MCT1抑制剂AZD3965增强肝癌细胞对表柔比星敏感性的作用

Monocarboxylic acid transport 1 inhibitor AZD3965 enhances chemosensitivity of hepatocellular carcinoma cells to epirubicin

目的探讨单羧酸转运蛋白1(MCT1)抑制剂AZD3965增强肝癌细胞对表柔比星敏感性的作用。方法 MTT法检测单独使用表柔比星(10,20,40μmol/L)以及20μmol/L AZD3965联合表柔比星(10,20,40μmol/L)处理Hep G2细胞24 h对细胞增殖的影响。使用4μmol/L AZD3965、0. 01μmol/L表柔比星以及4μmol/L AZD3965联合0. 01μmol/L表柔比星处理Hep G2细胞5 d,观察药物对细胞集落克隆形成的影响。用20μmol/L表柔比星处理细胞4 h或20μmol/L表柔比星预处理细胞2 h再与20μmol/L AZD3965联合处理细胞2 h,用活细胞工作站检测细胞内表柔比星的荧光强度;用流式细胞术检测细胞内表柔比星的荧光曲线。结果单独使用表柔比星(10,20,40μmol/L)处理Hep G2细胞24 h的存活率分别为(61. 93±3. 02)%,(59. 87±1. 14)%,(54. 15±6. 14)%。20μmol/L AZD3965作用于Hep G2细胞24 h的存活率为(73. 25±6. 21)%。AZD3965联合表柔比星(10,20,40μmol/L)处理Hep G2细胞24 h,细胞存活率分别为(43. 11±1. 65)%,(45. 27±1. 09)%,(42. 93±1. 86)%。与单独使用表柔比星相比,表柔比星和AZD3965合用后细胞存活率明显降低(P <0. 05)。AZD3965增强表柔比星对Hep G2细胞集落克隆形成的抑制作用。AZD3965增加表柔比星在Hep G2细胞内的分布。AZD3965增加表柔比星在Hep G2细胞内的积累。结论 AZD3965可增强肝癌Hep G2细胞对表柔比星的敏感性,其机制可能是AZD3965增加表柔比星在肝癌细胞内的分布和积累。

Objective To investigate whether AZD3965(monocarboxylic acid transport 1 inhibitor)can enhance the chemosensitivity of hepatocellular carcinoma cells to epirubicin.Methods HepG2 cells were incubated with epirubicin(10,20,40μmol/L),or 20μmol/L AZD3965 combined with Epirubicin(10,20,40μmol/L)for 24 h.Cell viability was measured using MTT assay.Cells were treated with AZD3965(4μmol/L),epirubicin(0.01μmol/L),or AZD3965(4μmol/L)combined with epirubicin(0.01μmol/L)for 5 d,and then the colony formation was detected by crystal violet staining method.Cells were treated with 20μmol/L epirubicin for 4 h,or pretreated with 20μmol/L epirubicin for 2 h and then treated by its combination with 20μmol/L AZD3965 for 2 h.The fluorescence intensity of epirubicin in cell was observed by live cell imaging system.The fluorescence curve of epirubicin in HepG2 cells was examined by flow cytometry.Results The survival rates of HepG2 cells were(61.93±3.02)%,(59.87±1.14)%and(54.15±6.14)%after exposed to 10,20,40μmol/L epirubicin for 24 h,respectively.The survival rate of HepG2 cells was(73.25±6.21)%after exposed to AZD3965(20μmol/L)for 24 h.The survival rates of HepG2 cells were(43.11±1.65)%,(45.27±1.09)%and(42.93±1.86)%after exposed to AZD3965(20μmol/L)combined with 10,20,40μmol/L epirubicin for 24 h,respectively.Compared with epirubicin group,the survival rate of HepG2 cells in combination of epirubicin and AZD3965 group was significantly decreased(P<0.05).AZD3965 enhanced the inhibitory effect of epirubicin on colony formation of HepG2 cells.AZD3965 increased the distribution of epirubicin in HepG2 cells.AZD3965 increased the accumulation of epirubicin in HepG2 cells. Conclusion AZD3965 can enhance the chemosensitivity of HepG2 cells to epirubicin by increasing the distribution and accumulation of epirubicin in HepG2 cells.