棉酚联合放疗诱导宫颈癌HeLa细胞凋亡的研究

Study on the effect of gossypol combined with radiotherapy in inducing apoptosis of human cervical carcinoma HeLa cells

目的:研究棉酚(gossypol)联合放疗对宫颈癌HeLa细胞的增殖抑制与促细胞凋亡作用,以探讨棉酚的抗肿瘤作用机制。方法:利用MTT法检测棉酚对HeLa细胞生长抑制率的影响;采用丫啶橙/溴化乙锭双染色法、FCM观察细胞凋亡形态并检测细胞凋亡率;免疫细胞化学法检测Bcl-2和Bcl-xL蛋白的表达水平。结果:棉酚对HeLa细胞增殖有明显的抑制作用,且呈剂量及时间依赖性;棉酚、放疗、棉酚联合放疗能明显诱导HeLa细胞凋亡,且棉酚联合放疗组的诱导作用明显优于2者单独处理组。棉酚、放疗和棉酚联合放疗均可降低HeLa细胞内Bcl-2和Bcl-xL蛋白的表达水平,但棉酚联合放疗组的作用弱于2者单独处理组。结论:棉酚可抑制宫颈癌HeLa细胞的增殖并诱导细胞凋亡,可能增加放疗对宫颈癌HeLa细胞的敏感性。

Objective：To investigate the effect of gossypol and gossypol-based combination radiotherapy on the proliferation and apoptosis of cervical carcinoma HeLa cells and explore its anti-tumor mechanism.Methods：The inhibitory effect of gossypol on prolife-ration of HeLa cells was measured by MTT assay.The morphology of apoptotic cells was observed by the acridine orange/ethidium bromide（AO/EB） double staining and the apoptotic ratio was determined by flow cytometry.The expressions of Bcl-2 and Bcl-xL proteins were observed by immunocytochemistry.Results：Gossypol significantly inhibited the proliferation of HeLa cells in a dose-and time-dependent manner.Gossypol and radiotherapy alone or in combination induced apoptosis of HeLa cells significantly.The effect of combination therapy was superior to that treated with gossypol or radiotherapy alone.Gossypol treatment and radiotherapy alone or in combination down-regulated expression of Bcl-2 and Bcl-xL proteins.The effect of combined therapy in down-regulating Bcl-2 and Bcl-xL was stronger than that induced by gossypol or radiotherapy alone.Conclusion：Gossypol effectively inhibits the proliferation and induces apoptosis of cervical carcinoma HeLa cells and enhances the sensitivity of HeLa cells to radiotherapy.