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目的探索利用抗慢性髓细胞性白血病BCR-ABL mRNA的核酶表达载体转染K562细胞诱导细胞凋亡的效应。方法应用X-tremeGENEQ2介导pAVGS4转染K562细胞,设置空载体作为对照,在转染后的24、48、72和96h分别用AnnexinⅤ+PI双染色、TUNEL等法通过流式细胞仪检测K562细胞凋亡率的变化,在转染后72h利用电子显微镜观察细胞形态学变化。结果pAVGS4转染K562细胞72h,电镜显示实验组可见大量典型的凋亡细胞,而对照组凋亡细胞少见;AnnexinⅤ+PI和TUNEL法通过流式细胞仪检测均发现在转染后24h,实验组和对照组K562细胞的凋亡率无明显差别(P>0.05),随着转染后时间的推移实验组细胞凋亡率进行性增加,而空载体对照组的凋亡率变化不大。结论pAVGS4可有效诱导K562细胞发生凋亡,核酶M1RNA有望成为分子靶向治疗有用的工具之一。
Objective To explore the effect of transfecting K562 cells with ribozyme expressing vector of BCR-ABL mRNA against chronic myelogenous leukemia to induce apoptosis. Methods K562 cells were transfected with pAVGS4 by X-tremeGENEQ2, and empty vector was used as a control. AnnexinⅤ + PI double staining was performed at 24, 48, 72 and 96h after transfection. K562 cells were detected by flow cytometry Apoptosis rate changes, 72h after transfection cells using electron microscopy morphological changes. Results After transfected with pAVGS4 for 72 hours, the apoptotic cells in the control group were rare, and the apoptotic cells in the control group were rare. Electron microscopy showed that apoptotic cells were rare in the control group. Flow cytometry was used to detect the expression of AnnexinⅤ + PI and TUNEL. There was no significant difference in the apoptosis rate of K562 cells between the two groups (P> 0.05). The apoptotic rate of K562 cells increased progressively with the passage of time, but the apoptosis rate of the empty vector control group did not change much. Conclusion pAVGS4 can effectively induce the apoptosis of K562 cells, and ribozyme M1RNA is expected to be one of the useful tools for molecular targeted therapy.