EBV-miRNA-BHRF1-1对鼻咽癌细胞p53基因表达的影响

Implication of EBV-miRNA-BHRF1-1 on p53 expression in nasopharyngeal carcinoma cells

目的通过研究EBV-miRNA-BHRF1-1对鼻咽癌细胞p53基因表达的调控作用,探讨其对鼻咽癌患者基因治疗的应用价值。方法利用已构建好的EBV-miRNA-BHRF1-1表达质粒及空质粒载体,分别作为转染BHRF1-1质粒组和空质粒组,设置加入PBS转染的CNE-2细胞株作为对照组(PBS组),转染至鼻咽癌细胞株。采用RT-PCR方法检测鼻咽癌相关基因(p53基因)的mRNA水平,Western检测p53蛋白表达水平,并利用CCK-8比色法检测鼻咽癌细胞的生长情况。结果BHRF1-1组细胞BHRF1-1表达量为(0.98±0.05),高于空质粒组(0.66±0.10)及PBS组(0.65±0.12)(均P<0.01),BHRF1-1组、空质粒组、PBS组的细胞p53 mRNA表达量分别为(0.65±0.07)、(0.98±0.06)、(0.99±0.03),BHRF1-1组均低于空质粒组和PBS组(均P<0.01);BHRF1-1组细胞内p53蛋白表达量低于空质粒组和PBS组。BHRF1-1组的细胞增殖抑制率(14.9%)高于空质粒组(11.2%)和PBS组(2.5%)(均P<0.01)。结论 EBV-miRNA-BHRF1-1能有效的调控p53基因mRNA水平及其蛋白翻译水平,并可能对鼻咽癌细胞株的增殖有一定的抑制作用。

Objective To research the regulation function of EBV-miRNA-BHRF1-1 on p53 ex- pression in nasopharyngeal carcinoma cells, and to explore its application value in the treatment of nasopha- ryngeal carcinoma. Methods Plasmid expressing EBV-miRNA-BHRF1-1 and empty plasmid vector were used to transfect into nasopharyngeal carcinoma cells as BHRF1-1 group and empty plasmid group, and set the CNE-2 cells join in PBS as PBS group（control group）. Then, p53 gene mRNA level was detected by RT-PCR method and p53 protein expression level, by Western method, and the growth of nasopharyngeal carcinoma cells was examined by CCK-8 colorimetric method. Results BHRF1-1 expression in BHRF1-1 group was （0. 98 ± 0. 05 ）, higher than that in empty plasmid group （0. 66 ± 0. 10） and PBS group （0. 65 ± 0. 12） （both P 〈 0. 01 ）. The expression of p53 mRNA in cells in BHRF1-1 group, empty plasmid group, and PBS group were （0. 65 ± 0.07）, （0. 98 ± 0.06）, and （0. 99 ± 0. 03 ）, respectively. The p53 mRNA and p53 protein expression level in BHRF1-1 group were lower than the other two groups （ both P 〈 0.01 ）. The cellular proliferation inhibition rate in BHRF1-1 group was 14. 9%, higher than that in empty plasmid group （ 11.2% ） and PBS group （2. 5% ） （ both P 〈 0. 01 ）. Conclusion EBV-miRNA-BHRF1- 1 can effectively control the mRNA level and protein translation level of p53 gene and may have some inhi- bition effects on nasopharyngeal carcinoma cell proliferation.