P162抑制Hedgehog信号转录因子Gli-1增强食管癌Eca109细胞的放射敏感性  

P162 enhances radiosensitivity of esophageal carcinoma Eca109 cells by inhibiting Hedgehog signaling transcription factor Gli-1

作　　者：陈洁[1] 吴清明[2] 龙辉[2] 张红[1] 陈建华 

机构地区：[1]武汉科技大学,湖北省武汉市430065 [2]武汉科技大学附属天佑医院,湖北省武汉市430064 [3]武汉凯泰新生物技术有限公司,湖北省武汉市430074

出　　处：《世界华人消化杂志》2014年第5期615-623,共9页World Chinese Journal of Digestology

摘　　要：目的:探讨新药多肽P162通过抑制Hedgehog(H h)信号转录因子G l i-1对人食管癌细胞株Eca109起放射增敏作用.方法:反复多次X线(剂量累计60 Gy)照射Eca109诱导食管癌细胞Eca109R,采用CCK8法测增殖抑制率,免疫细胞化学法、免疫荧光技术测Gli-1的表达,HE染色观察细胞形态学改变,Western blot测核内Gli-1表达及动态监测Eca109放疗后核内Gli-1变化,流式细胞仪测细胞凋亡率.实验分以下4组:未照射加药组、照射加药组、未照射不加药组和照射不加药组,每组中含Eca109、Eca109R两种细胞.结果:Eca109R增殖抑制率明显低于Eca109,具有一定放射抗拒性;Eca109R较Eca109高表达Gli-1(0.45±0.01,0.32±0.01,P<0.0001);Eca109放疗后Gli-1于2 d,14 d表达与对照相比(0.0882±0.011,0.3560±0.015 vs 0.2552±0.0103),差异有统计学意义(P<0.05);20μmol/L P162干预Eca109R、Eca109细胞中,与0μmol/L P162干预比较Gli-1表达下调,分别为:0.2553±0.011,0.2578±0.014(未照射),0.1324±0.012,0.0595±0.011(照射2 d),0.1741±0.013,0.2397±0.112(照射14 d),差异均有统计学意义(P<0.0001),P162联合放疗促细胞凋亡.结论:Hh信号转录因子Gli-1与食管癌放射抗拒相关.P162放射增敏作用可能与抑制转录因子Gli-1、促细胞凋亡相关.AIM： To investigate whether P162 enhances radiosensitivity of esophageal carcinoma cell line Eca109 by inhibiting Hedgehog signaling transcription factor Gli-1. METHODS： Eca109 cells （a total dose of 60 Gy） to induce radioresistant esophageal carcinoma cell line Eca109R. The inhibition of cell proliferation was determined by Cell Counting Kit assay. The expression of Gli-1 was detected by immunohistochemistry and immunofluorescence. HEstaining was employed to observe the changes in cell morphology. Western blot was employed to determine the nuclear expression of Gli-1 and dynamic changes of Gli-1 in irradiated Eca109 cells. Apoptosis was determined by flow cytometry. The following four groups were included in the experiments： untreated cells, P162-treated cells, irradiated cells, and P162-treated irradiated cells. Eca109 and Eca109R cells were included in each group. RESULTS： Eca109R possessing certain radiation resistance displayed lower ability of growth inhibition than Eca109 cells. Nuclear Gli-1 expression was significantly higher in Eca109R cells than in Eca109 cells （0.45 ± 0.01 vs 0.32 ± 0.01, P 〈 0.0001）. On days 2 and 14 after irradiation, the nuclear expression of Gliq in Eca109 cells was higher than that in control cells （0.0882 ± 0.011, 0.3560 ± 0.015 vs 0.2552 ± 0.0103, P 〈 0.05）. In both Eca109R and Eca109 cells, the nuclear expression of Gli-1 was reduced after treatment with 20 μmol/L P162 [0.2553 ± 0.011, 0.2578 ± 0.014 （non-irradiation）; 0.1324 ± 0.012, 0.0595 ± 0.011（2 d after irradiation）; 0.1741 ± 0.013, 0.2397 ± 0.112 （14 d after irradiation）, P 〈 0.0001]. P162 combined with radiotherapy facilitated cells apoptosis. CONCLUSION： Nuclear Gli-1 expression is related to radioresistance of esophageal cancer ceils. P162 enhances radiosensitivity of Eca109 cell possibly by inhibiting Gli-1 expression and promoting apoptosis.

关 键 词：食管癌 HEDGEHOG Gli-1 P162 放射抗拒 

分 类 号：R735.1[医药卫生—肿瘤]