目的:构建人抑癌基因VHL的真核表达载体,并验证其对肿瘤细胞生长的影响。方法:采用PCR技术从人乳腺文库中扩增人VHL基因,将其克隆到p XJ-40-myc载体中,酶切和测序验证后转染人胚肾293T细胞,通过蛋白免疫印迹鉴定其表达;转染人乳腺癌ZR75-1细胞和肝癌Hep G2细胞,通过CCK8法测定细胞生长曲线。结果:从人乳腺文库中扩增得到约650 bp的DNA片段,并克隆至p XJ-40-myc载体上,且测序与目的序列完全一致;转染人胚肾293T细胞后,蛋白免疫印迹检测到相对分子质量为26×103的目的基因表达产物;细胞生长曲线显示,转染myc-VHL的乳腺癌、肝癌细胞较空载体细胞生长慢。结论:构建了myc-VHL真核表达载体,myc-VHL抑制癌细胞生长,为进一步研究VHL在肿瘤发生发展中的功能奠定了基础。 Objective: To construct eukaryotic expression vector of human tumor suppressor gene VHL and verify its effect on tumor cell growth. METHODS: Human VHL gene was amplified from human breast library by PCR and cloned into pXJ-40-myc vector. After verified by restriction enzyme digestion and sequencing, the human embryonic kidney 293T cells were transfected and their expression was identified by Western blot. Transfected human breast cancer ZR75-1 cells and hepatoma Hep G2 cells, cell growth curve was determined by CCK8 method. RESULTS: Approximately 650 bp DNA fragment was amplified from human mammary gland library and cloned into pXJ-40-myc vector. The sequencing was completely consistent with the target sequence. After transfection of human embryonic kidney 293T cells, Western blot analysis was performed. The target gene expression product with a relative molecular weight of 26×103 was obtained; the cell growth curve showed that the breast cancer and hepatoma cells transfected with myc-VHL grew slower than the empty vector cells. Conclusion: The myc-VHL eukaryotic expression vector was constructed and myc-VHL inhibited the growth of cancer cells, which laid the foundation for further research on the function of VHL in tumor development.