人乳头状瘤病毒16型E6E7基因与MCA、TPA协同诱导细胞恶性转化的实验研究

摘要

目的 探讨人乳头状瘤病毒(HPV)16型F6 E7基因与化学致癌物MCA、TPA对Balb/c 3T3细胞恶性转化的协同作用.方法 构建含HPV16 E6E7基因的重组质粒,用其转染Balb/c 3T3细胞.采用RT-PCR和Westem blot技术检测HPV16 E6E7基因和蛋白表达；应用细胞转化实验研究由MCA和TPA诱导的细胞恶性转化；并检验转化细胞在软琼脂上形成集落的能力及对SCID小鼠的致瘤能力.结果 转染HPV16E6E7基因的细胞比未转染细胞形成更多的转化灶,转化灶个数增加4～25倍,且实验时间明显缩短；其转化细胞在软琼脂上形成集落的能力及对SCID小鼠的致瘤能力更强.结论 HPV16 E6E7基因与MCA、TPA可协同诱导Balb/c 3T3细胞恶性转化.%Objective To study carcinogen MCA/TPA induced malignant transformation using Balb/c 3T3 cell trans-fected HPV 16 E6E7 gene. Methods A recombinant plasmid containing HPV16 E6E7 gene was transfected into Balb/c 3T3 cell. The malignant transformation of cells exposed to MCA/TPA was assessed by cell transformation assays. The colony formation in soft agar and tumorigenicity in SCID mice were examined. Results Compared with the cells transfected with vector plasmid, much more transformed foci appeared, much shorter time consumed to accomplish phenotypic alterations , more progressive malignant phenotypic acquired and a bigger diameter of tumors formed in SCID mice in Balb/c 3T3 cells transfected HPV 16 E6E7 gene by the induction of MCA/TPA. Conclusion There could be a synergistic transformation response induced by HPV 16 E6E7 gene and carcinogen MCA/TPA in Balb/c 3T3 cells.