洛铂对人胆管癌RBE细胞凋亡显微结构及超微结构的作用观察

Effect of Lobaplatin on Apoptosis of Human Cholangiocarcinoma Cells Line RBE:Histochemical and Electron Microscopic Observations

目的观察洛铂诱导人胆管癌RBE细胞凋亡显微结构及超微结构的变化,探讨洛铂对诱导胆管癌细胞凋亡作用.方法体外培养RBE-TCHU179人胆管癌细胞,给予不同剂量洛铂处理,瑞氏-姬姆萨染色及AO/EB染色观察细胞形态学和计数细胞凋亡百分率,扫描电镜和透射电镜观察细胞表面超微结构和细胞内超微结构的变化.结果 AO/EB荧光染色观察到洛铂作用24 h后,随洛铂剂量加大,人胆管癌RBE细胞凋亡百分率升高,25μg/mL作用凋亡细胞占47.8%.光镜和扫描电镜可见细胞形态学呈现典型的凋亡变化,可见凋亡小体的形成.透射电镜可观察到细胞核呈新月状改变.结论洛铂能引起人胆管癌RBE细胞凋亡显微结构及超微结构改变,可能与细胞凋亡和侵袭力有关.

Objective The purpose of this study was to observe the effect of lobaplatin on human cholangiocarcinoma cells line RBE by histochemical and electron microscopic, for investigating the mechanism of lobaplatin-induced RBE cells apoptosis. Methods The RBE cells were cultured in vitro,followed by treating with various concentration of lobaplatin for 24 h. Wright-Giemsa staining and AO/EB staining were used to determine the inhibition rate and microstructure changes. And the ultrastructural changes were studied by scanning electron microscopy and transmission electron microscopy. Res ults AO/EB staining showed that the apoptosis rate of RBE cells increased with increasing dose of lobaplatin. After treatment by 25 μg/mL lobaplatin for 24 h, the apoptotic cells accounted for 47.8%. Light microscopy and scanning electron microscopy showed typical apoptotic changes, and the formation of apoptotic bodies was observed. The nucleus crescent-like change also could be observed by TEM. Conclus ion Lopaplatin can induce the morphological and ultrastructural changes on RBE cell line,which may be closely related to apoptosis and invasion. The results provide a valuable reference for the further study of lobaplatin antitumor effect.