文档介绍：ppGalNAcT2反义RNA对胃癌细胞SGC7901生物学特性的影响
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作者:金美芳, 邵雪军, 吴士良
【摘要】目的: 研究人胃癌细胞SGC7901中多肽N乙酰氨基半乳糖转移酶2 (ppGalNAcT2) 基因表达与肿瘤转移相关基因MMP2,TGFβ1表达的相关性,以及对细胞增殖和形态生物学特性的影响。方法: 用两个已构建好的反义表达载体转染胃癌细胞SGC7901,通过G418筛选,建立一系列旨在封闭胃癌细胞SGC7901 ppGalNAcT2基因表达的亚细胞克隆。通过共聚焦显微镜,RTPCR检测反义封闭ppGalNAcT2基因RNA表达后,胃癌细胞SGC7901中TGFβ1,MMP2表达水平,细胞增殖以及形态的变化。结果: 反义封闭ppGalNAcT2基因表达后, 胃癌细胞SGC7901 ppGalNAcT2的表达水平明显降低,细胞的形态发生改变,分裂增殖减慢。结果还显示,反义封闭ppGalNAcT2基因表达可使TGFβ1,MMP2基因在mRNA表达水平均增加,提示ppGalNAcT2基因表达可能对胃癌细胞SGC7901浸润转移产生影响。结论: ppGalNAcT2可能在肿瘤的增殖、浸润和转移中发挥作用。
【关键词】 SGC7901细胞; 多肽N
乙酰氨基半乳糖转移酶; 基质金属蛋白酶; 转化生长因子β1
[Abstract] Objective: To study the effect on the cell proliferation and the level of MMP2 and TGFβ1 in the SGC7901 cells caused by antisense blocking ppGalNAcT2 gene expressionMethods: The antisense expressing vectors of two different length ppGalNAcT2 gene segments were constructed and transfected into SGC7901 series of subcellular clones aiming at blocking of ppGalNAcT2 gene expression of SGC7901 cells were established by G418 antisense blocking, the cells were first tested with fluorescent the expression of ppGalNAcT2,MMP2 and TGFβ1 gene cells growth were studied with RT effects of ppGalNAcT2 antisense RNA on the proliferation of SGC7901 cells were tested by MTT. Results: The results showed that ppGalNAcT2 level in SGC7901 cells was obviously redudced and the cell proliferation was slower after antisense indicated that the blocking of ppGalNAcT2 gene expression had influence on SGC7901 cell was also revealed that the blocking of ppGalNAcT2 gene expression could increase the mRNA and protein level of MMP2 and TGF
β1. Conclusion: The result suggests that ppGalNAcT2 has an important role in the cell proliferation, invasion and metastasis of cancer.
[Key words] SGC7901 cell; polypeptide Nacetylgalactosaminyltransferase; matrix metalloprotei