增加B细胞异位基因2蛋白表达抑制结肠癌细胞的增殖与转移  

Enhancement of B-cell translocation gene-2 inhibits proliferation and metastasis of colon cancer cells

作　　者：姜忠敏[1] 张亮[1] 张立东[1] 王守峰[1] 郑末[1] 李艳霞[2] 刘晓智[2] 

机构地区：[1]天津市第五中心医院病理科,300450 [2]天津市第五中心医院中心实验室

出　　处：《中华肿瘤杂志》2015年第5期330-335,共6页Chinese Journal of Oncology

基　　金：天津市滨海新区卫生局科技项目（2012BWKY002、2013BWKY001）

摘　　要：目的通过增加B细胞异位基因2（BTG2）蛋白表达水平的方法，抑制结肠癌细胞的增殖和转移。方法采用Western blot法检测正常人肠上皮细胞HIEC和结肠癌细胞SW620、HT-29、LS174T中BTG2蛋白的表达水平。采用免疫组织化学方法检测40例正常结肠上皮、40例结肠腺瘤和40例结肠癌组织中BTG2蛋白的表达情况。将含BTG2基因全长序列的质粒转染结肠癌SW620细胞，采用四甲基偶氮唑蓝（MTr）法绘制细胞生长曲线，免疫荧光方法检测Ki-67蛋白表达，划痕实验和Transwell双室培养体系检测结肠癌SW620细胞的迁移能力，鼠尾胶Matrigel 3D培养体系检测SW620细胞伪足生长情况。结果Western blot检测结果显示，正常人肠上皮细胞HIEC、结肠癌细胞SW620、HT-29和LS174T中BTG2蛋白相对表达水平分别为0．83±0．12、0．18±0．04、0．20±0．05和0．36±0．07。免疫组织化学方法检测结果显示，BTG2蛋白在正常结肠组织、结肠腺瘤组织和结肠癌组织中的阳性表达率分别为82．5％（33／40）、77．5％（31／40）和17．5％（7／40），组间差异有统计学意义（P〈0．05）。免疫荧光方法检测结果显示，对照组、空载质粒组和BTG2转染组中Ki-67阳性率分别为（76．2±8．0）％、（81．4±9．7）％和（50．1±7．1）％，组间差异有统计学意义（P〈0．05）。划痕实验显示，对照组、空载质粒组和BTG2转染组SW620细胞两侧间的距离分别为（79．27±11．24）μm、（80．65±12．17）μm和（124．77±19．63）μm，组间差异有统计学意义（P〈0．05）。Transwell实验结果显示，对照组、空载质粒组和BTG2转染组SW620细胞的迁移率分别为（78．5±13．1）％、（73．2±12．9）％和（47．4±9．1）％，组间差异有统计学意义（P〈0．05）。经BTG2基因转染的SW620细胞克隆球伪足数量减少，延展性较差。结论BTG2基因参与结肠癌细胞的增殖和转移，有效恢�Objective To inhibit the proliferation and metastasis of colon cancer cells by increasing the expression level of B-cell translocation gene-2 （BTG2）. Methods Western blot assay was used to detect the expression level of BTG2 protein in the normal intestinal epithelial HIEC cells and three colon cancer cell lines SW620, HT-29 and LS174T. The expression of BTG2 protein in normal colonic epithelial tissue, colon adenoma and colon cancer tissue was detected by immunohistochemistry. The plasmid with BTG2 gene full-length sequence was transfected into colon cancer SW620 ceils, and the expression of BTG2 protein was detected by Western blot. The cell growth curve was drawn by MTF test. The Ki-67-positive rate was calculated using immunofluorescence staining. The cell migration of colon cancer cells was detected by scratch test and Transwell double chamber culture system, and the pseudopodia growth of tumor cells was detected by Matrigel 3D culture system. Results Western blot results showed that BTG2 relative expression levels were 0.83±0.12,0.18±0.04, 0.20±0.05 and 0.36±0.07 in normal human intestinal epithelial cells HIEC, and human colon cancer cell line SW620, HT-29 and LS174T, respectively. The results of immunohistochemistry showed that the positive expression of BTG2 protein in normal colorectal tissue, colorectal adenoma and colorectal carcinoma tissues were 82.5% （33/40）, 77.5% （31/40） and 17.5% （7/40）,respectively, with a significant difference between two groups （P〈0.05）. Immunofluorescence results showed that the positive rate of Ki-67 in the control group, empty vector group and BTG2 transfection group was （76.2± 8.0） %, （81.4±9.7） % and （ 50.1±7.1 ） %, respectively, showing a significant difference between two groups （P〈 0.05 ）. The scratch test results showed that in the control group, empty vector group and BTG2 transfection group , the distance of SW620 cells between two sides was （ 79.27 ± 11.24） μm, （ 80.65 ± 12.17） μm and （124.77±19.63）

关 键 词：结肠肿瘤 B细胞异位基因2 细胞增殖 细胞迁移 肿瘤转移 

分 类 号：R735.35[医药卫生—肿瘤]