姜黄素对人食管癌耐药细胞逆转作用的实验研究  

Curcumin Reversed Human Esophageal Carcinoma Drug Resistant Cells

作　　者：刘华[1] 白美玲[2] 郭颖[2] 张晓丽[1] 张林西[1] 罗强[1] 孙黎[1] 

机构地区：[1]河北北方学院生命科学研究中心,河北075000 [2]河北北方学院基础医学院病理学教研室,河北075000

出　　处：《中国中西医结合杂志》2017年第5期557-562,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：河北省自然科学基金资助项目(No.H2014405033);河北北方学院重大课题资助项目(No.ZD201414)

摘　　要：目的研究姜黄素对食管癌耐药细胞的作用及作为食管癌多药耐药逆转剂的可能性。方法采用20、40、80μmol/L姜黄素作用体外培养Eca-109/Taxol细胞及Eca-109细胞,倒置显微镜观察姜黄素对Eca-109/Taxol细胞作用的形态变化,CCK-8法检测姜黄素对两种细胞的增殖抑制率并计算姜黄素的逆转效应。流式细胞术检测姜黄素对Eca-109/Taxol细胞的凋亡率及测定罗丹明123在癌细胞内的蓄积。采用全自动酶标仪检测Caspase-3活性、免疫细胞化学法检测P-gp在癌细胞中的表达。结果 Eca-109/Taxol细胞经不同浓度姜黄素作用后,细胞生长变慢、细胞变圆,部分细胞悬浮于培养液中。随着姜黄素浓度的增高,对Eca-109/Taxol细胞生长抑制作用逐渐增强。姜黄素对Eca-109/Taxol细胞具有明显的耐药逆转作用,10μmol/L姜黄素逆转2.50倍,40μmol/L姜黄素逆转6.83倍,且在一定浓度范围内呈剂量依赖性。姜黄素与低浓度Taxol联合应用,可使Eca-109/Taxol细胞中Caspase-3活力提高。流式细胞仪检测显示,随姜黄素浓度的增加,癌细胞的凋亡率逐步增高,20μmol/L姜黄素作用24 h后凋亡率为(13.6±1.3)%,80μmol/L姜黄素凋亡率为(43.5±1.6)%,组间比较,差异有统计学意义(P<0.01),同时可使Eca-109/Taxol细胞内Rho123的蓄积增高(P<0.01);20μmol/L姜黄素对Eca-109/Taxol细胞作用24 h可明显抑制P-gp在Eca-109/Taxol细胞中的表达。结论姜黄素对Eca-109/Taxol细胞生长具有抑制作用,可以较强地逆转癌细胞对Taxol耐药性,可能是食管癌临床化疗中较好的耐药逆转药物。Objective To study the effects of curcumin on esophageal carcinoma celldrugresis- tant cells, and to study its possibility as reversal agent. Methods The cell lines of Eca-109/Taxol and Eca-109 were cultured by 20, 40, and 80 μmol/L curcumin in vitro. Morphological changes of Eca-109/ Taxol cells treated by curcumin were observed under inverted microscope. The proliferative inhibition rates of curcumin against two cell lines were detected by CCK-8 assay and the reversal effect of curcu- min was calculated. The apoptosis rate of Eca-109/Taxol cells was detectedby flow cytometry. The accu- mulation of Rho123 in cancer cells was determined. Caspase-3 activity was detected by automatic micro- plate reader. The expression of P-gp in cancer cells was detected by immunocytochemistry. Results Eca-109/Taxol cells grew slowly and became round. Partial cells were suspended in the culture solution after treated by different concentrations of curcumin. The inhibition effect on Eca-109/Taxol cell growth was gradually enhanced as increased concentrations of curcumin. Curcumin had significantly reversal effect on Eca-109/Taxol cells, 2.50 times by 10 i^mol/L curcumin and 6.83 times by 40 mol/L curcumin, showing dose dependent manner in certain concentration ranges. Combining curcumin with low concen- tration Taxol could elevate the activity of Caspase-3 in Eca-109/Taxol cells. Flow cytometry showed that the apoptosis rates gradually increased with rising concentration of curcumin. The apoptosis rates were 13.6%±1.3% and 43.5 ±1.6% respectively after treated by 20 i^mol/L and 80 μmol/L curcumin for 24 h, with statistical difference between the two groups （P 〈0.01 ）. Meanwhile, Rho123 accumulation was in- creased in Eca-109/Taxol cells（P 〈0.01 ）. Expression of P-gp in Eca-109/Taxol cells after treated by 20 μmol/L curcumin for 24 h could be markedly inhibited. Conclusions Curcumin could inhibit the growth of Eca-lO9/Taxol cells, and reverse thier drug resistance to Taxol. It might be a good reversal agent to drug

关 键 词：食管癌 姜黄素 多药耐药 细胞凋亡 

分 类 号：R285[医药卫生—中药学]