Development of novel gene therapy based on the analysis of anti-apoptotic signals in ovarian cancer

在妇科恶性肿瘤中，卵巢癌是最常见的化疗耐药和预后不良的患者。用包括顺铂在内的各种抗癌剂进行化疗的效果显示出由细胞内凋亡信号介导。外源性或内源性抗凋亡因子可能影响化疗效果，并可能导致卵巢癌细胞的化疗耐药。因此，我们的研究目的是分析卵巢癌细胞中的抗凋亡信号，并开发针对这些抗凋亡信号的新的基因治疗方法。我们的研究表明，大约一半的卵巢癌细胞在mRNA和蛋白水平上都表达LH/hCG受体。在表达LH/hCG受体的卵巢癌细胞系OVCAR 3中，hCG通过上调IGF-1抑制顺铂诱导的凋亡。IGF-1还抑制顺铂诱导的细胞凋亡，IGF-1受体的中和抗体恢复了顺铂诱导的细胞凋亡。 ...更多信息 凋亡磷脂酰肌醇3-激酶抑制剂渥曼青霉素阻断IGF-1的抗凋亡作用。因此，促性腺激素是卵巢癌细胞抗凋亡信号的重要外源性因子。用表达反义IGF-1 mRNA的重组腺病毒治疗可有效地恢复顺铂诱导的细胞凋亡。促凋亡基因bax受p53的调控，p53异常作为一种内在的抗凋亡因子导致化疗耐药。在体外，与顺铂敏感的A2780（p53野生型）相比，Bax蛋白在顺铂耐药细胞系如A2780/cDDP（p53突变型）和SKOV 3（p53缺失型）中的表达减弱。我们研制了高表达Bax蛋白的重组腺病毒。Bax/腺病毒转染成功诱导了Bax蛋白的表达，并在卵巢癌顺铂耐药细胞中获得了凋亡效应。因此，腺病毒介导的Bax基因导入可能是一个有用的基因治疗在化疗耐药的卵巢癌。少

Among the gynecological malignant tumors, ovarian carcinoma is most frequently associated with chemoresistance and poor prognosis of the patients. The effect of chemotherapy with various anti-cancer agents including cisplatin is shown to be mediated by intracellular apoptotic signals. Anti-apoptotic factors, either extrinsic or intrinsic, may influence the chemotherapeutic effect, and may result in the chemoresistance of ovarian cancer cells. Therefore, the objective of our study is to analyze the anti-apoptotic signals in ovarian carcinoma cells, and to develop novel gene therapy targeting these anti-apoptotic signals.Our study demonstrated that an approximately half of ovarian carcinomas expressed LH/hCG receptor at both mRNA and protein levels. In ovarian cancer cell line OVCAR3 expressing LH/hCG receptors, hCG inhibited cisplatin-induced apoptosis via up-regulation of IGF-1. IGF-1 also inhibited cisplatin-induced apoptosis and a neutralizing antibody to IGF-1 receptor restored the … More apoptosis. Wortmannin, an inhibitor of phosphatidyl inositol 3-kinase, blocked the anti-apoptotic effect of IGF-1. Therefore, gonadotropin is an important extrinsic factor of anti-apoptotic signal in ovarian cancer cell. Treatment with recombinant adenovirus expressing anti-sense IGF-1 mRNA was effective for restoring the cisplatin-induced apoptosis.Abnormality of p53 tumor suppressor gene is present in more than half of ovarian carcinomas. Pro-apoptotic bax gene is under the regulation of p53, and p53 abnormality resulted in chemoresistance as an intrinsic anti-apototic factor. In vitro, Bax protein expression was attenuated in cisplatin-resistant cell lines such as A2780/cDDP (p53 mutated) and SKOV3 (p53 deleted), compared with cisplatin-sensitive A2780 (p53 wild type). We developed recombinant adenovirus which highly expresses the Bax protein. The Bax protein was successfully induced by treatment with the Bax/adenovirus transfer, and apoptotic effect was obtained in the cisplatin-resistant ovarian cancer cells. Therefore, adenovirus-mediated introduction of Bax may be a useful gene thrapy in the treatment of chemoresistant ovarian carcinomas. Less