ppGalNAcT2反义RNA对胃癌细胞SGC7901生物学特性的影响

Effects of ppGalNAcT2 antisense RNA on the biological behacior of human gastric cancer SGC7901 cell line

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SGC7901ppGalNAc-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SGC7901pp-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。

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.