p16基因转染对人胰腺癌细胞系生长抑制作用的研究

The growth inhibitory effects by transfection of p16 gene on human pancreatic cancer cell line

目的 探讨p16基因对人胰腺癌细胞系JF30 5的生长抑制作用。 方法 用脂质体介导法将外源野生型p16基因转染不表达p16的人胰腺癌细胞系JF30 5 ,对转染后细胞DNA、RNA和蛋白进行分析并观察其生物学特性变化。 结果 外源野生型p16基因已整合入细胞并获稳定表达 ,表达有外源p16基因的细胞生长速度及集落形成率均有部分抑制。细胞周期分析可见G1期增加 ,S期下降。电镜观察超微结构出现生长抑制特征。 结论 p16基因可作为胰腺癌基因治疗目的基因之一。

Objective[WT5”BZ] To elvate the growth inhibitory effects by transfection of p16 gene on human pancreatic cancer cell line JF305. [WT5”HZ]Methods[WT5”BZ] Recombinant eukaryotic expression vector pDOR p16 containing exogenous human wt p16 cDNA and vector containing neomycin resistance gene only were introduced by lipofectamine mediated gene transfection into JF305 cell line which did not express endogenous p16. By using PCR amplification, in situ hybridization, and immunocytochemistry, the clones obtained were detected for efficiency of transfection and effect of vector expression and observed for the changes of their biologic characteristics. [WT5”HZ]Results[WT5”BZ] Exogenous wt p16 was successfully transferred into JF305 cells and obtained permanent expression. The growth rate of these transfected JF305 cells in regular medium and soft agar was inhibited. The percentage of phage G 1 cells increased and that of phage S cells decreased by analysing cell cycle. The ultrastructural changes of the cells observed under electronmicroscope revealed growth retardation. [WT5”HZ]Conclusion[WT5”BZ] p16 is a candidate for cancer gene replacement therapy of human pancreatic cancer. [WT5”HZ]