Ovol2对食管鳞癌细胞增殖、迁移能力的影响和机制研究  

Effect of Ovol2 on the proliferation and migration of esophageal squamous cell carcinoma cells and its related mechanism

作　　者：韩晖琼 王磊[1,2] 秦艳茹[1,2] HAN Hui-qiong;WANG Lei;QIN Yan-ru(Department of Oncology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan,CHINA;State Key Laboratory of Esophageal Cancer Prevention and Control,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan,CHINA)

机构地区：[1]郑州大学第一附属医院肿瘤科,河南郑州450052 [2]郑州大学第一附属医院省部共建食管癌防治国家重点实验室,河南郑州450052

出　　处：《海南医学》2024年第15期2129-2134,共6页Hainan Medical Journal

基　　金：国家自然科学基金面上项目(编号:82273381)。

摘　　要：目的初步探索转录因子Ovo样锌指2(Ovol2)对食管鳞状细胞癌(ESCC)增殖、迁移能力的影响和相关机制。方法体外培养食管鳞癌细胞系HKESC1、EC109、EC9706、KYSE30、KYSE140、KYSE180、KYSE410、KYSE510、KYSE520和正常食管上皮细胞系NE1,用实时荧光定量PCR(RT-qPCR)检测上述细胞系中Ovol2的m RNA表达水平,采用Western blot检测上述细胞Ovol2蛋白表达水平、EC9706和TE1稳定株对照组(DMSO处理)和实验组(Doxycycline处理)Ovol2蛋白表达水平及EC9706和TE1细胞空白对照组(Blank)、稳定株对照组(DMSO处理)和稳定株实验组(Doxycycline处理)中上皮间充质转化(EMT)相关基因的蛋白表达水平;采用慢病毒感染的方法构建Ovol2诱导过表达稳定株,1μg/mL的Doxycycline处理稳定株诱导Ovol2表达,1DMSO处理稳定株作为对照;平板克隆形成实验、CCK-8细胞增殖实验检测细胞增殖能力;划痕愈合实验、Transwell细胞迁移实验检测食管癌细胞迁移能力。结果与正常食管上皮细胞系NE1相比,Ovol2在食管癌细胞系中蛋白表达水平明显降低,且Ovol2在不同食管癌细胞系中的表达存在明显差异,差异均有统计学意义(P<0.05);Ovol2在食管癌细胞系和NE1中的m RNA表达水平差异有统计学意义(P<0.05)。EC9706和TE1细胞实验组的克隆形成数量较对照组减少,其中EC9706对照组为(246.33±11.50)个,EC9706实验组为(83.00±7.55)个,TE1对照组为(170.00±9.00)个,TE1实验组为(20.33±3.06)个,两个细胞系过表达组和对照组的差异均有统计学意义(P<0.05);EC9706和TE1实验组的增殖速率较对照组减慢,差异均有统计学意义(P<0.05)。EC9706和TE1细胞实验组较对照组穿过Transwell小室的细胞数量减少,其中EC9706对照组为(150.00±20.27)个,EC9706实验组为(25.00±7.00)个;TE1对照组为(180.00±10.40)个,TE1实验组为(33.00±6.70)个,以上差异均有统计学意义(P<0.05);EC9706细胞实验组在同样时间内划痕愈合范围较对照组缩小,差异有�Objective To investigate the effect of transcription factor Ovol2 on the proliferation and migration of esophageal squamous cell carcinoma(ESCC)and its related mechanisms.Methods Esophageal squamous cell lines HKESC1,EC109,EC9706,KYSE30,KYSE140,KYSE180,KYSE410,KYSE510,KYSE520,and normal esophageal epithelial cell lines NE1 were cultured in vitro,and the mRNA expression level of Ovol2 in the above cell lines was de-tected by real-time quantitative PCR(RT-qPCR).The expression levels of Ovol2 protein,the expression levels of Ovol2 protein in the control group(DMSO treatment)and experimental group(Doxycycline treatment)of the stable strains EC9706 and TE1 cells,and the protein expression levels of genes related to epithelial mesenchymal transformation(EMT)in the blank control group(Blank),the control group(DMSO treatment),and the experimental group(Doxycy-cline treatment)of the EC9706 and TE1 cells were detected by western blot.The stable strain of Ovol2 induced overex-pression was constructed by lentivirus infection,the stable strain treated with 1μg/ml of doxycycline induced the expres-sion of Ovol2,and the stable strain treated with 1‰DMSO was used as the control.Plate cloning assay and CCK8 cell proliferation assay were used to detect cell proliferation ability.Scratch healing assay and Transwell cell migration assay were used to detect the migration ability of esophageal cancer cells.Results Compared with the normal esophageal epi-thelial cell line NE1,the protein expression level of Ovol2 in esophageal cancer cell lines was significantly reduced,and there were significant differences in the expression of Ovol2 in different esophageal cancer cell lines(P<0.05).There were significant differences in the mRNA expression levels of Ovol2 in esophageal cancer cell lines and NE1(P<0.05).The number of clones formed in the EC9706 and TE1 cell experimental groups were reduced,and the number was 246.33±11.50 in EC9706 control group,83.00±7.55 in EC9706 experimental group,170.00±9.00 in TE1 control group,and 20.33±3.

关 键 词：食管鳞癌 Ovo样锌指2 上皮间充质转化 细胞增殖 细胞迁移 

分 类 号：R735.1[医药卫生—肿瘤]