TWEAK调控凋亡增加上皮性卵巢癌细胞对铂类药物敏感性的研究

TWEAK enhances cisplasin sensitivity in human epithelial ovarian cancer cells by regulating apoptosis.

目的:探讨肿瘤坏死因子样凋亡微弱诱导剂(TWEAK)在调控上皮性卵巢癌(EOC)细胞对顺铂(DDP)敏感性中发挥的作用。方法:不同浓度TWEAK刺激A2780/cDDP细胞后,采用CCK-8法检测细胞的增殖能力及DDP半数抑制浓度(IC50)的变化;采用流式细胞术检测细胞凋亡率的变化;采用透射电镜检测细胞核染色质形态和凋亡小体的变化;Western blot法检测细胞凋亡相关蛋白caspase 3的活化情况。结果:TWEAK对A2780/cDDP细胞的增殖能力无影响,但能显著增加A2780/cDDP细胞对DDP的敏感性。流式结果显示,随着TWEAK剂量增加,A2780/cDDP细胞的凋亡显著增加。TWEAK刺激后A2780/cDDP细胞中caspase 3活化体表达增加,细胞核染色质发生形态变化,诱导了细胞的凋亡及凋亡小体形成。结论:TWEAK通过活化凋亡相关通路,促进EOC细胞凋亡,提高了DDP耐药EOC细胞对DDP的敏感性。这可能为临床开创逆转EOC铂类耐药的治疗新途径提供理论依据。

Objective：To investigate the effects of TWEAK on reguluating cisplatin sensitivity in human ovarian cancer cell. Methods：The half cell lethal dose（IC50） of cisplatin in A2780/cDDP cells after stimulation by 100ng/ml of TWEAK for 24 hours was assayed by CCK-8 kit. The changes of apoptosis ratio before or after TWEAK treatment were determined by flow cytometry. The protein expression of caspase3 after different concentration of TWEAK treat-ment was detected by Western blot assay. The representative images of nuclear chromatin mor-phology and apoptotic body were investigated by transmission electron microscopy. Results： TWEAK had no effect on the proliferation of A2780/cDDP cells. However, A2780/cDDP cells were more sensitive to cisplatin after TWEAK treatment. The apoptosis ratio and the protein ex-pression of the cleaved caspase 3 in TWEAK treated-A2780/cDDP cell were significantly in-creased in a dose-dependent manner. Meanwhile, the images of nuclear chromatin morphology changed and apoptotic body were detected in A2780/cDDP cell treated with 100ng/ml of TWEAK for 24h by transmission electron microscopy. Conclusions ：TWEAK may increase che-mosensitivity to cisplatin by activating apoptosis pathway in chemoresistant human ovarian canc-er cell lines A2780/cDDP. Our data may provide theoretical basis for developing new therapy for cisplatin resistant ovarian cancer.