黄芩素对顺铂抑制卵巢癌A2780细胞增殖的影响  

Effect of baicalein on the inhibitory role of cisplatin on the proliferation of ovarian cancer A2780 cells

作　　者：郑宁泽 陶亮 王琴 Zheng Ningze;Tao Liang;Wang Qin(Department of Pharmacology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]中山大学中山医学院药理学教研室,广州510080

出　　处：《新医学》2020年第5期348-353,共6页Journal of New Medicine

基　　金：广东省自然科学基金(2019A1515012215)。

摘　　要：目的体外检测黄芩素对顺铂抑制卵巢癌A2780细胞增殖的影响及其机制。方法选择对数生长期A2780细胞,采用CCK-8法分别检测黄芩素对A2780细胞的毒性、有黄芩素和无黄芩素作用下顺铂对A2780细胞的半数抑制浓度(IC50)差异、在细胞缝隙连接(GJ)通道功能抑制剂18-α-甘草次酸(18-α-GA)的作用下顺铂对A2780细胞抑制率的变化以及增加黄芩素对顺铂作用的影响,使用细胞接种荧光示踪法检测黄芩素对A2780细胞GJ通道功能的影响。利用GEO公共数据库,比较卵巢正常上皮组织和卵巢癌上皮组织中编码Cx43的GJA1基因表达的差异。利用蛋白免疫印迹法检测黄芩素对A2780细胞中Cx43蛋白表达的影响。利用Swiss Target Prediction数据库预测黄芩素的靶蛋白,并构建靶蛋白与GJ通道的互作网络。结果当黄芩素≤10.00μmol/L,其对A2780细胞无明显毒性。与单用顺铂比较,10.00μmol/L黄芩素可增加顺铂对A2780细胞的抑制率(P <0.01)。GEO公共数据库分析显示,卵巢癌上皮组织中GJA1 mRNA的表达水平低于卵巢正常上皮组织(P <0.01);黄芩素可上调A2780细胞中Cx43蛋白表达量(P <0.01)。18-α-GA可降低顺铂对A2780细胞的抑制率,并且能逆转黄芩素对顺铂增敏作用(P均<0.01)。Swiss Target Prediction数据库分析显示,黄芩素直接靶向于GJ通道中的4个蛋白。结论黄芩素能增强顺铂对卵巢癌A2780细胞的毒性,其机制可能与增强A2780细胞的GJ通道功能有关。Objective To evaluate the effect and mechanism of baicalein on the inhibitory role of cicplatin on the proliferation of ovarian cancer A2780 cells. Methods The A2780 cells in the logarithmic growth phase were selected. The cytotoxicity of baicalein on the ovarian cancer A2780 cells was assessed by CCK-8 assay. The difference of IC50 of cisplatin in the A2780 cells treated with and without baicalein was statistically compared. After the treatment with cell gap junction(GJ) pathway inhibitor 18-α-glycyrrhetinic acid(18-α-GA), the effect of cisplatin on the inhibitory rate of A2780 cells was evaluated. The effect of baicalein treatment on the inhibitory role of cisplatin was assessed. The impact of baicalein on the function of GJ signaling pathway in the A2780 cells was evaluated by Parachute assay. Based on the GEO dataset, the expression levels of GJA1(Cx43) genes between the normal ovarian epithelial and ovarian cancer epithelial tissues were statistically compared. The effect of baicalein on the expression of Cx43 protein in the A2780 cells was assessed by Western blot. The targeted protein of baicalein was predicted by Swiss Target Prediction database. The interaction network between the targeted proteins and GJ pathway was constructed. Results Baicalein treatment at a dose of≤10 μmol/L provoked no obvious toxicity to the A2780 cells. Compared with cisplatin alone, baicalein treatment at a dose of 10 μmol/L could significantly increase the inhibitory rate of cisplatin on the A2780 cells(P < 0.01). Administration of 18-α-GA could considerably lower the inhibitory rate of cisplatin on the A2780 cells and significantly reverse the sensitization effect of baicalein on cisplatin(both P < 0.01). GEO dataset revealed that the expression level of Cx43 m RNA in the ovarian cancer epithelial tissues was significantly lower compared with that in the normal ovarian epithelial tissues(P < 0.05). Treatment with baicalein could significantly up-regulate the expression level of Cx43 protein(P < 0.01). Swiss Target Predi

关 键 词：黄芩素 卵巢癌 顺铂 细胞缝隙连接 

分 类 号：R737.31[医药卫生—肿瘤]