EBV—miRNA—BHRF1—1对鼻咽癌细胞p53基因表达的影响

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能有效的调控声3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。

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

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.