Bcl-2 shRNA诱导成人T淋巴细胞白血病细胞株Molt4凋亡的研究  

The effect of Bcl-2 shRNA on the apoptosis of Molt4 cells

作　　者：房宝英[1] 何冬梅[1] 张洹[1] 陈丽[1] 

机构地区：[1]暨南大学医学院血液病研究所,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2007年第6期541-545,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省自然科学基金项目(04010446)

摘　　要：目的:探讨以Pgenesil-1质粒为载体表达的Bcl-2短发夹样RNA(short hairpin,shRNA)诱导Molt4成人T淋巴细胞白血病细胞株凋亡的作用。方法:采用脂质体介导的转染方法将表达Bcl-2 shRNA的两个RNA干扰质粒(Bcl-2 shRNA1和Bcl-2 shRNA2)、阴性组(针对非目的基因所构建的shRNA表达载体)、空白组及单纯脂质体组分别转染Molt4细胞。采用免疫细胞化学法检测转染48 h后的各组细胞Bcl-2蛋白表达水平;MTT法分别检测24,48,72 h各组细胞的增殖活性;用姬姆萨染色和流式细胞仪观察细胞凋亡情况。结果:转染Molt4细胞48 h后,Bcl-2 shRNA1组和Bcl-2 shRNA2组的细胞Bcl-2蛋白表达率均明显下降;与阴性对照shRNA组、空白组和单纯脂质体组相比差异有显著性(P<0.05),Bcl-2 shRNA1组的抑制作用要强于Bcl-2 shRNA2组。MTT测定显示Bcl-2shRNA1组和Bcl-2 shRNA2组细胞增殖活力均明显下降,分别与空白组和单纯脂质体组进行比较差异有显著性(P<0.05);阴性对照shRNA组与单纯脂质体组比较则无明显差异(P>0.05)。Bcl-2 shRNA1和Bcl-2 shRNA2作用于Molt4细胞48 h后,姬姆萨染色可见凋亡细胞。流式细胞仪检测示Bcl-2 shRNA1组和Bcl-2 shRNA2组细胞凋亡率随时间逐渐增高,与空白组和单纯脂质体组进行比较差异有显著性(P<0.05),但Bcl-2 shRNA1组和Bcl-2 shR-NA2组二者间无明显差异(P>0.05)。结论:Bcl-2 shRNA表达载体可促进Molt4细胞的凋亡。Aim ：To explore the apoptosis effects of Bcl-2 short hairpin RNA （shRNA） with Pgenesil- 1 as expressing vector in inducing molt4 cell lines. Methods： Two Bcl-2 shRNAs groups（ Bcl-2 shRNA1 and Bcl-2 shRNA2 ） ,negative group,blank group and liposome group were all transfected into Molt4 ceils with Lipofectamine. Bcl-2 protein expression of every group ceils was detected by immunocyto chemistry at 48 h after they were transfected. The cell proliferation was determined by MTT method at 24 h, 48 h and 72 h. The apoptotic ceils were detected by Giemsa method and flow cytometry. Results： After Molt4 ceils were transfected for 48 h, Bcl-2 protein expression levels of Bcl-2 shRNA1 group and Bcl-2 shRNA2 group ceils obviously decreased. The above two groups had obvious difference as compared with the negative group,blank group and liposome group（P 〈0. 05）. And the suppression effect of Bcl-2 shRNA1 was better than that of Bcl-2 shRNA2. MTT assays indicated that the grow rates of ceils of Bcl-2 shRNA1 group and Bcl-2 shRNA2 were significantly lower than those of blank group and liposome group （P 〈 0.05）. And there was no difference in viable cells levels between negative shRNA group and liposome group （ P 〉 0.05 ）. Apoptotic cells can be seen with Giemsa at 48 h after being transfected. FCM assays indicated that the cell apoptosis rates of Bcl-2 shRNA1 group and Bcl-2 shRNA2 group were higher than that of blank group and liposome group （ P 〈 0.05 ）. And there was no difference between Bcl-2 shRNA1 group and Bcl-2 shRNA2 one （ P 〉 0.05 ）. Conclusion： Bcl-2 shRNA expression vectors can induce Molt4 cells apoptosis.

关 键 词：BCL-2 短发夹样RNA Molt4细胞 cell凋亡 

分 类 号：Q786[生物学—分子生物学]