CAJ | 학술논문

目的比较RNA干扰(RNAi)和伊马替尼(imatinib)杀伤K562细胞的效果.方法设计有效的bcr-abl干扰shRNA序列,利用基因工程技术插入到干扰载体构建RNAi质粒.测序鉴定正确的RNAi质粒转染K562细胞,48 h后荧光显微镜观察转染效率.RT-PCR技术检测K562细胞中hcr-ahl表达水平.同时,伊马替尼作用K562细胞.利用凋亡分析、MTT增生实验和磷酸化的酪氨酸蛋白含量检测来评估RNAi和伊马替尼作用效果.结果与伊马替尼一样,RNAi使K562细胞凋亡增强,增生减少,磷酸化的酪氨酸蛋白含量减少.结论 RNAi达到伊马替尼杀伤K562细胞的效果,有望在临床用于治疗慢性髓细胞白血病(CML)患者,尤其是那些对伊弓替尼等化疗药物不敏感的CML患者.
목적 비교RNA간우(RNAi)화이마체니(imatinib)살상K562세포적효과.방법 설계유효적bcr-abl간우shRNA서렬,이용기인공정기술삽입도간우재체구건RNAi질립.측서감정정학적RNAi질립전염K562세포,48 h후형광현미경관찰전염효솔.RT-PCR기술검측K562세포중hcr-ahl표체수평.동시,이마체니작용K562세포.이용조망분석、MTT증생실험화린산화적락안산단백함량검측래평고RNAi화이마체니작용효과.결과 여이마체니일양,RNAi사K562세포조망증강,증생감소,린산화적락안산단백함량감소.결론 RNAi체도이마체니살상K562세포적효과,유망재림상용우치료만성수세포백혈병(CML)환자,우기시나사대이궁체니등화료약물불민감적CML환자.
Objective To compare RNA interference (RNAi) with imatinib in killing K562 cells. Methods Design effective shRNA sequences special for bcr-abl silencing and insert them into the eukaryotic expression vector for RNAi by gene engineering. The recombinant plasmi(ts were then transfected into K562 cells. 48 hours later, the efficiency of transfection was identified by fluorescent microscope, bcr-abl mRNA level was detected by RT-PCR. Another group of K562 cells were treated respectively by imatinib with different concentration. All groups of K562 cells were finally analyzed in apoptosis, cell proliferation and phosphotyrosine-containing proteins. Results Both RNAi and imatinib induced apoptosis, decreased proliferation and reduced phosphotyrosine-containing proteins. Conclusion BNAi can kill K562 cells successfully as imatinib, and it may be a promising way to treat CML patients in clinic, especially for those who fail in imatinib or other chemotherapy.