表达反义c-myc RNA的重组逆转录病毒载体对人胰腺癌细胞恶性表型的逆转作用

eversal of malignant phenotype of cells of human pancreatic adenocarcinoma cell line by recombinant retroviral vector expressing antisense c-myc

为研究表达反义ｃ－ｍｙｃ ＲＮＡ的逆转录病毒载体抑制靶基因的表达和翻译，使胰腺癌细胞恶性表型逆转的机理，利用ｐＸＴ１逆转录病毒载体构建了一个能表达反义ｃ－ｍｙｃＲＮＡ的质粒，经病毒包装细胞ＰＡ３１７包装后，使之成为具有感染力的重组病毒。用该病毒感染人胰腺癌细胞系ＰＣ－２细胞，经Ｇ４１８筛选得到稳定的转化细胞系。Ｎｏｒｔｈｅｒｎ

A recombinant retroviral vector expressing antisense c-myc was constructed. The re combinant virus DNA was packaged with packaging cell line PA317 cells by the calcium phosphate co precipitation method. The virus supernatant was used to infect human pancreatic carcinoma cell line PC 2. After selection with G418, resistant colonies were obtained. Stable expression of retrovirus in bothPA317 and PC-2 cells were confirmed by Northern blot hybridization- A down-regulation of endogenousc-myc was found in PC-2 cells infected with antisense c-myc construct. It was dem0nstrated that the an tisense c-myc did inhibit the cell growth rate and 3 H-TdR incorporation rate 0f PC-2 cells. The abilityof colony formation in soft agar and tumorogenicity in nude mice of PC-2 cells were significantly sup pressed by the antisense c-myc. The results implicate that recombinant retroviral vector containing anti sense c-myc could inhibit target gene expression and partly reverse malignant phenotype of pancreaticadenocarcinoma cells.