TGM2对胃癌细胞BGC-823增殖、迁移和侵袭能力的影响  

The Role of TGM2 on Cell Proliferation, Migration and Invasion of BGC-823 Cell in Gastric Cancer

作　　者：雷志杰 张莹[1] 田苗苗[1] 岳俊涛 夏云龙[3] 胡思隽[1] 吴开春[1] LEI Zhi-jie;ZHANG Ying;TIAN Miao-miao;YUE Jun-tao;XIA Yun-long;HU Si-jun;WU Kai-chun(Xijing Hospital of Digestive Disease,State Key Laboratory of Oncology,Fourth Military Medical University,Xi＇an Shaanxi,710032,China;Department of Urology,Xijing Hospital,Fourth Military Medical University,Xi＇an,Shaanxi,710032,China;Department of Cardiology,Xijing Hospital,Fourth Military Medical University,Xi＇an,Shaanxi,710032,China)

机构地区：[1]第四军医大学西京消化病医院.肿瘤生物学国家重点实验室,陕西西安710032 [2]第四军医大学西京医院泌尿外科,陕西西安710032 [3]第四军医大学西京医院心脏内科,陕西西安710032

出　　处：《现代生物医学进展》2018年第10期1816-1822,共7页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81421003;81627807;91542000;81672702;81701750)

摘　　要：目的:转谷氨酰胺酶-2(transglutaminase-2,TGM2)是一种多功能的蛋白,在乳腺癌,胰腺癌,黑色素瘤和前列腺癌等肿瘤中高表达,而且和肿瘤的增殖和转移密切相关。本研究旨在探索TGM2对胃癌细胞BGC-823的增殖、迁移和侵袭能力的影响。方法:q RT-PCR、蛋白质印迹及免疫组化检测TGM2在胃癌细胞及组织中的表达情况;慢病毒转染胃癌细胞BGC-823以干扰TGM2的表达,荧光显微镜下观察转染效率,蛋白质印迹检测TGM2表达干扰效果;CCK-8法检测TGM2下调对BGC-823增殖能力的影响;Transwell法检测TGM2下调对BGC-823的迁移和侵袭能力影响;蛋白质印迹检测TGM2下调对转移和凋亡相关蛋白表达的影响。结果:TGM2在胃癌细胞和组织中均高表达,P<0.01(q RT-PCR),P<0.001(免疫组化);转染慢病毒后,荧光显微镜下观察结果显示转染效率超过90%,蛋白质印迹结果显示实验组sh TGM2-1的干扰效果最好,TGM2明显下调,差异具有统计学意义,P<0.0001;CCK-8结果表明TGM2下调后,从细胞铺板第2d开始,实验组(sh TGM2-1)的细胞增殖倍数明显下降,P<0.05(2d),P<0.01(3d),P<0.001(4d),P<0.01(5d);Transwell结果显示,TGM2下调后,BGC-823的迁移和侵袭能力明显减弱,P<0.001(迁移),P<0.01(侵袭);蛋白质印迹结果显示,TGM2下调后,NF-κB和Bcl-2表达下调,而Bax表达上调,P<0.05(NF-κB)、P<0.001(Bcl-2)和P<0.0001(Bax)。结论:TGM2下调能抑制胃癌细胞BGC-823的增殖、迁移和侵袭能力,并和NF-κB、Bcl-2的下调以及和Bax的表达水平上调有关,为胃癌的临床治疗提供了新靶点。Objective： Transglutaminase-2（TGM2） is a multifunctional protein that is overexpressed in tumors such as breast,pancreatic,melanoma and prostate cancer,and is closely related to tumor proliferation and metastasis.The purpose of this study was to explore the effect of TGM2 on the proliferation,migration and invasion of gastric cancer cell line BGC-823.Methods： The expression of TGM2 in gastric cancer cells and tissues was detected by q RT-PCR,Western blotting and immunohistochemistry.The lentivirus was used to transfect BGC-823 to down-regulate the expression of TGM2 and the transfection efficiency was observed by fluorescence microscopy and Western blotting.The effect of TGM2 down-regulation on the proliferation of BGC-823 cells was detected by CCK-8 assay.The effect of down-regulation of TGM2 on the migration and invasion ability of BGC-823 was detected by Transwell assay.The down-regulation of TGM2 on the metastasis and apoptosis-related protein expression was detected by Western blotting.Results： TGM2 was highly expressed in gastric cancer cells（P〈0.01） and tissues（P〈0.001）.The results of fluorescence mi-croscopy showed that transfection efficiency was more than 90 %.The results of Western blotting indicated that the interference effect of sh TGM2-1 group was best obvious（P〈0.0001）.The results of CCK-8 showed that after TGM2 down-regula-tion,from the first day,cell proliferation in the experimental group（sh TGM2-1） was significantly decreased.P〈0.05（2 d）,P〈0.01（3 d）,P〈0.001（4 d） and P〈0.01（5 d）.Transwell assays showed that the migration（P〈0.001） and invasion（P〈0.01） ability of BGC-823 were sup-pressed after TGM2 down-regulation.Western blotting showed that,expression of NF-κB（P〈0.05） and Bcl-2（P〈0.001） were down-regulated while Bax（P〈0.0001） was up-regulated in BGC-823 when TGM2 was down-regulated.Conclusions： The proliferation,migration and invasion abilities of gastric cancer cell BGC-823 are inhibited significantly afte

关 键 词：胃癌 TGM2 细胞增殖 肿瘤转移 

分 类 号：R-33[医药卫生] R735.2