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目的 研究EB病毒LMP1反义基因对人鼻咽癌CNE2细胞恶性表型的逆转作用。方法 用反义LMP1DNA的真核表达载体anti pcDNA3.1 LMP1转染人鼻咽癌细胞CNE2 ,G4 18筛选抗性克隆 ;Western blot法检测转染前、后瘤细胞的LMP1表达 ;用MTT法、细胞生长曲线和软琼脂集落形成能力实验观察转染前后细胞体外生长增殖特性。结果 建立反义CNE2细胞克隆 ;转染后LMP1蛋白表达降低 ;转染后细胞活力和生长速度均有下降 ,集落形成能力显著降低。结论 将反义LMP1真核表达载体导入细胞能成功抑制NPC细胞的生长 ,逆转其恶性表型。本研究还为鼻咽癌的基因治疗提供了实验依据。
Objective To study the reversal effect of EBV LMP1 antisense gene on the malignant phenotype of human nasopharyngeal carcinoma CNE2 cells. Methods Antisense LMP1 DNA was transfected into human nasopharyngeal carcinoma cell lines CNE2 and G4 18 using anti-pcDNA3.1 LMP1. The expression of LMP1 was detected by Western blot. The expression of LMP1 was detected by MTT assay. Cell growth curve and soft agar colony formation ability were observed before and after transfection cells growth and proliferation characteristics. Results The antisense CNE2 cell clone was established. The expression of LMP1 protein was decreased after transfection. The viability and growth rate of transfected cells decreased and the colony formation ability was significantly decreased. Conclusion The antisense LMP1 eukaryotic expression vector can successfully inhibit the growth of NPC cells and reverse its malignant phenotype. This study also provides experimental evidence for gene therapy of NPC.