Giri Shailendra, Rattan Ramandeep, Deshpande Mandar, Maguire Jacie L, Johnson Zachary, Graham Rondell P, Shridhar Viji
Department of Experimental Pathology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, Minnesota, United States of America.
Women's Health Services, Henry Ford Health System, Detroit, Michigan, United States of America.
PLoS One. 2014 Jun 2;9(6):e97897. doi: 10.1371/journal.pone.0097897. eCollection 2014.
This study examines the role of s-nitrosylation in the growth of ovarian cancer using cell culture based and in vivo approaches. Using the nitrosylating agent, S-nitrosoglutathione (GSNO), a physiological nitric oxide molecule, we show that GSNO treatment inhibited proliferation of chemoresponsive and chemoresistant ovarian cancer cell lines (A2780, C200, SKVO3, ID8, OVCAR3, OVCAR4, OVCAR5, OVCAR7, OVCAR8, OVCAR10, PE01 and PE04) in a dose dependent manner. GSNO treatment abrogated growth factor (HB-EGF) induced signal transduction including phosphorylation of Akt, p42/44 and STAT3, which are known to play critical roles in ovarian cancer growth and progression. To examine the therapeutic potential of GSNO in vivo, nude mice bearing intra-peritoneal xenografts of human A2780 ovarian carcinoma cell line (2 × 10(6)) were orally administered GSNO at the dose of 1 mg/kg body weight. Daily oral administration of GSNO significantly attenuated tumor mass (p<0.001) in the peritoneal cavity compared to vehicle (phosphate buffered saline) treated group at 4 weeks. GSNO also potentiated cisplatin mediated tumor toxicity in an A2780 ovarian carcinoma nude mouse model. GSNO's nitrosylating ability was reflected in the induced nitrosylation of various known proteins including NFκB p65, Akt and EGFR. As a novel finding, we observed that GSNO also induced nitrosylation with inverse relationship at tyrosine 705 phosphorylation of STAT3, an established player in chemoresistance and cell proliferation in ovarian cancer and in cancer in general. Overall, our study underlines the significance of S-nitrosylation of key cancer promoting proteins in modulating ovarian cancer and proposes the therapeutic potential of nitrosylating agents (like GSNO) for the treatment of ovarian cancer alone or in combination with chemotherapeutic drugs.
本研究采用基于细胞培养和体内实验的方法,探讨了S-亚硝基化在卵巢癌生长中的作用。使用亚硝基化剂S-亚硝基谷胱甘肽(GSNO),一种生理性一氧化氮分子,我们发现GSNO处理以剂量依赖性方式抑制了化疗敏感和化疗耐药的卵巢癌细胞系(A2780、C200、SKVO3、ID8、OVCAR3、OVCAR4、OVCAR5、OVCAR7、OVCAR8、OVCAR10、PE01和PE04)的增殖。GSNO处理消除了生长因子(HB-EGF)诱导的信号转导,包括Akt、p42/44和STAT3的磷酸化,已知这些在卵巢癌的生长和进展中起关键作用。为了研究GSNO在体内的治疗潜力,将携带人A2780卵巢癌细胞系(2×10⁶)腹腔异种移植瘤的裸鼠,以1mg/kg体重的剂量口服给予GSNO。与载体(磷酸盐缓冲盐水)处理组相比,在4周时,每日口服GSNO显著减轻了腹腔内的肿瘤质量(p<0.001)。GSNO还增强了顺铂在A2780卵巢癌裸鼠模型中介导的肿瘤毒性。GSNO的亚硝基化能力反映在包括NFκB p65、Akt和EGFR在内的各种已知蛋白质的诱导亚硝基化上。作为一项新发现,我们观察到GSNO还诱导了STAT3酪氨酸705磷酸化的亚硝基化,且二者呈负相关,STAT3是卵巢癌及一般癌症中化疗耐药和细胞增殖的既定参与者。总体而言,我们的研究强调了关键癌症促进蛋白的S-亚硝基化在调节卵巢癌中的重要性,并提出了亚硝基化剂(如GSNO)单独或与化疗药物联合治疗卵巢癌的治疗潜力。