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目的探讨氯喹(CQ)对小鼠移植性肉瘤S_(180)的抑瘤作用及其可能机制。方法采用40只S_(180)荷瘤小鼠分为模型组,氯喹低剂量组,氯喹中剂量组,氯喹高剂量组进行体内抑瘤实验,观察不同浓度氯喹对小鼠肉瘤S_(180)的抑制作用;透射电子显微镜观察氯喹作用下荷瘤鼠肿瘤细胞超微结构的变化;流式细胞术检测氯喹诱导荷瘤鼠S_(180)细胞凋亡的情况;免疫组织化学方法检测相关凋亡因子Bcl-2、细胞色素C和Cle-Caspase-3的表达情况;Western blotting检测荷瘤组织Bcl-2和Cle-Caspase-3蛋白表达水平的变化以及线粒体和细胞质中细胞色素C蛋白含量的变化。结果与模型组相比较,氯喹处理组小鼠移植肉瘤S_(180)生长速度显著减慢,肿瘤体积和瘤质量明显减小(P<0.05);透射电子显微镜观察发现,与模型组比较氯喹处理组肿瘤细胞形态出现明显凋亡损伤改变,凋亡小体形成;各剂量组氯喹均可诱导荷瘤鼠S_(180)细胞凋亡,使抗凋亡因子Bcl-2表达下调,凋亡因子Cle-Caspase-3表达上调(P<0.05);并且氯喹能够降低线粒体内细胞色素C蛋白的表达,提高细胞质细胞色素C蛋白的表达(P<0.05),促使线粒体内细胞色素C向细胞质内释放。结论氯喹能够抑制小鼠S_(180)肉瘤的生长,可能是通过线粒体凋亡途径诱导细胞凋亡,从而发挥抑瘤的作用。
Objective To investigate the anti-tumor effect of chloroquine (CQ) on the transplanted sarcoma S_ (180) in mice and its possible mechanism. Methods Forty S 180 tumor-bearing mice were divided into model group, low-dose chloroquine group, middle-dose chloroquine group and high-dose chloroquine group for antitumor activity in vivo. The effects of different concentrations of chloroquine on S 180 The changes of ultrastructure of tumor-bearing mice tumor cells under the action of chloroquine were observed by transmission electron microscopy. The apoptosis of S 180 cells induced by chloroquine was detected by flow cytometry. The apoptosis of tumor cells was detected by immunohistochemistry Bcl-2, Cytochrome C and Cle-Caspase-3 were detected by Western blotting. The changes of Bcl-2 and Cle-Caspase-3 protein expression and the changes of cytochrome C protein in mitochondria and cytoplasm were detected by Western blotting. Results Compared with the model group, the growth rate of S_ (180) in the mice treated with chloroquine significantly slowed down and the tumor volume and tumor mass decreased significantly (P <0.05). Transmission electron microscopy revealed that compared with the model group, The morphological changes of apoptotic cells were observed and the apoptotic bodies were formed. Chloroquine of each dose group could induce the apoptosis of S_ (180) cells and down-regulate the expression of anti-apoptotic factor Bcl-2 (P <0.05). And chloroquine could reduce the expression of cytochrome C protein in mitochondria and increase the expression of cytochrome C protein in cytoplasm (P <0.05), and promote the release of cytochrome C in the mitochondria. Conclusion Chloroquine can inhibit the growth of S180 sarcoma in mice, which may induce cell apoptosis through mitochondrial apoptosis pathway and thus exert antitumor effect.