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目的 研究VP3 对人成骨肉瘤细胞OS 73 2多药耐药株R OS 73 2治疗作用的分子作用机制与途径。方法 VP3 基因经酶切后克隆入质粒 pIRES1,获得重组质粒pIRVP3 。脂质体法 pIRVP3 转染R OS 73 2细胞 ,RT PCR检测VP3 在细胞中的表达。提取瘤细胞基因组DNA进行琼脂糖凝胶电泳。流式细胞仪检测凋亡率。Western blot检测p5 3、bcl 2蛋白表达。结果 VP3 基因已在转录水平表达。电泳见转染组DNA呈梯状条带 ,对照组为单一条带。转染组凋亡率明显高于空白对照组 (P <0 .0 1)。转染组与对照组 p5 3及bcl 2蛋白显色带灰度无明显差异。 结论 VP3 可有效诱导R OS 73 2细胞凋亡 ,其分子作用机制与途径可能不依赖 p5 3及bcl 2作用。
Objective To study the molecular mechanism of action of VP3 on the therapeutic effect of OS-73 2 multi-drug resistant strain R OS 73 2 in human osteosarcoma cells. Methods VP3 gene was cloned into plasmid pIRES1 to obtain recombinant plasmid pIRVP3. The R OS 73 2 cells were transfected with pIRVP3 by liposome method. The expression of VP3 was detected by RT PCR. Tumor cell genomic DNA was extracted for agarose gel electrophoresis. Flow cytometry was used to detect apoptosis rate. Western blot detection p5 3, bcl 2 protein expression. Results The VP3 gene was expressed at the transcriptional level. Electrophoresis see transfection group was DNA ladder, the control group as a single band. Transfection group apoptosis rate was significantly higher than the blank control group (P <0.01). There was no significant difference in gray scale between p5 3 and bcl 2 protein in transfection group and control group. Conclusion VP3 can effectively induce apoptosis of R OS-73 2 cells, and its molecular mechanisms and pathways may not depend on the effects of p5 3 and bcl 2.