rAd-ODC/Ex3as对前列腺癌PC-3细胞抑制作用的研究

目的:探讨重组腺病毒rAd鄄ODC/Ex3as对前列腺癌细胞的体外抑制作用及对前列腺癌的作用机制。方法:利用报告基因GFP测定病毒感染效率,用WesternBlot检测rAd鄄ODC/Ex3as是否抑制鸟氨酸脱羧酶(ODC)基因的表达,采用MTT法、Matrigeal侵袭实验观察rAd鄄ODC/Ex3as对前列腺癌细胞PC鄄3恶性表型的影响,并用流式细胞仪检测细胞周期和凋亡。结果:当MOI为50时,腺病毒感染PC鄄3细胞的效率约为70%,WesternBlot证实rAd鄄ODC/Ex3as可抑制ODC基因的表达。rAd鄄ODC/Ex3as以20MOI感染PC鄄3细胞可明显抑制其生长增殖和侵袭性穴P<0.01雪,流式细胞仪检测结果证实rAd鄄ODC/Ex3as可引起PC鄄3细胞G1期阻滞,但并未引起凋亡。结论:rAd鄄ODC/Ex3as在体外能有效地干扰ODC基因的表达,并可抑制前列腺癌细胞PC鄄3的生长与侵袭,诱导其发生G1期阻滞,有望成为治疗前列腺癌的基因药物。

Inhibition of prostate cancer cell PC-3 by rAd-ODC/Ex3as

Objective: To study the effects of rAd-ODC/Ex3as on prostate cancer cells in vitro and explore its effecting mechanism. Methods: The infection rate of rAd-ODC/Ex3as was measured with the aid of GFP expression. Western Blot technique was used to observe the inhibition of ODC in infected tumor cells. The malignant phenotype of PC-3 cells was assessed by growth curve and Matrigeal invasion assay respectively, and flow cytometry was used to analyze cell cycle progress and apoptosis. Results: Approximate 70% of PC-3 cells were infected with rAd-ODC/Ex3as when MOI reached 50. The expression of ODC was inhibited in the infected tumor cells. rAd-ODC/Ex3as could inhibit cell growth and invasive ability at 20 MOI. Meanwhile, flow cytometry proved that rAd-ODC/Ex3as led the infected cells to arrest at G1 phase but had no effect on apoptosis. Conclusion: rAd-ODC/Ex3as can suppress the proliferation and invasive ability of prostate cancer cells, and induce G1 arrest. It may be one of the promising medicines for antisense gene therapy in prostate cancer.