肿瘤抑制基因WWOX转染MDA-MB-231细胞系的实验研究

目的探讨肿瘤抑制基因WWOX与乳腺癌疾病相关性，为进一步研究WWOX基因的作用机制及功能奠定基础。方法构建真核表达载体pcDNA4．0／Myc—His-WWOX，并转染至MDA．MB．231细胞中，采用RT-PCR及Westernblot方法，检测WWOXmRNA及融合蛋白在MDA-MB-231细胞中的表达。结果测序结果显示，RT-PCR法获得的WWOX序列与GenBank中报道的一致。真核表达载体pcDNA4．0／Myc．His—WWOX转染MDA—MB-231细胞48h后，从RT-PCR及Westernblot结果中观察到WWOX基因的有效转录。结论成功构建了肿瘤抑制基因WWOX真核表达载体pcDNA4．0／Myc．His—WWOX，为进一步研究WWOX在乳腺癌发生和进展中的作用及其靶向基因治疗奠定了实验基础。

Study on Tumor Suppress Gene WWOX transfected MDA-MB-231 breast cancer cells

Objective To inverstigate the relationship between tumor suppress gene WWOX and breast cancer and to lay the foundation for further study of the mechanism and function of WWOX gene. Methods We transfected MDA-MB-231 cells with the constructed recombinant plasmid pcDNA4.0/Myc-His- WWOX, and observed the expression of WWOX mRNA and WWOX fusion protein by RT-PCR and Western blot. Results The sequence of amplified WWOX gene was verified by DNA sequencing. The expression of WWOX mRNA and WWOX fusion protein were observed in MDA-MB-231 cells 48 h after transfection with recombinant plasmid pcDNA4.0/Myc-His-WWOX with RT-PCR and Western blot. Conclusion A eukaryotic expression vector pcDNA4.0/Myc-His-WWOX was successfully constructed, which provided a experimental basis for investigating effects of WWOX in the occurrence and development of breast cancer and targeted gene therapy.