鸟氨酸脱羧酶基因反义RNA对前列腺癌细胞生长的抑制作用

为研究鸟氨酸脱羧酶 (ODC)基因反义RNA对前列腺癌细胞的生长抑制作用 ,将表达ODC第 3外显子反义RNA的重组腺病毒rAd ODC Ex3as分别感染前列腺癌细胞株PC 3和LNCap .通过MTT法观察其对前列腺癌细胞增殖的影响 ,并确定不同细胞合适的感染滴度 ,再采用Western印迹和流式细胞术检测rAd ODC Ex3as对细胞中ODC表达的抑制作用、对细胞周期和凋亡的影响以及与CDK抑制物p2 1的关系 .实验显示 ,rAd ODC Ex3as分别以 5 0MOI、2 5MOI感染PC 3和LNCap细胞可明显抑制其生长增殖 ,而不引起细胞毒性作用 ;其对两种细胞中ODC表达的抑制作用分别为4 5 %和 5 9% .流式细胞DNA含量分析证实 ,rAd ODC Ex3as可引起PC 3和LNCap细胞周期G1期阻滞 ,但并未引起凋亡 .通过Western印迹发现 ,细胞中ODC表达的降低可诱导p2 1蛋白的过表达 .结果表明 ,rAd ODC Ex3as在体外能有效地干扰ODC基因的表达 ,并通过诱导p2 1的过表达使其细胞周期停于G1期 ,从而抑制前列腺癌细胞PC 3和LNCap的增殖 ,为其进一步基因治疗的研究打下基础 .

Inhibition of Prostate Cancer Cells with Antisense RNA of Ornithine Decarboxylase Gene

To investigate the effect of antisense RNA of the third extron of ornithine decarboxylase ( ODC) gene on prostate cancer cells PC 3 and LNCap, both cells were infected with rAd ODC/Ex3as. MTT assay showed that rAd ODC/Ex3as could significantly inhibit the growth of PC 3 and LNCap cells by 50 MOI and 25 MOI without cell toxicity. Using Western blotting, it was found that 45% and 59% of ODC protein expression were inhibited in PC 3 and LNCap cells, and the decreased ODC expression could further induce the overexpression of CDKi p21. Cell flow cytometry analysis showed that rAd ODC/Ex3as could lead the infected cell to arrest at G1 phase. rAd ODC/Ex3as could interfere with ODC expression, induce G 1 arrest by p21 overexpression and inhibit growth of PC 3 and LNCap cells.