Suppression of tumorigenesis by transcription units expressing the antisense E6 and E7 messenger RNA (mRNA) for the transforming proteins of the human papilloma virus and the sense mRNA for the retinoblastoma gene in cervical carcinoma cells.
Suppression of tumorigenesis by transcription units expressing the antisense E6 and E7 messenger RNA (mRNA) for the transforming proteins of the human papilloma virus and the sense mRNA for the retinoblastoma gene in cervical carcinoma cells.
复制标题
通过表达人乳头瘤病毒转化蛋白的反义 E6 和 E7 信使 RNA (mRNA) 以及宫颈癌细胞中视网膜母细胞瘤基因的有义 mRNA 的转录单位抑制肿瘤发生。
作者：
G. Hu;W. Liu;E. Hanania;Siqing Fu;T. Wang;A. Deisseroth
据报道，携带人乳头状瘤病毒（HPV）的人宫颈癌细胞系至少在疾病的开始阶段（如果不是在所有阶段）表达HPV E6和E7蛋白。HPV E6和E7蛋白与p53和视网膜母细胞瘤（Rb）肿瘤抑制基因的产物结合并结合，从而使宫颈癌细胞避开这些肿瘤抑制基因的作用。我们观察到，将反义HPV 18 E6和E7序列以及人野生型Rb基因的正义cDNA导入人宫颈癌细胞系（HeLa），该细胞系对HPV 18前病毒呈阳性，如果HPV 18 E6和E7的反义转录物和人Rb的有义转录物都是表达。此外，Rb信使RNA的互补DNA（cDNA）的过表达足以减缓HeLa细胞的增殖，并且Rb cDNA的表达水平与肿瘤生长速率减缓的程度相关。我们的实验结果表明，HPV E6和E7蛋白的存在和由此产生的Rb在宫颈癌细胞中的失活有助于肿瘤表型，即使在高度进化的宫颈癌细胞系，如HeLa，其已来自宫颈癌患者在疾病过程的晚期。这些数据表明，HPV蛋白不仅在宫颈癌的开始阶段发挥作用，而且在这种疾病的晚期阶段也发挥作用。这些实验可能会导致遗传方法来控制这种疾病，涉及反义序列下调E6和E7基因或导致Rb基因的表达。
Human cervical carcinoma cell lines that harbor human papilloma virus (HPV) have been reported to express HPV E6 and E7 proteins at least in the beginning stages if not at all stages of the disease. The HPV E6 and E7 proteins bind to and inactivate the products of the p53 and retinoblastoma (Rb) tumor suppressor genes, which thereby allow the cervical carcinoma cells to circumvent the action of these tumor suppressor genes. We observed that the introduction of the antisense HPV 18 E6 and E7 sequences, as well as a sense cDNA for the human wild-type Rb gene into a human cervical carcinoma cell line (HeLa), which is positive for the HPV 18 provirus, decreased the in vitro and in vivo growth rate of the transfected cells if both antisense transcripts for the HPV 18 E6 and E7 and sense transcripts for human Rb were expressed. In addition, overexpression of a complementary DNA (cDNA) for the Rb messenger RNA was sufficient to slow the proliferation of HeLa cells, and the level of Rb cDNA expression was correlated with the degree to which the rate of growth of the tumor was slowed. The results of our experiments show that the presence of HPV E6 and E7 proteins and the resultant inactivation of Rb in cervical carcinoma cells contributes to the neoplastic phenotype even in highly evolved cervical carcinoma cell lines such as HeLa, which have been derived from a cervical carcinoma patient at an advanced stage of the disease process. These data suggest that the HPV proteins play a role not only at the beginning of cervical cancer, but also at advanced stages of this disease. These experiments may lead to genetic approaches to the control of this disease that involve antisense sequences that downregulate the E6 and E7 genes or lead to expression of the Rb gene.