CAJ | 학술논문

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Western blot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)；MTT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G1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.
목적 이용RNA간우기술관찰고제열휴극단백70 (HSP70)대결장암세포HT29세포적영향.방법 실험분위4조:공백조、공질립조,령외2조시사용이경구건적2충간우HSP70표체적단발잡RNA(shRNA)질립재체,병차장기전염도결장암세포HT29중,분위실험1조(shRNA-1)화실험2조(shRNA-2).이용역전록-취합매련반응(RT-PCR)화Western blot법관찰HT-29세포중HSP70적표체,새서람(MTT)비색법관찰HT29세포적증식,류식세포의검측기세포주기이급조망적변화.결과 여공백조화공질립조비교,이용단발협RNA기술고제HSP70후,실험2조적HT-29세포중HSP70적RNA화단백표체명현하강(P＜0.05)；MTT검측결과제시HT29결장암세포증식명현수도억제(P＜0.01),세포주기검측결과제시G1기세포조체,동시명현증가료조망세포적비례(P＜0.01).결론 HSP70침묵능억제결장암세포HT29세포적증식,촉진조망.
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 (MTT) 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 may be a treatment strategy against colon cancer.