高迁移率族蛋白B1促进结肠癌细胞增殖、迁移与侵袭的机制探讨  

High mobility group box 1 protein promotes proliferation, migration and invasion of colon cancer cells

作　　者：王思萱 龙露叶 方雪娇 徐建昕 钱诗菡 吕建新[1] WANG Si-xuan;LONG Lu-ye;FANG Xue-jiao;XU Jian-xin;QIAN Shi-han;Lü Jian-xin(Zhejiang Provincial Key Laboratory of Medical Genetics,School of Laboratory Medicine and Life Science,Wenzhou Medical University,Wenzhou 325035,China,E-mail:[e-mail])

机构地区：[1]温州医科大学检验医学院生命科学学院浙江省医学遗传学重点实验室

出　　处：《中国病理生理杂志》2019年第11期1966-1973,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81170257);浙江省自然科学基金资助项目(No.Y2110513)

摘　　要：目的:探讨高迁移率族蛋白B1(HMGB1)在结肠癌细胞中的表达和对结肠癌细胞增殖、迁移和侵袭的影响。方法:采用qPCR与Western blot法检测人结肠癌SW620细胞与正常结肠上皮细胞FHC中HMGB1的mRNA和蛋白表达。通过Lipofectamine 3000转染质粒构建稳定下调HMGB1表达(shHMGB1)的SW620细胞,同时设置阴性对照(shNC)细胞和空白对照(blank)细胞;采用CCK-8、平板集落形成实验和Transwell实验检测细胞的增殖、迁移及侵袭能力。采用Western blot检测HMGB1表达下调对p-ERK、ERK、c-Myc、基质金属蛋白酶(MMP)-2/9、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、Bcl-2及Bax的蛋白表达水平的影响。结果:SW620细胞中HMGB1在mRNA及蛋白水平表达均高于正常结肠上皮细胞FHC(P<0.05)。经qPCR与Western blot验证,HMGB1低表达细胞构建成功。与blank组和shNC组相比,shHMGB1组细胞增殖、迁移和侵袭能力受到显著抑制(P<0.01)。Western blot结果表明,同blank组和shNC组比较,shHMGB1组细胞的MMP-2、MMP-9、N-cadherin、c-Myc、Bcl-2及ERK磷酸化蛋白表达显著下调,Bax蛋白表达显著上调(P<0.05)。结论:HMGB1可促进结肠癌SW620细胞的增殖、迁移和侵袭能力,ERK/c-Myc信号通路参与了这一过程。AIM: To investigate the expression of high mobility group box 1 protein(HMGB1) in colon can-cer cells, and to determine its regulatory roles in colon cancer cell proliferation, migration and invasion. METHODS: qPCR and Western blot were used to quantify the mRNA and protein expression levels of HMGB1 in human colon cancer SW620 cells and normal colonic epithelial FHC cells. HMGB1 shRNA was transfected into the SW620 cells to establish the stable HMGB1-downregulating colon cancer cells(shHMGB1 group), and negative control(shNC) group and blank control(blank) group were also set up. The proliferation, migration and invasion of the cells were determined by CCK-8 assay, colony formation experiment and Transwell chamber assays. Western blot was used to determine the protein levels of p-ERK, ERK, c-Myc, matrix metalloproteinase(MMP)-2/9, E-cadherin, N-cadherin, Bcl-2 and Bax. RESULTS: Both of the mRNA and protein levels of HMGB1 in colon cancer cells were higher than those in the normal colonic epithelial cells(P<0.05). HMGB1 gene was successfully knocked down in SW620 cells. Compared with blank group and shNC group, the proliferation, migration and invasion abilities of the cells in shHMGB1 group were significantly inhibited(P<0.05). The protein levels of MMP-2, MMP-9, N-cadherin, c-Myc, Bcl-2 and p-ERK were reduced notably, while the expression of Bax protein was increased(P<0.05) in shHMGB1 group compared with shNC group and blank group.CONCLUSION: HMGB1 effectively promotes the proliferation, migration and invasion of colon cancer cells through ERK/c-Myc signaling pathway.

关 键 词：高迁移率族蛋白B1 结肠癌 ERK/c-Myc信号通路 细胞迁移 细胞侵袭 

分 类 号：R363.2[医药卫生—病理学] R735.3[医药卫生—基础医学]