Shen Wei, Liang Bingfeng, Yin Jie, Li Xiurong, Cheng Jianxin
Department of Obstetrics and Gynecology, The Fourth Hospital of Hebei Medical University, 12 Jiankang Road, Shijiazhuang, 050011, Hebei, China.
Department of Nursing, Hebei Women Vocational Technology College, 16 Huiwen Street, Shijiazhuang, 050091, Hebei, China.
Cell Biochem Biophys. 2015 May;72(1):203-13. doi: 10.1007/s12013-014-0438-y.
Cisplatin is a first-line chemotherapy drug against ovarian cancer. However, its strong toxic side effects and the development of cisplatin resistance in human cancer cells seriously influence the effects of chemotherapy and quality of life in patients. Noscapine (Nos), a non-toxic benzylisoquinoline alkaloid extracted from opium, has been recently reported to have anti-cancer activity, but the mechanism of that effect has not been clearly established. In the present study, we investigated cytotoxicity of Nos in combination with cisplatin (DDP) in drug-resistant human ovarian cancer cell line SKOV3/DDP in vitro and in vivo null mice xenograft model. Cell proliferation was measured by MTT assay, flow cytometry was used to analyze cell cycle and apoptosis, protein expression of several apoptotic factors was investigated by flow cytometry and immunohistochemical method, and their mRNA expression levels were determined by real-time PCR. In vitro experiments showed that Nos significantly inhibited proliferation of SKOV3/DDP cells. DDP/Nos-combined treatment notably enhanced DDP-induced inhibition of cell proliferation and increased the pro-apoptotic effect of DDP in SKOV3/DDP cells. DDP/Nos administration increased the proportion of G2/M cells, reduced both protein and mRNA expression of anti-apoptotic factors XIAP, surviving and NF-kB, and augmented protein and mRNA levels of pro-apoptotic caspase-3. In vivo experiments revealed that Nos/DDP treatment increased the apoptotic rate of xenograft tumors in null mice. Tumor volume decreased from 1.733 ± 0.155 g in mice treated with DDP alone to 1.191 ± 0.106 g in animals treated with Nos/DDP. These observations suggest that Nos increases the anti-cancer activity of DDP against the drug-resistant ovarian cancer cell line SKOV3/DDP by modulating the cell cycle and activating apoptotic pathways. The study provides a new chemotherapy strategy for the treatment of DDP-resistant human ovarian cancer.
顺铂是治疗卵巢癌的一线化疗药物。然而，其强烈的毒副作用以及人类癌细胞中顺铂耐药性的产生，严重影响了化疗效果和患者的生活质量。那可丁（Nos）是一种从鸦片中提取的无毒苄基异喹啉生物碱，最近有报道称其具有抗癌活性，但其作用机制尚未明确。在本研究中，我们在体外对耐药人卵巢癌细胞系SKOV3/DDP以及体内裸鼠异种移植模型中，研究了那可丁与顺铂（DDP）联合使用时的细胞毒性。通过MTT法检测细胞增殖，采用流式细胞术分析细胞周期和凋亡情况，通过流式细胞术和免疫组化方法研究几种凋亡因子的蛋白表达，并通过实时PCR测定其mRNA表达水平。体外实验表明，那可丁显著抑制SKOV3/DDP细胞的增殖。DDP/Nos联合治疗显著增强了DDP诱导的细胞增殖抑制作用，并增加了DDP对SKOV3/DDP细胞的促凋亡作用。DDP/Nos给药增加了G2/M期细胞的比例，降低了抗凋亡因子XIAP、存活蛋白和NF-κB的蛋白及mRNA表达，并增加了促凋亡半胱天冬酶-3的蛋白和mRNA水平。体内实验显示，Nos/DDP治疗增加了裸鼠异种移植肿瘤的凋亡率。肿瘤体积从单独使用DDP治疗的小鼠中的1.733±0.155 g降至Nos/DDP治疗的动物中的1.191±0.106 g。这些观察结果表明，那可丁通过调节细胞周期和激活凋亡途径，增强了DDP对耐药卵巢癌细胞系SKOV3/DDP的抗癌活性。该研究为治疗顺铂耐药的人类卵巢癌提供了一种新的化疗策略。
