力达霉素诱导人宫颈癌Caski 细胞发生免疫原性细胞凋亡

目的:探讨抗肿瘤药物力达霉素能否抑制人宫颈癌Caski 细胞增殖并且诱导Caski 细胞发生免疫原性细胞凋亡。方法:MTT 比色法检测Caski 细胞增殖;Annexin V-FITC/ PI 双染流式细胞术检测Caski 细胞凋亡。Western blot 分析细胞中Bax 和Bcl-2 的表达;免疫荧光流式细胞术检测细胞凋亡后细胞膜上钙网蛋白的表达。结果:力达霉素抑制Caski 细胞的增殖,具有时间和剂量依赖性;5 g/ L 的力达霉素作用Caski 细胞48 h 后,肿瘤细胞凋亡率为11.5%;Bax 表达量显著增加,而Bcl-2 含量明显减少(P<0.05);细胞膜表面钙网蛋白表达高达67.2%,显著高于对照组的2.31%。结论:力达霉素能够有效抑制人宫颈癌Caski 细胞增殖,其作用机制与线粒体途径诱导细胞凋亡相关,同时促使钙网蛋白转移到Caski 细胞膜表面表达,可能具有诱导肿瘤细胞发生免疫原性细胞凋亡的能力。

Immunogenic apoptosis of human Caski cervical cancer cells induced by Lidamycin

Objective:To investigate effects of Lidamycin (LDM,C-1027) on the proliferation and immunogenic transform of human Caski cervical cancer cells and to provide the basic experiment data and theoretical supports for establishment of the new immunotherapy method mediated by LDM.Methods: MTT was used for the analysis of cell proliferation;apoptosis rate was analyzed by flow cytometry;Western blot was used to analyze the effect of LDM on the expression of Bax and Bcl-2 in Caski cells;the Flow cytometry was used to detect the expression of Calreticulin (CRT) on the cell surface.Results: Lidamycin inhibited proliferation of Caski cells significantly in the time-and dose-dependent manners;The apoptotic cell ratio induced by 5 g/ L Lidamycin was 11.5% ,Comparing with the control group, Lidamycin treatment increased Bax but decreased Bcl-2 contents significantly within Caski cells, it also significantly increased the expression of CRT on the cell surface of Caski cells from 2.31% to 67.2% .Conclusion: Lidamycin has pharmacological activity in inhibiting proliferation of the human cervical Caski cells and the underlying mechanism is related with inducing the intrinsic mitochondrial pathway of apoptosis.In the same time,Lidamycin can increase the expression of CRT on the cell surface,so it may have the ability to promote the immunogenic apoptosis of tumor cells.