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.%ATM; 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 CADM 1 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 cells and SGC - 7901 cells. Additionally, eukaryotic expression vector pcDNA - CADM1 was successfully 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 -rn9 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 .
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