摘要
目的研究腺病毒（adenovirus，Ad）介导的ING4基因（Ad—ING4）表达对人乳腺癌的生长抑制作用。方法采用Ad—ING4腺病毒感染人乳腺癌MDA—MB-231细胞，RT-PCR、Westernblot法检测ING4基因在肿瘤细胞中的转录和表达；四唑盐（MTF）法、Hochest33258染色法和流式细胞仪（FCM）检测ING4基因的表达对乳腺癌细胞的生长抑制、周期变化和凋亡效应；半定量RT—PCR检测凋亡相关基因的转录；酶联免疫吸附试验（ELISA）法检测人血管生成素1（Ang一1）的分泌水平；在乳腺癌的裸鼠移植瘤动物模型上检测Ad．ING4对移植瘤生长的抑制作用，并检测Bcl-2，Bax，Caspase-3等细胞凋亡相关因子的表达。结果ING4基因可在MDA—MB-231细胞中成功转录及表达，并对乳腺癌细胞有明显增殖抑制及促凋亡作用，G：／M期阻滞达（24．86±1．24）％；Ad-ING4显著抑制移植瘤生长，瘤重抑制率达49％；免疫组化结果显示Ad-ING4上调Bax和Caspase-3的表达，下调Bcl-2、Survivin和CD34的表达。结论ING4基因在体内外均可明显抑制MDA—MB-231细胞的生长，诱导其凋亡。
Objective To investigate the inhibitory effect of inhibitor 4 of growth（ING4） delivered by adenovirus on human breast carcinoma cell MDA-MB-231. Methods MDA-MB-2S1 human breast carcinoma cells were infected with Ad-ING4. The expression level of ING4 gene was detected by RT-PCR and Western blot; The growth inhibition, cell-cycle alteration, and apoptosis were detected by MTI＇, Flowcytometry and Hochest33258 staining, respectively. RT-PCR was used to detect the transcription of Bax, Bcl-2, Survivin genes; The expression level of Ang-1 gene was detected by ELISA; Ad-ING4 was given intratumorally in athymic nude mice bearing MDA-MB-231 tumors, and then tumor growth was monitored; The expression of Bcl-2, Bax and Caspase-3 was analyzed by immumohistochemistry. Results ING4 was successfully transcribed and translated in MDA-MB-231 cells; Ad-ING4 significantly inhibited the proliferation and induced G2/M phase arrest to（24. 86 ± 1.24）% and cell apoptosis of MDA- MB-231. Intratumoral injection of Ad-ING4 suppressed the tumor growth obviously with a inhibitory rate of 49%; Immumohistochemistry showed that the expression of Bax, Caspase-3 were up-regulated and the expression of Bcl-2, Survivin, CD34 were down-regulated by Ad-ING4. Conclusions Ad-ING4 can inhibit the growth of MDA-MB-231 ceils and induce apoptosis in vitro and in vivo.
出处
《中华普通外科杂志》
CSCD
北大核心
2010年第4期313-317,共5页
Chinese Journal of General Surgery
基金
基金项目：江苏省卫生厅医学科研基金资助项目（H200914）
