In vitro and in vivo inhibitory effect of Ad-ING4 gene on proliferation of human prostate cancer PC-3 cells
10.3321/j.issn:1000-467X.2009.11.006
- VernacularTitle:Ad-ING4对人前列腺癌细胞PC-3体内外抑癌效应的研究
- Author:
Yang HUI-CUI
1
;
Sheng WEI-HUA
;
Xie YU-FENG
;
Miao JING-CHENG
;
Wei WEN-XIANG
;
Yang JI-CHENG
Author Information
1. 苏州大学医学部
- Keywords:
ING4 gene;
adenovirus vector;
PC-3 cells;
prostate carcinoma;
tumor-suppression
- From:Chinese Journal of Cancer
2009;28(11):1149-1157
- CountryChina
- Language:Chinese
-
Abstract:
Background and Objective:Adenovirus vector has been widely used in tumor gene therapy.ING4 is a member of growth inhibiting factors and a potent anti-tumor gene which could induce apoptosis of many tumor cells.This study was to investigate the inhibitory effects of adenovirusmediated ING4(Ad-ING4)gene on the proliferation of human prostate cancer PC-3 cells in vitro and in vivo,and to explore its mechanisms.Methods:Ad-ING4 was obtained by virus-amplification technique.After transfection of purified Ad-ING4 into PC-3 cells,the expression of ING4 was detected by reverse transcription-polymerase chain reaction(RT-PCR);the influence of Ad-ING4 transfection on cell proliferation was evaluated using MTT assay.Cell apoptosis was assessed using Hoechst33258 staining and flow cytometry. RT-PCR was performed to detect the mRNA levels of the transcription of apoptosis-related genes such as bcl-2,bax,p53,and caspase-3.Athymic nude mice bearing PC-3 tumors were intratumorally injected with Ad-ING4 (100 μL,1×10~9 pfu/mL). Tumor growth was recorded.All nude mice were killed at the end of the experiment to observe the growth of xenografts. The expressions of Bcl-2, Bax, Caspase-3, and CD34 proteins in tumor tissues were detected by immunohistochemistry. Results: Human ING4 gene was successfully transcribed in PC-3 cells and induced apoptosis by up-regulating p53,bax,caspase-3 expression and down-regulating bcl-2 expression. Inhibition of cell proliferation was significant in PC-3 cells. Tumor growth was significantly inhibited in the Ad-ING4 group as compared with that in the Ad-GFP group and the PBS group (P＜0.05). The weight inhibitory rate was 37.0% in the Ad-ING4 group. The expressions of Bax and Caspase-3 were up-regulated,and the expressions of Bcl-2 and CD34 were downregulated in the Ad-GFP group. Conclusions: Adenovirus-mediated ING4 gene exhibits anti-tumor ability in human prostate cancer PC-3 cells in vitro and in vivo,and induces apoptosis. This may be related to the upregulations of p53, bax, Caspase-3 and down-regulation of bcl-2.
