Ad-ING4-IL-24双基因共表达腺病毒对前列腺癌细胞株PC3体内外抑癌增效作用

Enhanced anti-tumor effects of recombinant adenovirus expressing ING4 and IL-24 gene on PC3 prostate cancer cells and the mechanisms in vitro and in vivo

目的 观察肿瘤生长抑制因子4（ING4）和白细胞介素-24（IL-24）双基因共表达腺病毒载体对PC3人前列腺癌细胞体内外抑癌增效作用.方法 应用半定量逆转录-聚合酶链反应（RT-PCR）和Western blot法检测ING4和IL-24在PC3细胞中的表达;噻唑蓝（MTT）比色法和流式细胞术（FCM）检测双基因对PC3细胞的生长抑制和凋亡效应;半定量RT-PCR法和Western blot法检测双基因的表达对PC3细胞中的bcl-2、bax、p53和Caspase-3凋亡相关基因表达的影响.在前列腺癌的裸鼠移植瘤动物模型上,检测Ad-ING4-IL-24对移植瘤生长的抑制作用,并通过免疫组织化学法检测bcl-2、bax、Caspase-3、CD34等相关因子的表达.结果腺病毒介导的ING4和IL-24双基因在PC3细胞中成功表达,对PC3细胞增殖具有抑癌增效功能,可引起p53、bax、Caspase-3基因表达上调和bcl-2基因表达下凋,诱导细胞凋亡.Ad-ING4-IL-24能显著抑制PC3裸鼠前列腺癌移植瘤生长,瘤重的抑制率可达74%,免疫组织化学结果显示Ad-ING4-IL-24能明显上调bax和Caspase-3的表达和下调bcl-2和CD34基因表达.结论 Ad-ING4-IL-24在体内外均可明显抑制PC3细胞的生长,诱导其凋亡,具有抑癌增效功能.其机制可能是通过上凋p53、bax、Caspase-3基因和下凋bcl-2和CD34基因表达水平.

Objective To construct a recombinant adenoviral vector co-expressing inhibitor of growth 4 （ING4） and interleukin-24 （IL-24） （Ad-ING4-IL-24） and study its enhanced anti-tumor effects on PC3 prostate cancer cells and the mechanism.Methods Semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting were used to detect the expression of ING4 and IL-24 in PC3 cells.The growth inhibition and apoptosis effect were measured by methylthiazol tetrazolium （MTT） and flow cytometry （FCM） respectively.semi-quantitative RT-PCR and Western blotting were used to detect the expression of cell apoptosis-related genes （ bcl-2,bax and Caspase-3 ） in PC3 cells.In athymic nude mice bearing PC3 tumors,Ad-ING4-IL-24 was intratumorally injected,and the changes in the tumor growth were observed.The expression of bcl-2,bax,Caspase-3 and CD34 genes was examined by using immumohistochemistry.Results Ad-ING4-IL-24 was proved successfully transcribed and translated into PC3 cells,significantly inhibited growth of PC3 cells,and exerted enhanced anti-tumor effects.Ad-ING4-IL-24 induced apoptosis by up-regulating the expression of p53,hax and Caspase-3 and downregulating the expression of bcl-2.In vivo,intratumoral injection of Ad-ING4-IL-24 suppressed the tumor growth obviously with a inhibition rate of 74%.Immumohistochemistry revealed that the expression of bax and Caspase-3 was up-regulated and that of bcl-2 and CD34 was down-regulated by Ad-ING4-IL-24.Conclusion Ad-ING4-IL-24 could obviously inhibit the growth of PC3 cells,induce apoptosis and exert the enhanced anti-tumor effects s in vitro and in vivo by up-regulating p53,bax,Caspase-3 and down-regulating bcl-2 and CD34.