摘要
目的:研究细胞黏附分子1(cell adhesion molecule 1,CADM1)过表达对人胃癌MKN-45细胞增殖和侵袭的影响并探讨其可能的分子机制。方法:采用Western blotting检测3株胃癌细胞系中CADM1蛋白的表达。构建pcDNA-CADM1真核表达载体,并将其转染MKN-45细胞,采用G418筛选稳定表达CADM1的细胞株,利用Western blotting鉴定所筛选的稳定细胞株。采用CCK-8试剂和Boyden小室分析过表达CADM1对MKN-45细胞增殖和侵袭的影响。利用Western blotting检测细胞增殖和侵袭相关蛋白表达。结果:MKN-45细胞中CADM1蛋白的相对表达水平显著低于MKN-28和SGC-7901细胞(P<0.05)。此外,成功构建pcDNA-CADM1真核表达载体,并获得稳定过表达CADM1的MKN-45细胞株。CCK-8结果显示,与未处理组和pcDNA3.1组相比,pcDNA-CADM1组中MKN-45细胞的增殖明显受到抑制(P<0.05)。Boyden小室体外侵袭实验结果显示,与未处理组(101.53±6.89)和pcDNA3.1组(98.77±7.03)相比,pcDNA-CADM1组中MKN-45细胞穿膜的细胞数(52.35±3.89)显著减少(P<0.05)。Western blotting结果显示,与未处理组和pcDNA3.1组相比,pcDNA-CADM1组中p21蛋白表达显著上调,而MMP-2和MMP-9表达显著下调(P<0.05)。结论:CADM1过表达能明显抑制胃癌细胞的增殖和侵袭能力,因而CADM1有望成为胃癌治疗的新靶点。
AIM： To investigate the effects of CADM1 overexpression on proliferation and invasion of human gastric carcinoma cell line MKN -45. METHODS： The protein levels of CADM1 in 3 human gastric carcinoma cell lines were detected by Western blotting. Eukaryotic expression vector pcDNA - CADM1 was constructed and transfected into MKN -45 cells. The MKN -45 cells stably expressing CADM1 were selected by G418 and identified by Western blotting. Furthermore, CCK - 8 assay and Boyden chamber were used to analyze the effects of CADM1 overexpression on the prolife- ration and invasion of gastric carcinoma cells. Western blotting was also utilized to detect the levels of cell proliferation - and invasion- related proteins. RESULTS： Relative level of CADM1 protein in MKN -45 cells was significantly lower than that in MKN - 28 ceils and SGC - 7901 cells. Additionally, eukaryotic expression vector pcDNA - CADM1 was suc- cessfully constructed and MKN - 45 cells stably expressing CADM1 were obtained. Compared with non - treatment and pcDNA3.1 groups, the proliferation of MKN -45 cells was obviously inhibited in pcDNA - CADM1 group. The result of Boyden chamber showed that the migrated cell numbers in pcDNA - CADM1 group （52.35±3.89） were significantly lower than that in untreated group （101.53±6.89） and pcDNA3.1 group （98.77±7.03）. Compared with non -treatment and pcDNA3.1 groups, the protein level of p21 was significantly up -regulated and protein expression of MMP- 2 and MMP-9 was obviously down -regulated. CONCLUSION： Overexpression of CADM1 may markedly inhibit cell proliferation and reduce invasion ability, and thus may be a novel target for treating gastric carcinoma.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第12期2283-2287,共5页
Chinese Journal of Pathophysiology
基金
河南省教育厅科技攻关项目(No.2009B320008)
河南省卫生厅科技攻关课题(No.200804055)
关键词
胃肿瘤
细胞黏附分子1
细胞增殖
细胞侵袭
Stomach neoplasms
Cell adhesion molecule 1
Cell proliferation
Cell invasion
