摘要
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 investigate the effects of adenovirus-mediated ING4 gene （Ad-ING4 ） on human prostatic carcinoma cell PC-3 in vitro. Methods After Ad-ING4 was transfected into PC-3 cells, the ING4 gene was transeripted in PC-3 cells confirmed by RT-PCR. The cytotoxicity of Ad-ING4 to PC-3 cells was observed by fluorescent microscopy. The cellular apoptosis of PC-3 cells induced by Ad-ING4 was detected by FCM. The proliferation curve of PC-3 cells was drawn by MTT assay, and the inhibitory rate was calculated. The morphological changes of the nuclei were observed with Hoechst 33258 staining. Results The transcription and expression of ING4 gene in PC-3 cells was detected by RT-PCR. After infected with Ad-ING4 for 72h and 96h, the growth inhibitory rates were 39. 29% and 50% respectively. After infected with Ad-ING4 for 72h, the apoptosis rate of PC-3 cells was 76.19 %. The Hoechst 33258 staining suggested that Ad-ING4 could induce PC- 3 cells to apoptosis, showing the classical morphological changes of the nuclei. Conclusions Ad-ING4 can inhibit the proliferation of PC-3 cells, and induce apoptosis in PC-3 cell line.
出处
《中国校医》
2011年第3期239-240,F0003,共3页
Chinese Journal of School Doctor
基金
2008年国家大学生创新性实验计划项目基金(SG315859)
2009年苏州大学大学生创新性实验计划项目基金(57315518)
