Application of antisense oligo DNA to uterine cancers.

子宫颈癌是最常见的生殖器恶性肿瘤，并与人乳头瘤病毒（HPV）的强烈关联已被提出。只有编码E6和E7蛋白的HPV基因组的一部分在癌细胞中始终保留和表达。E6蛋白的一个重要功能是其与细胞p53蛋白形成复合物的能力，导致该字母的泛素依赖性降解。同样，E7癌蛋白也与细胞编码的Rb蛋白形成复合物。在本研究中，首先，我们试图证实宫颈癌与HPV感染的关联。47例原发性宫颈癌中43例（91.5%）存在HPV DNA序列。其中一种癌症含有p53基因突变。此外，其余四种HPV阴性癌症中的一种也含有p53突变。结果，由E6或p53基因突变引起的p53失活对应于44例原发性宫颈癌（93.6%）的发生。我们得到了两个 ...更多信息 原发和复发肿瘤4例。在这些病例中的两个中，观察到在原发性肿瘤中以附加体状态存在的HPV基因组在复发性肿瘤中消失。其中1例复发性肿瘤还存在p53基因突变，提示p53失活可能是HPV感染或p53基因突变所导致的肿瘤侵袭行为的维持。HPV E6和E7的重要性已被确定为包括宫颈癌并维持其表型。因此，我们试图获得HPV 16 E6/E7区反义寡聚体对转化的宫颈上皮细胞系的抑制作用的证据。这些寡聚体大大减少了PHK 16和C4 II细胞的细胞数量和H^3-胸苷摄取。这种有效的抗增殖活性伴随着靶基因表达的抑制。与此相反，SiHa细胞的影响不太明显。这些抑制作用并不伴随着对E7蛋白合成的任何明显抑制。因此，我们通过将作为强DNA加合物的psolaren结合到寡聚体来设计更有效的反义分子。psolaren偶联的寡聚体对SiHa细胞显示出强的抑制作用。与20 μ M硫代磷酸寡聚物相比，psolaren缀合能够将浓度降低至1/40，这显示出相似程度的生长抑制。然而，psolaren缀合的低聚物对正常人表皮角质形成细胞的生长没有显示出任何抑制作用。少

Uterine cervical cancer is the most common genital malignancy, and a strong association with human papillomaviruses (HPV) has been suggested. Only part of the HPV genome, encoding the E6 and E7 proteins, is consistently retained and expressed in Cancer cells. One important function of the E6 protein is its ability to form a complex with the cellular p53 protein, resulting in an ubiguitin-dependent degradation of the letter. Likewise, the E7 oncoprotein also forms a complex with the cell-encoded Rb protein. In the present study, first of all, we tried to confirm the association of cervical cancer with HPV infection. HPV DNA sequences were present in 43 out of 47 primary cervical cancers (91.5%). One of these cancers contained a p53 gene mutation. In addition, One of the remaining four HPV-negative cancers also contained a p53 mutation. As a result, p53 inactivation either by E6 or p53 gene mutations corresponded to the development of 44 primary cervical cancers (93.6%). We obtained both … More primary and recurrent tumors from 4 cases. In two of these cases, the HPV genomes that were present in an episomal state in the primary tumors were observed to have disappeared in the recurrent tumors. One of these recurrent tumors also contained a p53 gene mutation, which suggested the possibility that p53 inactivation was required in order to maintain the aggressive behavior this cancer either by an HPV infection or by a p53 gene umtation.Antisense oligodeoynucleotides (oligomers) have a critical role for dregs targeted specifically against oncogene. The importance of HPV E6 and E7 has been identified to include the cervical cancer and maintain its phenotype. Thus, we tried to obtain evidence for the inhibitory effects of antisense oligomers to the E6/E7 region of HPV16 on transformed cervical epithelial cell lines. The oligomers greatly reduced both cell numbers and H^3-Thymidine uptake in PHK16 and C4II cells. This potent antiproliferative activity accompanied the suppression of target gene expression. In contrast, the effects on SiHa cells were less remarkable. These inhibitory effects were not accompanied any noticeable inhibition of E7 protein synthesis. Thus, we design more efficient antisense molecules by binding the psolaren that is a strong DNA adduct, to the oligomors. The psolaren-conjugated oligomers showed the strong inhibitory effects on SiHa cells. The psolaren conjugation was able to lessen the concentration into 1/40, that showed the similar degree of growth inhibition, compared to the 20muM phospborothioate oligomers. However, the psolaren conjugated oligomers did not show any inhibitory effect on growth of the normal human epidermal karatinocytes. Less