PCNA基因反义RNA表达质粒的构建及对人胃癌裸鼠移植瘤的抑制效应

CONSTRUCTION OF PCNA ANTISENSE RNA EXPRESSION VECTOR AND ITS ANTITUMOR EFFECT ON HUMAN GASTRIC CANCER XENOGRAFTS IN NUDE MICE

为观察ＰＣＮＡ基因反义ＲＮＡ表达质粒对人胃癌裸鼠移植瘤的抑制效应，用ＤＮＡ重组法将人ＰＣ－ＮＡ基因反向克隆到真核细胞表达质粒ｐＤＯＲ－ｎｅｏ中，构建成ＰＣＮＡ基因真核表达质粒ｐＤＲ－ＰＣＮＡ，用ＬｉｐｏｆｅｃｔＡＭＩＮＴＭ介导转染人胃癌细胞系ＳＧＣ－７９０１，并接种于裸鼠背部皮下。经Ｇ４１８筛选获得的转染细胞系ＳＧＣ／ＰＣＮＡ与亲本细胞相比，其生长速度减慢，ＲＮＡ、蛋白质生物合成受到抑制，Ｓ期、Ｇ２／Ｍ期ＤＮＡ含量降低，移植瘤生长速度减慢，瘤体平均直径（０．５４±０．１３）ｃｍ，明显小于对照组（１．５１±０．１１）ｃｍ。结果表明，ＰＣＮＡ基因反义ＲＮＡ表达质粒对胃癌裸鼠移植瘤生长具有明显的抑制作用。

To observe the effect of proliferating cell nuclear antigen(PCNA) antisense RNA on human gastric cancer xenografts in immuno compromized Balb/c mice and their antitumor roles, an eukaryotic expression vector pDR PCNA was successfully constructed with DNA recombinant by inserting the human PCNA gene into an eukaryotic expression pDOR neo in reverse direction. The human gastric cancer cell line SGC 7901 was tansfected with pDR PCNA by lipofect AMINE TM , which was inoculated into the bake of mude mice. After G418 selection, the growth rate of cells, the biosyntheses of proteins and RNA and the DNA contents of transfectants in S phase and G2/M phase were significantly suppressed in comparison with that of the parental cell line. The tumor formation time and growth rate of transplantable tumor in nude mice inoculated with SGC/PCNA greatly reduced in comparison with that in nude mice inoculated with SGC 7901, or SGC/neo. The average diameter (0 54±0 13)cm of transplantable tumor in nude mice inoculated with SGC/PCNA was markedly smaller than that of controls (1 51±0 11)cm. The results demonstrated that PCNA antisense RNA could markedly suppress the growth of human grastric cancer transplantalbe tumor in nude mice.