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自噬诱导是肿瘤细胞对化疗药物抵抗性的原因之一,该研究探讨溶酶体抑制剂氯喹对喜树碱(camptothecin,CPT)诱导的宫颈癌细胞Si Ha死亡的增敏效果。CPT和/或氯喹处理宫颈癌Si Ha细胞,MTT法检测细胞增殖,DAPI和TUNEL染色观察细胞凋亡,Western blot和免疫荧光检测自噬及凋亡相关蛋白。结果发现,CPT处理后,Si Ha细胞MAP1LC3B荧光点和LC3II(microtubuleassociated protein light chain 3II)蛋白水平增加,p62荧光点和蛋白质水平则减少;而采用氯喹特异抑制自噬后,可明显提高CPT诱导的细胞凋亡、caspase-9的激活和PARP(poly ADP-ribose polymerase)的切割,而全长caspase-2水平显著下降。以上结果提示,氯喹可通过抑制细胞自噬而增强宫颈癌细胞株Si Ha对CPT诱导细胞凋亡的敏感性。
Induction of autophagy is one of the reasons why tumor cells respond to chemotherapeutic drugs. The aim of this study was to investigate the sensitization effect of lysosome inhibitor chloroquine to the death of Si Ha cells induced by camptothecin (CPT) in cervical cancer cells. Cervical carcinoma Si Ha cells were treated with CPT and / or chloroquine, cell proliferation was detected by MTT assay, apoptosis was observed by DAPI and TUNEL staining, and autophagy and apoptosis related proteins were detected by Western blot and immunofluorescence. The results showed that, after CPT treatment, SiH cells MAP1LC3B fluorescence point and LC3II (microtubuleassociated protein light chain 3II) protein levels increased, p62 fluorescence point and protein levels decreased; while using chloroquine specific inhibition of autophagy, can significantly increase the CPT-induced Apoptosis, activation of caspase-9 and cleavage of PARP (poly ADP-ribose polymerase), while the full-length caspase-2 level was significantly decreased. The above results suggest that chloroquine can enhance the sensitivity of cervical cancer cell line Si Ha to CPT-induced apoptosis by inhibiting autophagy.