短发卡RNA敲除热休克蛋白70对结肠癌细胞HT29的影响

Short hairpin RNA-mediated gene silencing of heat shock protein 70 inhibits human colon cancer growth

目的利用RNA干扰技术观察敲除热休克蛋白70（HSP70）对结肠癌细胞HT29细胞的影响。方法实验分为4组：空白组、空质粒组，另外2组是使用已经构建的2种干扰HSP70表达的短发卡RNA（shRNA）质粒载体，并且将其转染到结肠癌细胞HT29中，分为实验1组（shRNA-1）和实验2组（shRNA-2）。利用逆转录-聚合酶链反应（RT-PCR）和Westernblot法观察HT-29细胞中HSP70的表达，噻唑蓝（MTT）比色法观察HT29细胞的增殖，流式细胞仪检测其细胞周期以及凋亡的变化。结果与空白组和空质粒组比较，利用单发夹RNA技术敲除HSP70后，实验2组的HT-29细胞中HSP70的RNA和蛋白表达明显下降（P〈0．05）；MTTT检测结果提示HT29结肠癌细胞增殖明显受到抑制（P〈0．01），细胞周期检测结果提示G，期细胞阻滞，同时明显增加了凋亡细胞的比例（P〈0．01）。结论HSP70沉默能抑制结肠癌细胞HT29细胞的增殖，促进凋亡。

Objective To investigate the effect of short hairpin RNA （shRNA） against heat shock protein 70 （HSP70） on HT29 cells. Methods Two shRNA plasmid vectors against HSP70 were constructed and stably transfected into HT29 cells to determine their effects on proliferation, cell cycle distribution and apoptosis of HT29 cells in vitro. Results The results of methyl thiazol tetrazolium （MTr） showed that HSP70 silencing efficiently inhibited growth of HT29 cells in culture （ P 〈 0.05 ） and induced cell cycle arrest at G1 phase （P 〈 0. 01 ） as well as increased apoptosis significantly （ P 〈 0. 01 ）. Conclusion Specific HSP70 shRNA silencing could inhibit colon cancer growth and promote apoptosis, which implied that HSP70 silencing hiay be a treatment strategy against colon cancer.