摘要
目的:通过上调Sonic Hedgehog(Shh)信号通路关键因子Gli1,探究Shh信号通路与食管癌细胞放射抗拒性及细胞周期的关系。方法:用过表达Gli1质粒转染人食管癌Eca109细胞,记为Eca109-ox-Gli1组;转染空载质粒为阴性对照组;以不作处理的亲本细胞株为正常对照组。Real-time PCR与Western blot检测转染后细胞内Gli1的表达,集落形成实验检测细胞放射敏感性,流式细胞术检测转染前后细胞周期分布。结果:Real-time PCR与Western blot结果均显示Eca109-ox-Gli1组的Gli1表达显著高于阴性对照组和正常对照组(P<0.05)。集落形成实验显示2 Gy射线照射后,Eca109-ox-Gli1组的存活分数较正常对照组明显升高(P<0.05),转染后细胞对射线敏感性降低,放射抗拒性增加。Eca109-ox-Gli1组中S期细胞分布比例显著高于对照组(P<0.01),再照射6Gy后,3组细胞均出现了G_2/M期阻滞,正常对照组相比于Eca109-ox-Gli1组G_2/M期细胞分布显著增高(P<0.01)。结论 :上调Shh信号通路关键因子Gli1能够增强食管癌细胞的放射抗拒性,这一过程可能是通过调控细胞周期实现的。
AIM： To investigate the relationship between Sonic Hedgehog（ Shh） signaling pathway and cell cycle and radioresistance of esophageal cancer by up-regulating Gli1,a key factor in Shh signaling pathway. METHODS：The human esophageal cancer cell line Eca109 was transfected with plasmid to induce Gli1 over-expression,which served as Eca109-ox-Gli1 group. In addition,Eca109 cells transfected with empty plasmid served as negative control group and the untreated Eca109 cells were used as normal control group. The expression of Gli1 was confirmed by real-time PCR and Western blot. The radiosensitivity of the cells in the 3 groups was determined by colony formation assay. The effect of irradiation on the cell cycle was analyzed by flow cytometry. RESULTS： The expression of Gli1 in Eca109-ox-Gli1 group was higher than that in the other 2 groups（ P〈0. 05）. The survival fraction at dose of 2 Gy in Eca109-ox-Gli1 group was higher than that in normal control group,indicating that the radioresistance of the Eca109 cells transfected with Gli1 plasmid was increased. The cells in Eca109-ox-Gli1 group showed higher S phase proportion than that in normal control group and negative control group（ P〈0. 01）. After irradiation at dose of 6 Gy,all cells in the 3 groups found that the cell cycle was arrested at G2/M phase,while the cells in normal control group showed higher G2/M phase proportion than that in Eca109-ox-Gli1 group（ P〈0. 01）. CONCLUSION： The up-regulation of Gli1 may enhance the radioresistance of esophageal cancer by regulating the cell cycle.
作者
余今菁
贺昱霖
刘佳琪
姚菲
刘群
龙辉
吴清明
YU Jin-jing;HE Yu-lin;LIU Jia-qi;YAO Fei;LIU Qun;LONG Hui;WU Qing-ming(School of Medicine;Hubei Provincial Key Laboratory of Occupational Hazard Identification and Control;Department of Gastroenterology of Affiliated Tianyou Hospital, Wuhan University of Science and Technology, Wuhan 430065, China;Institute of Biomedical Research, Taihe Hospital, Hubei University of Medicine, Shiyan 442000, China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2018年第5期799-803,共5页
Chinese Journal of Pathophysiology
基金
湖北省自然科学基金资助项目(No.2014 CFB818)
湖北省领军人才培养工程
