ppGalNAcT2反义RNA对胃癌细胞SGC7901生物学特性影响.docVIP

ppGalNAcT2反义RNA对胃癌细胞SGC7901生物学特性影响 　 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 cells.A series of subcellular clones aiming at blocking of ppGalNAcT2 gene expression of SGC7901 cells were established by G418 selection.After antisense blocking, the cells were first tested with fluorescent microscopy.Then the expression of ppGalNAcT2，MMP2 and TGFβ1 gene cells growth were studied with RTPCR.The 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 blocking.It indicated that the blocking of ppGalNAcT2 gene expression had influence on SGC7901 cell proliferation.It 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 metalloproteinase; transforming growth factorβ1