自噬在声动力疗法抑制C6胶质瘤细胞增殖中的作用  

The Role of Autophagy in the Inhibition of Proliferation of C6 Glioma Cells by Sonodynamic Therapy

作　　者：王来藏[1] 李晨光 谢晨[1] 邵正凯[1] 李建华[1] 李俞辰 王雪峰[1] 张伟光[1] WANG Lai-zang;LI Chen-guang;XIE Chen;SHAO Zheng-kai;LI Jian-hua;LI Yu-chen;WANG Xue-feng;ZHANG Wei-guang(Department of Neurosurgery,The Fourth Affiliated Hospital of Harbin Medical University,Harbin,Heilongjiang,150001,China;Department of Neurosurgery,The Second Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou,Zhejiang,310009,China)

机构地区：[1]哈尔滨医科大学附属第四医院神经外科,黑龙江哈尔滨150001 [2]浙江大学医学院附属第二医院神经外科,浙江杭州310009

出　　处：《现代生物医学进展》2019年第19期3616-3620,3652,共6页Progress in Modern Biomedicine

基　　金：黑龙江省自然科学基金项目(H2016032);国家自然科学基金青年项目(81702462)

摘　　要：目的:探讨自噬在血卟啉单甲醚(Hematoporphyrin monomethyl ether,HMME)介导的声动力疗法(Sonodynamic therapy,SDT)抑制C6胶质瘤细胞增殖中的作用。方法:选取对数期生长的C6胶质瘤细胞并随机分为四组:对照组(未予处理)、超声组(单独超声照射)、HMME组(单独加入HMME)、SDT组(超声照射+HMME)。透射电镜观察SDT处理的C6胶质瘤细胞中自噬体数量的改变。应用qRT-PCR和免疫印迹分析SDT处理对C6胶质瘤细胞中的LC3、Beclin1、Bcl-2 m RNA及蛋白表达水平的影响。MTT检测C6胶质瘤细胞的活力变化。结果:透射电子显微镜显示SDT组自噬体数量较对照组明显增多。SDT组C6胶质瘤细胞中微管相关蛋白1轻链3 (Microtubule associated protein 1 light chain 3, LC3)、Beclin1 m RNA和蛋白水平高于对照组,B细胞淋巴瘤-2(B cell lymphoma-2, Bcl-2) m RNA和蛋白水平低于对照组。与对照组相比,SDT组C6胶质瘤细胞存活率从0 h至6 h逐渐下降,从12 h至72 h逐渐升高。3-甲基腺嘌呤(3-Methyladenine,3-MA)+SDT、氯喹(Chloroquine,CQ)+SDT处理后C6胶质瘤细胞存活率较SDT组明显降低。结论:SDT可能通过诱导自噬抑制C6胶质瘤细胞增殖。Objective: To investigate the role of autophagy in Hematoporphyrin monomethyl ether(HMME)-mediated Sonodynamic therapy(SDT) in inhibiting the proliferation of C6 glioma cells. Methods: C6 glioma cells grown in log phase were randomly divided into four groups: control group(not treated), ultrasound group(individual ultrasound irradiation), HMME group(HMME alone),SDT group(ultrasound irradiation + HMME). The number of autophagic vacuoles in SDT treated C6 glioma cells was observed by transmission electron microscopy. The effects of SDT treatment on the expression of LC3, Beclin1, Bcl-2 in C6 glioma cells were analyzed by q RT-PCR and Western blot. MTT was used to detect the viability of C6 glioma cells. Results: Transmission electron microscopy analysis demonstrated that the number of autophagy in SDT group was significantly higher than that in the control group. The m RNA and protein levels of microtubule associated protein 1 light chain 3(LC3) and Beclin1 in C6 glioma cells of SDT group were higher than those of the control group, and the levels of B cell lymphoma-2(Bcl-2) m RNA and protein were lower than those of the control group. Compared with the control group, the viability of C6 glioma cells in SDT group decreased from 0 h to 6 h, and increased from 12 h to 72 h. The viability of C6 glioma cells after 3-methyladenine(3-MA)+SDT and Chloroquine(CQ)+SDT treatment was significantly lower than that of SDT group. Conclusion: SDT may inhibit the proliferation of C6 glioma cells by inducing autophagy.

关 键 词：自噬 声动力疗法 C6胶质瘤细胞 微管相关蛋白1轻链3 BECLIN1 

分 类 号：R-33[医药卫生] R739.4