多种含硒复合物对犬乳腺癌细胞CTM1211的抑制作用及其机制

Inhibition and related mechanisms of different selenium compounds on canine breast cancer cells CTM1211

研究不同剂量、形式、配伍的含硒复合物对犬乳腺癌细胞CTM1211的影响,以探究硒抗癌的有效剂量、形式和机制。用低、中、高剂量的CTX(环磷酰胺,1、2、4mg/mL)、SSE(亚硒酸钠,10、20、40μmol/L)、MSA(甲亚硒酸,10、20、40μmol/L)、MSC(硒代半胱氨酸,200、400、800μmol/L)、CTX+SSE(0.5 mg/mL+5μmol/L、1mg/mL+10μmol/L、2mg/mL+20μmol/L)、CTX+MSA(0.5mg/mL+5μmol/L、1mg/mL+10μmol/L、2mg/mL+20μmol/L)、CTX+MSC(0.5mg/mL+100μmol/L、1mg/mL+200μmol/L、2mg/mL+400μmol/L)分别作用CTM1211细胞24、48、72h,MTT法检测以上各组细胞存活率;用CTX(2 mg/mL)、SSE(40μmol/L)、MSA(20μmol/L)、MSC(400μmol/L)、CTX+MSA(2 mg/mL+20μmol/L)分别作用CTM1211细胞48h,流式细胞仪检测各组细胞凋亡率,免疫组化法及RT-qPCR法分别检测VEGF-a(血管内皮生长因子a)、PTEN(磷酸酯酶与张力蛋白同源物)、Ang-2(血管生成素2)、HIF-1a(缺氧诱导因子1a)蛋白和mRNA的表达。与对照组比较,各硒作用组的细胞存活率在48h/72h时显著降低(P<0.01或P<0.05)、凋亡率显著增高(P<0.01),其中CTX+MSA组效果最显著;VEGF-a、Ang-2和HIF-1a蛋白及mRNA的表达在整体上被显著下调,PTEN蛋白及mRNA的表达在整体上被显著上调。结果表明,硒尤其是MSA能显著抑制犬乳腺癌细胞CTM1211,其作用机制与诱导凋亡和调节肿瘤血管生长相关因子VEGF-a、PTEN、Ang-2、HIF-1a有关。

To evaluate the effect of different doses,forms and compatibility of selenium on canine breast cancer cells CTM1211 and to explore the related mechanisms,CTM1211 cells were intervened by different doses of CTX(cyclophosphamide:1,2,4 mg/mL),SSE(selenite:10,20,40μmol/L),MSA(methylseleninic acid:10,20,40μmol/L),MSC(selenocysteine:200,400,800μmol/L),CTX+SSE(0.5mg/mL+5μmol/L,1 mg/mL+10μmol/L,2 mg/mL+20μmol/L),CTX+ MSA(0.5 mg/mL+5μmol/L,1mg/mL+10μmol/L,2mg/mL+20μmol/L)and CTX+MSC(0.5mg/mL+100μmol/L,1mg/mL+200μmol/L,2mg/mL+400μmol/L)for 24 h,48hand 72 h,and the cell viability of each group was determined by MTT.While simplified cells were intervened by CTX(2 mg/mL),SSE(40μmol/L),MSA(20μmol/L),MSC(400μmol/L)and CTX+MSA(2mg/mL+20μmol/L)for 48 hand the cell apoptosis rate was measured by flow cytometry,the protein and mRNA expression of VEGF-a(vascular endothelial growth factor a),PTEN(phosphatase and tensin homolog),Ang-2(angiopoietin-2)and HIF-1a(hypoxia inducible factor 1a)were measured by immunohistochemistry and RT-qPCR,respectively.The cell viability of each group at 48h/72 hwas significantly lower(P<0.01 or P<0.05),while the apoptosis ratewas significantly higher at each time point(P<0.01)than that of the control group,and the effect of group CTX + MSA was the most significant.Generally,the expression of VEGF-a,Ang-2and HIF-1aprotein and mRNA were significantly low,while that of PTEN were significantly up regulated.In conclusion,selenium,especially MSA,could significantly inhibit breast cancer cell CTM1211,which is partly due to the induction of apoptosis and regulation of tumor angiogenesis-related factors VEGF-a,PTEN,Ang-2and HIF-1aby selenium.