CAJ | 학술논문

目的：探讨内质网应激（ ERS）在卵巢癌顺铂耐药中的作用及其机制。方法沙奎那韦诱导肿瘤细胞SKOV3后，采用逆转录聚合酶链反应（ RT-PCR）法和Western blot 法检测SKOV3细胞中哺乳动物雷帕霉素靶蛋白（ mTOR）、Beclin1 mRNA及其蛋白的表达，四甲基偶氮唑蓝（ MTT）法检测沙奎那韦对卵巢癌细胞顺铂敏感性的影响。结果顺铂对SKOV3细胞的半数抑制浓度（ IC50）为（5．490±1．148）μg/ml。当10、20μmol/L沙奎那韦作用SKOV3细胞后，顺铂的IC50分别为（11．199±0．984）μg/ml和（14．906±2．015）μg/ml，提示卵巢癌细胞对顺铂敏感性下降，差异有统计学意义（P＝0．001）。对照组、顺铂组、沙奎那韦＋顺铂组、LY294002组和沙奎那韦组中 SKOV3细胞的mTOR、Beclin1 mRNA及其蛋白表达水平不同，差异有统计学意义（均P＜0．001）。 mTOR、Beclin1 mRNA相对表达水平在沙奎那韦＋顺铂组中最高，分别为0．684±0．072和0．647±0．047；其次为沙奎那韦组，分别为0．577±0．016和0．565±0．037。 mTOR、Beclin1蛋白相对表达水平在沙奎那韦＋顺铂组中最高，分别为0．624±0．058和0．924±0．033；其次为沙奎那韦组，分别为0．544±0．019和0．712±0．024。以3-甲基腺嘌呤抑制肿瘤细胞自噬，可阻断沙奎那韦诱导SKOV3细胞顺铂敏感性下降（ P＜0．001）。结论沙奎那韦可有效诱导SKOV3细胞ERS，ERS可导致卵巢癌细胞顺铂敏感性下降，其机制可能与ERS通过调节mTOR和Beclin1表达而改变肿瘤细胞自噬水平有关。肿瘤细胞ERS与细胞自噬可能成为提高肿瘤化疗疗效及逆转耐药的新靶点。
목적：탐토내질망응격（ ERS）재란소암순박내약중적작용급기궤제。방법사규나위유도종류세포SKOV3후，채용역전록취합매련반응（ RT-PCR）법화Western blot 법검측SKOV3세포중포유동물뢰파매소파단백（ mTOR）、Beclin1 mRNA급기단백적표체，사갑기우담서람（ MTT）법검측사규나위대란소암세포순박민감성적영향。결과순박대SKOV3세포적반수억제농도（ IC50）위（5．490±1．148）μg/ml。당10、20μmol/L사규나위작용SKOV3세포후，순박적IC50분별위（11．199±0．984）μg/ml화（14．906±2．015）μg/ml，제시란소암세포대순박민감성하강，차이유통계학의의（P＝0．001）。대조조、순박조、사규나위＋순박조、LY294002조화사규나위조중 SKOV3세포적mTOR、Beclin1 mRNA급기단백표체수평불동，차이유통계학의의（균P＜0．001）。 mTOR、Beclin1 mRNA상대표체수평재사규나위＋순박조중최고，분별위0．684±0．072화0．647±0．047；기차위사규나위조，분별위0．577±0．016화0．565±0．037。 mTOR、Beclin1단백상대표체수평재사규나위＋순박조중최고，분별위0．624±0．058화0．924±0．033；기차위사규나위조，분별위0．544±0．019화0．712±0．024。이3-갑기선표령억제종류세포자서，가조단사규나위유도SKOV3세포순박민감성하강（ P＜0．001）。결론사규나위가유효유도SKOV3세포ERS，ERS가도치란소암세포순박민감성하강，기궤제가능여ERS통과조절mTOR화Beclin1표체이개변종류세포자서수평유관。종류세포ERS여세포자서가능성위제고종류화료료효급역전내약적신파점。
Objective The study intended to investigate the effect and mechanism of endoplasmic reticulum stress on cisplatin resistance in ovarian carcinoma .Methods RT-PCR and Western blot were used to test the expression of mTOR and Beclin 1 mRNA and protein in ovarian cancer SKOV 3 cells after saquinavir induction .MTT assay was used to analyze the influence of saquinavir on cisplatin sensitivity in SKOV3 cells.Results The IC50 of SKOV3 cells was ( 5.490 ±1.148 ) μg/ml.After induced by Saquinavair 10μmol/L and 20μmol/L, the IC50 of SKOV3 cells was increased to (11.199 ±0.984) μg/ml and (14.906 ±2.015) μg/ml, respectively.It suggested that the sensitivity of ovarian cancer cells to cisplatin was decreased significantly (P=0.001).The expression of mTOR and Beclin1 mRNA and protein was significantly different among the five groups: the (Saquinavair +DDP) group of, Saquinavair group, LY294002 group, DDP group and control group(P<0.001).The expressions of mTOR and Beclin1 mRNA were highest in the ( Saquinavair +DDP ) group, 0.684 ±0.072 and 0.647 ±0.047, respectively;Secondly, the Saquinavair group , 0.577 ±0.016 and 0.565 ±0.037, respectively.The expressions of mTOR and Beclin1 proteins were also highest in the (Saquinavair+DDP) group, 0.624 ±0.058 and 0.924 ± 0.033, respectively, followed by the Saquinavair group , 0.544 ±0.019 and 0.712 ±0.024.3-MA inhibited the autophagy and restored cisplatin sensitivity in the SKOV 3 cells after Saquinavir induced ER stress ( P<0.001 ).Conclusions Saquinavir can effectively induce endoplasmic reticulum stress in SKOV 3 cells. Endoplasmic reticulum stress can decrease the sensitivity to cisplatin in SKOV 3 cells.The mechanism of the decrease of sensitivity to cisplatin in SKOV 3 cells may be that ERS regulates cell autophagy through the mTOR and Beclin1 pathways .ERS of tumor cells and autophagy may become a new target to improve the therapeutic effect of chemotherapy and to reverse the drug resistance in tumor treatment .