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目的研究腺病毒介导的ING4基因(简称Ad-ING4)体外对人前列腺癌细胞PC-3的生长抑制作用。方法将携有绿色荧光蛋白(GFP)的腺病毒空载体Ad(简称Ad-GFP)及Ad-ING4分别感染PC-3细胞,RT-PCR检测ING4基因的转录;荧光显微镜观察Ad-ING4对PC-3细胞的细胞毒作用;用MTT比色法绘制细胞生长曲线,计算生长抑制率;流式细胞仪(FCM)检测Ad-ING4对细胞凋亡的影响;Hoechst 33258核荧光染色检测细胞凋亡的核形态学变化。结果 RT-PCR显示Ad-ING4能在PC-3细胞内转录;MTT结果提示Ad-ING4感染72 h和96 h后对细胞生长抑制率达39.29%和50.00%;FCM检测Ad-ING4感染细胞72 h后凋亡率为76.19%;核荧光染色结果进一步证明Ad-ING4对PC-3细胞可呈现典型的细胞凋亡核形态学改变。结论 Ad-ING4能够明显抑制PC-3细胞的生长,并诱导细胞凋亡。
Objective To study the inhibitory effect of adenovirus-mediated ING4 gene (Ad-ING4) on the growth of human prostate cancer cell line PC-3 in vitro. METHODS: PC-3 cells were infected with adenovirus vector Ad-GFP and Ad-ING4 carrying green fluorescent protein (GFP), respectively. The transcription of ING4 gene was detected by RT-PCR. The cytotoxicity of Ad-ING4 was detected by flow cytometry (FCM). Apoptosis was detected by Hoechst 33258 nuclear fluorescent staining Nuclear morphological changes. The results of RT-PCR showed that Ad-ING4 could transcribe in PC-3 cells. MTT assay showed that the growth inhibition rates of Ad-ING4 cells were 39.29% and 50.00% at 72 and 96 h, respectively. h apoptosis rate was 76.19%. The result of nuclear fluorescence staining further proved that Ad-ING4 could display typical apoptotic nuclear morphological changes in PC-3 cells. Conclusion Ad-ING4 can significantly inhibit the growth of PC-3 cells and induce apoptosis.