GOLPH3对食管癌细胞系增殖凋亡及放射敏感性影响研究  

Experimental study of the effect of GOLPH3 on proliferation, apoptosis and radiosensitivity of OE33 esophageal cancer cell line

作　　者：刘杨 蒋月 杨原源 刘晓 徐文才 Liu Yang;Jiang Yue;Yang Yuanyuan;Liu Xiao;Xu Wencai(Department of Radiation Oncology,Affiliated Cancer Hospital of Zhengzhou University/Henan Cancer Hospital,Zhengzhou 450008,China)

机构地区：[1]郑州大学附属肿瘤医院/河南省肿瘤医院放疗科,450008

出　　处：《中华放射肿瘤学杂志》2019年第8期606-609,共4页Chinese Journal of Radiation Oncology

基　　金：河南省科技厅重点科技攻关项目(142102310502);河南省卫生厅普通科技攻关项目(201403026);河南省卫生厅科技攻关指导性计划(201304058).

摘　　要：目的研究GOLPH3对食管癌细胞系增殖凋亡及放射敏感性的影响。方法以qRT-PCR和蛋白印迹法分别检测食管癌细胞系和正常食管上皮细胞GOLPH3 mRNA和蛋白水平,在OE33细胞中转染GOLPH3 siRNA,同时给予照射处理。MTT检测细胞增殖,流式细胞术检测细胞凋亡,细胞克隆实验检测放射敏感性,蛋白印迹法检测活化的Caspase-3(Cleaved Caspase-3)、Bcl-2相关X蛋白(Bax)、活化的Caspase-9(Cleaved Caspase-9)蛋白水平。结果食管癌细胞中GOLPH3 mRNA和蛋白水平明显高于正常食管上皮细胞(P<0.05)。GOLPH3 siRNA可明显下调食管癌OE33细胞中GOLPH3 mRNA和蛋白水平。下调GOLPH3或者照射处理以后的食管癌细胞的增殖活性降低,细胞凋亡率升高,并且细胞中Cleaved Caspase-3、Bax、Cleaved Caspase-9蛋白水平也升高(P<0.05)。下调GOLPH3后的OE33细胞经照射处理以后,细胞增殖活性下降更多,细胞凋亡率和细胞中Cleaved Caspase-3、Bax、Cleaved Caspase-9蛋白水平升高更多(P<0.05)。下调GOLPH3后OE33细胞经照射处理后,细胞放射增敏比为1.673。结论GOLPH3在食管癌细胞中高表达,下调其表达可以增加OE33细胞放射敏感性,诱导细胞凋亡并抑制其增殖。Objective To evaluate the effect of GOLPH3 on the proliferation, apoptosis and radiosensitivity of OE33 esophageal cancer cell line. Methods The expression levels of GOLPH3 mRNA and protein in the esophageal cancer cells and normal esophageal epithelial cells were detected by qRT-PCR and Western blot, respectively. The OE33 esophageal cancer cells were transfected with GOLPH3 siRNA and subject to irradiation treatment simultaneously. The cell proliferation was detected by MTT assay. The cell apoptosis was detected by flow cytometry. The radiosensitivity was assessed by cell cloning test. The expression levels of cleaved Caspase-3, Bax and cleaved Caspase-9 protein levels were quantitatively measured by Western blot. Results The expression levels of GOLPH3 mRNA and protein in the esophageal cancer cells were significantly higher than those in the normal esophageal epithelial cells (both P<0.05). GOLPH3 siRNA could obviously down-regulate the expression levels of GOLPH3 mRNA and protein in the OE33 esophageal cancer cells. The proliferation activity of esophageal cancer cells was decreased, whereas the apoptosis rate was increased and the expression levels of cleaved Caspase-3, Bax and cleaved Caspase-9 were up-regulated after down-regulating the expression of GOLPH3 or irradiation treatment (all P<0.05). After down-regulating the expression of GOLPH3 in the esophageal cancer cells treated with irradiation, the cell proliferation activity was more significantly decreased, whereas the apoptosis rate was elevated and the expression levels of cleaved Caspase-3, Bax and cleaved Caspase-9 were more evidently up-regulated (all P<0.05). In the irradiated OE33 esophageal cancer cells after down-regulating the expression of GOLPH^3, the radiosensitization ratio of the cells was 1.673.Conclusions GOLPH3 is highly expressed in the esophageal cancer cells. Down-regulating the expression of GOLPH3 can increase the radiosensitivity, induce the apoptosis and inhibit the proliferation of OE33 esophageal cancer cells.

关 键 词：食管癌细胞系 凋亡 放射敏感性 GOLPH3基因 

分 类 号：R735.1[医药卫生—肿瘤]