小豆蔻明调控自噬抑制卵巢癌SKOV3细胞增殖的研究

Study on the Inhibitory Effect of Cardamonin on Ovarian Cancer SKOV3 Cells by Regulation of Autophagy

目的:卵巢癌为女性常见病,死亡率高,分子靶向治疗药物较少,研发高效低毒的靶向新药意义重大。细胞自噬(autophagy)维持着细胞内稳态和生长,是抗癌药物作用的关键靶部位。本课题旨在明确小豆蔻明(Cardamonin,CAR)调控自噬对卵巢癌SKOV3细胞增殖的影响。方法:体外培养卵巢癌SKOV3细胞,不同药物分组处理,荧光显微镜下观察单黄酰戊二胺(MDC)染色后细胞自噬囊泡,Western Blot法检测细胞自噬相关蛋白LC3的表达,四氮唑蓝(MTT)法观察SKOV3细胞增殖情况,流式细胞术检测SKOV3细胞凋亡的变化。结果:自噬抑制剂3-MA显著性降低SKOV3细胞内MDC染色的荧光颗粒数目、LC3Ⅱ蛋白表达,抑制细胞增殖;而CAR(高、中剂量)、雷帕霉素和AZD8055与3-MA联用后细胞内MDC荧光颗粒数目增多、LC3Ⅱ蛋白表达增加、细胞增殖抑制率及凋亡率明显升高;且高剂量CAR(10-5mol·L-1)的作用比低剂量CAR(10-6mol·L-1)明显。结论:CAR能够抑制SKOV3细胞增殖,诱导细胞自噬,促进细胞凋亡。CAR有望成为卵巢癌药物治疗的先导化合物。本研究为进一步研发此类化合物提供了实验依据及一定的理论基础。

Objective： Ovarian cancer is a common gynecologic disease with high mortality. Currently, there are few molecular targeted drugs. So it＇s significant to develop efficient new drugs with low toxicity targeted ovarian cancer. The aim of this study is to investigate the effect of Cardamonin （CAR） on ovarian cancer SKOV3 cell proliferation by regulating autophagy. Methods： SKOV3 cells were cultured in vitro and treated with different drugs. The autophagic vesicle with MDC stains were observed by a fluorescence microscope and the protein expression of autophagic protein LC3 was detected by Western blot. Cells proliferation was assayed with MTT method and flow cytometry was used to detect cells apoptosis. Results： The autophagy inhibitor 3-MA significantly decreased the MDC stained fluorescent dots and the expression of LC3 II protein in SKOV3 cells. Cell proliferation was also significantly inhibited by 3-MA. However, both MDC stained fluorescent dots and the expression of LC3 II protein were markedly increased in SKOV3 cells treated by CAR （high- and low-dose）, rapamycin and AZD8055 combined with 3-MA, and, furthermore, cell proliferation inhibition and cell apoptosis were distinctly increased. High-dose of CAR had a stronger effect compared with low-dose. Conclusion： CAR could inhibit the SKOV3 cell proliferation and induce autophagy and apoptosis in SKOV3 cells. CAR is expected to become the lead compound for drug therapy of ovarian cancer. This study provided experimental and theoretical basis for further development of these compounds.