;Western blot法测定基础状态下及顺铂单独或联合AMPK抑制剂compound C作用时,SKOV3和SKOV3/DDP中p-AMPK、自噬微管相关蛋白轻链3(LC3-Ⅱ)和凋亡蛋白聚腺苷二磷酸-核糖聚合酶(PARP)的表达差异;并测定compound C联合顺铂作用SKOV3和SKOV3/DDP时的细胞活性。结果:与SKOV3比较,SKOV3/DDP对顺铂耐药(RI=6.8);SKOV3/DDP中自噬体明显增多(P<0.01);顺铂对SKOV3/DDP早期凋亡率(7.0±0.55)%远低于SKOV3早期凋亡率(30.5±0.34)%(P<0.01);SKOV3/DDP中p-AMPK、LC3-Ⅱ表达量均增加,顺铂作用时Cleaved-PARP表达不明显;compound C与顺铂同时作用时,SKOV3/DDP中p-AMPK、LC3-Ⅱ表达减少,Cleaved-PARP表达水平显著升高,且SKOV3/DDP对顺铂敏感性增加。结论:SKOV3/DDP中p-AMPK的激活诱导卵巢癌细胞自噬可能导致其对顺铂耐药,compound C抑制p-AMPK的激活,与顺铂同时作用促进顺铂诱导SKOV3/DDP细胞凋亡,且增加其对顺铂的敏感性。

ONECTIVE To study the relationship between AMPK activation and cisplatin resistance in tile ovarian cancer cells. METHODS SKOV3 and SKOV3/DDP cells were stimulated by cisplatin at different concentrations （（I,3. 125,6. 25, 12.5,25 and 50/lmol.L-1 ） for 24 h. The inhibition rate of cisplatin was determined by MTT assay. Formalion of aulophagy was detected by transmission electron microscopy. Apoptosis rate induced by cisplatin was analysized by flow cytometry. Pro rein levels of LC3- 11 , PARP and p-AMPK were determined by Western blot. RIESUL]N Compared with SKi）V3, SKOV3/DDP was significantly resistant to cisplatin （R] = 6. 8）. Apoptosis rate was （7. 0 ±0.55） %, lower than that of SKOV3 （30.5 ± 0. 34）% （P〈0. 01 ）. Protein levels of p-AMPK and I.C3- II were increased in SK（）V3/DDP, but reduced by compound （7. Inhi bition of p-AMPK by compound C enhanced apoptosis rate and inhibition rate in SKOV3/DDP induced by cisplalin. CONCI.U- SION Resistance of ovarian cancer cells may be associated with activation of AMPK in SKOV3/DDP. AMPK inhibitor corn pound C can inhibit activation of AMPK and decrease expression of LC3-Ⅱ , but promote apoptosis induced by cisplatin in owLri an cancer cells,and increase the eisplatin sensitivity to cisplatin resistant strains.