姜黄素对乳腺癌细胞顺铂敏感性的影响
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摘要
目的探讨姜黄素(curcumin)对乳腺癌细胞中顺铂(Cisplatin,CDDP)治疗敏感性的影响及其可能机制。方法 CCK-8检测、Hoechst染色观察CDDP、姜黄素单独及联合用药48 h对MCF7细胞增殖抑制和细胞凋亡的影响;Western blot分析MCF7细胞在CDDP(0、1.25、2.5、5、10、20μg/m L)、姜黄素(0、1、5、20、30、50、100μmol/L)单独及联合处理后FEN1(flap endonuclease 1)表达水平的变化;CCK-8检测沉默FEN1对MCF7细胞中CDDP敏感性的影响。结果 CDDP、姜黄素均可以剂量依赖方式抑制MCF7细胞增殖,其IC50分别为5、30μmol/L;与单独2μg/m L CDDP处理组比较,2μg/m L CDDP联合20μmol/L姜黄素、30μmol/L姜黄素处理组对细胞增殖的抑制效应明显增强[分别为(84.1±0.8)%、(51.1±0.5)%、(29.4±0.3)%,P<0.05];与单独20μmol/L姜黄素处理组比较,20μmol/L姜黄素联合2μg/m L CDDP、5μg/m L CDDP处理组对细胞增殖的抑制效应也明显增强[分别为(76.9±0.7)%、(42.4±0.3)%、(31.6±0.4)%,P<0.05];2μg/m L CDDP联合20μmol/L姜黄素组的细胞凋亡明显增强(P<0.05);与Control siRNA组比较,FEN1 siRNA+CDDP组可显著增强CDDP抑制MCF7细胞增殖的敏感性[(25.4±0.3)%vs(18.7±0.2)%,P<0.05];CDDP增殖抑制无效浓度或低浓度时,FEN1蛋白的表达随CDDP的处理剂量依赖性上调,而姜黄素可剂量依赖性下调FEN1蛋白表达;与单独CDDP和姜黄素处理组比较,2μg/m L CDDP联合20μmol/L姜黄素组可显著降低FEN1蛋白表达(P<0.05)。结论姜黄素可增强CDDP对乳腺癌细胞的敏感性,其机制与降低细胞FEN1的表达有关。
Objective To explore the effect of curcumin on the sensitivity of breast cancer cells to cisplatin( CDDP) and possible mechanism. Methods CCK-8 assay and Hoechst staining were used to observe the proliferation and apoptosis of MCF-7 cells treated with curcumin alone or in combination with CDDP for 48 h. Western blotting was used to detect the expression level of flap endonuclease 1( FEN1) in MCF7 cells treated as above. The sensitivity of FEN1-silenced MCF-7 cells to CDDP was detected by CCK-8assay. Results Both CDDP and curcumin could inhibit MCF-7 cell proliferation in a dose-dependent manner,and the IC50 values of CDDP and curcumin were 5 and 30 μmol / L,respectively. Compared with the cells treated with 2 μg / m L CDDP alone,the proliferation of the cells treated with CDDP in combination with20 or 30 μmol / L curcumin was significantly inhibited [( 84. 1 ± 0. 8) % vs( 51. 1 ± 0. 5) %,( 84. 1 ± 0. 8) %vs( 29. 4 ± 0. 3) %,respectively,P < 0. 05]. Compared with the cells treated with curcumin alone,the proliferation of the cells treated with 20 μmol / L curcumin in combination with 2 or 5 μg / m L CDDP was also significantly inhibited [( 76. 9 ± 0. 7) % vs( 542. 4 ± 0. 3) %,( 76. 9 ± 0. 7) % vs( 31. 6 ± 0. 4) %,respectively,P < 0. 05]. Apoptosis of the cells treated with 2 μg / m L CDDP in combination with 20 μmol / L curcumin was obviously increased( P < 0. 05). Compared with control treatment,silencing FEN1 with siRNA could enhance the sensitivity of MCF7 cell to CDDP( P < 0. 05). FEN1 protein was up-regulated in a dosedependent manner in the cells treated with low concentration or invalid concentration of CDDP. However,curcumin could obviously down-regulate FEN1 protein in a dose-dependent manner. Compared with the cells treated with CDDP or curcumin alone,FEN1 protein expression was significantly reduced in the cells treated with 2 μg / m L CDDP in combination with 20 μmol / L curcumin( P < 0. 05). Conclusion Curcumin can enhance the sensitivity of breast cancer cells to CDDP,which is related with the decrease of FEN1 expression.
Objective To explore the effect of curcumin on the sensitivity of breast cancer cells to cisplatin( CDDP) and possible mechanism. Methods CCK-8 assay and Hoechst staining were used to observe the proliferation and apoptosis of MCF-7 cells treated with curcumin alone or in combination with CDDP for 48 h. Western blotting was used to detect the expression level of flap endonuclease 1( FEN1) in MCF7 cells treated as above. The sensitivity of FEN1-silenced MCF-7 cells to CDDP was detected by CCK-8assay. Results Both CDDP and curcumin could inhibit MCF-7 cell proliferation in a dose-dependent manner,and the IC50 values of CDDP and curcumin were 5 and 30 μmol / L,respectively. Compared with the cells treated with 2 μg / m L CDDP alone,the proliferation of the cells treated with CDDP in combination with20 or 30 μmol / L curcumin was significantly inhibited [( 84. 1 ± 0. 8) % vs( 51. 1 ± 0. 5) %,( 84. 1 ± 0. 8) %vs( 29. 4 ± 0. 3) %,respectively,P < 0. 05]. Compared with the cells treated with curcumin alone,the proliferation of the cells treated with 20 μmol / L curcumin in combination with 2 or 5 μg / m L CDDP was also significantly inhibited [( 76. 9 ± 0. 7) % vs( 542. 4 ± 0. 3) %,( 76. 9 ± 0. 7) % vs( 31. 6 ± 0. 4) %,respectively,P < 0. 05]. Apoptosis of the cells treated with 2 μg / m L CDDP in combination with 20 μmol / L curcumin was obviously increased( P < 0. 05). Compared with control treatment,silencing FEN1 with siRNA could enhance the sensitivity of MCF7 cell to CDDP( P < 0. 05). FEN1 protein was up-regulated in a dosedependent manner in the cells treated with low concentration or invalid concentration of CDDP. However,curcumin could obviously down-regulate FEN1 protein in a dose-dependent manner. Compared with the cells treated with CDDP or curcumin alone,FEN1 protein expression was significantly reduced in the cells treated with 2 μg / m L CDDP in combination with 20 μmol / L curcumin( P < 0. 05). Conclusion Curcumin can enhance the sensitivity of breast cancer cells to CDDP,which is related with the decrease of FEN1 expression.
引文
[1]Wilmes A,Bielow C,Ranninger C,et al.Mechanism of cisplatin proximal tubule toxicity revealed by integrating transcriptomics,proteomics,metabolomics and biokinetics[J].Toxicol In Vitro,2015,30(1 Pt A):117-127.DOI:10.1016/j.tiv.2014.10.006
[2]Ugur S,Ulu R,Dogukan A,et al.The renoprotective effect of curcumin in cisplatin-induced nephrotoxicity[J].Ren Fail,2015,37(2):332-336.DOI:10.3109/0886022X.2014.986005
[3]Wang J,Zhou L,Li Z,et al.YY1 suppresses FEN1 over-expression and drug resistance in breast cancer[J].BMC Cancer,2015,15:50.DOI:10.1186/s12885-015-1043-1
[4]Balakrishnan L,Bambara R A.Flap endonuclease 1[J].Annu Rev Biochem,2013,82:119-138.DOI:10.1146/annurev-biochem-072511-122603
[5]Abdel-Fatah T M,Russell R,Albarakati N,et al.Genomic and protein expression analysis reveals flap endonuclease 1(FEN1)as a key biomarker in breast and ovarian cancer[J].Mol Oncol,2014,8(7):1326-1338.DOI:10.1016/j.molonc.2014.04.009
[6]Chereddy K K,Coco R,Memvanga P B,et al.Combined effect of PLGA and curcumin on wound healing activity[J].J Control Release,2013,171(2):208-215.DOI:10.1016/j.jconrel.2013.07.015
[7]Hatcher H,Planalp R,Cho J,et al.Curcumin:from ancient medicine to current clinical trials[J].Cell Mol Life Sci,2008,65(11):1631-1652.DOI:10.1007/s00018-008-7452-4
[8]Chang Z,Xing J,Yu X.Curcumin induces osteosarcoma MG63 cells apoptosis via ROS/Cyto-C/Caspase-3 pathway[J].Tumour Biol,2014,35(1):753-758.DOI:10.1007/s13277-013-1102-7
[9]Lv Z D,Liu X P,Zhao W J,et al.Curcumin induces apoptosis in breast cancer cells and inhibits tumor growth in vitro and in vivo[J].Int J Clin Exp Pathol,2014,7(6):2818-2824.
[10]Chen B,Zhang Y,Wang Y,et al.Curcumin inhibits proliferation of breast cancer cells through Nrf2-mediated downregulation of Fen1 expression[J].J Steroid Biochem Mol Biol,2014,143:11-18.DOI:10.1016/j.jsbmb.2014.01.009
[11]Kelland L.The resurgence of platinum-based cancer chemotherapy[J].Nat Rev Cancer,2007,7(8):573-584.DOI:10.1038/nrc2167
[12]Armstrong D K,Bundy B,Wenzel L,et al.Intraperitoneal cisplatin and paclitaxel in ovarian cancer[J].N Engl J Med,2006,354(1):34-43.DOI:10.1056/NEJMoa052985
[13]Koshy N,Quispe D,Shi R,et al.Cisplatin-gemcitabine therapy in metastatic breast cancer:Improved outcome in triple negative breast cancer patients compared to non-triple negative patients[J].Breast,2010,19(3):246-248.DOI:10.1016/j.breast.2010.02.003
[14]Zhu S,Pabla N,Tang C,et al.DNA damage response in cisplatin-induced nephrotoxicity[J].Arch Toxicol,2015,89(12):2197-2205.DOI:10.1007/s00204-015-1633-3
[15]Balakrishnan L,Stewart J,Polaczek P,et al.Acetylation of Dna2 endonuclease/helicase and flap endonuclease 1 by p300 promotes DNA stability by creating long flap intermediates[J].J Biol Chem,2010,285(7):4398-4404.DOI:10.1074/jbc.M109.086397
[16]马进举,陈彬,卞士柱.瓣状内切核酸酶1与肿瘤[J].生命的化学,2011,31(1):36-40.DOI:10.13488/j.smhx.2011.01.027
[17]Nikolova T,Christmann M,Kaina B.FEN1 is overexpressed in testis,lung and brain tumors[J].Anticancer Res,2009,29(7):2453-2459.
[18]Ye M X,Zhao Y L,Li Y,et al.Curcumin reverses cis-platin resistance and promotes human lung adenocarcinoma A549/DDP cell apoptosis through HIF-1αand caspase-3mechanisms[J].Phytomedicine,2012,19(8/9):779-787.DOI:10.1016/j.phymed.2012.03.005
[2]Ugur S,Ulu R,Dogukan A,et al.The renoprotective effect of curcumin in cisplatin-induced nephrotoxicity[J].Ren Fail,2015,37(2):332-336.DOI:10.3109/0886022X.2014.986005
[3]Wang J,Zhou L,Li Z,et al.YY1 suppresses FEN1 over-expression and drug resistance in breast cancer[J].BMC Cancer,2015,15:50.DOI:10.1186/s12885-015-1043-1
[4]Balakrishnan L,Bambara R A.Flap endonuclease 1[J].Annu Rev Biochem,2013,82:119-138.DOI:10.1146/annurev-biochem-072511-122603
[5]Abdel-Fatah T M,Russell R,Albarakati N,et al.Genomic and protein expression analysis reveals flap endonuclease 1(FEN1)as a key biomarker in breast and ovarian cancer[J].Mol Oncol,2014,8(7):1326-1338.DOI:10.1016/j.molonc.2014.04.009
[6]Chereddy K K,Coco R,Memvanga P B,et al.Combined effect of PLGA and curcumin on wound healing activity[J].J Control Release,2013,171(2):208-215.DOI:10.1016/j.jconrel.2013.07.015
[7]Hatcher H,Planalp R,Cho J,et al.Curcumin:from ancient medicine to current clinical trials[J].Cell Mol Life Sci,2008,65(11):1631-1652.DOI:10.1007/s00018-008-7452-4
[8]Chang Z,Xing J,Yu X.Curcumin induces osteosarcoma MG63 cells apoptosis via ROS/Cyto-C/Caspase-3 pathway[J].Tumour Biol,2014,35(1):753-758.DOI:10.1007/s13277-013-1102-7
[9]Lv Z D,Liu X P,Zhao W J,et al.Curcumin induces apoptosis in breast cancer cells and inhibits tumor growth in vitro and in vivo[J].Int J Clin Exp Pathol,2014,7(6):2818-2824.
[10]Chen B,Zhang Y,Wang Y,et al.Curcumin inhibits proliferation of breast cancer cells through Nrf2-mediated downregulation of Fen1 expression[J].J Steroid Biochem Mol Biol,2014,143:11-18.DOI:10.1016/j.jsbmb.2014.01.009
[11]Kelland L.The resurgence of platinum-based cancer chemotherapy[J].Nat Rev Cancer,2007,7(8):573-584.DOI:10.1038/nrc2167
[12]Armstrong D K,Bundy B,Wenzel L,et al.Intraperitoneal cisplatin and paclitaxel in ovarian cancer[J].N Engl J Med,2006,354(1):34-43.DOI:10.1056/NEJMoa052985
[13]Koshy N,Quispe D,Shi R,et al.Cisplatin-gemcitabine therapy in metastatic breast cancer:Improved outcome in triple negative breast cancer patients compared to non-triple negative patients[J].Breast,2010,19(3):246-248.DOI:10.1016/j.breast.2010.02.003
[14]Zhu S,Pabla N,Tang C,et al.DNA damage response in cisplatin-induced nephrotoxicity[J].Arch Toxicol,2015,89(12):2197-2205.DOI:10.1007/s00204-015-1633-3
[15]Balakrishnan L,Stewart J,Polaczek P,et al.Acetylation of Dna2 endonuclease/helicase and flap endonuclease 1 by p300 promotes DNA stability by creating long flap intermediates[J].J Biol Chem,2010,285(7):4398-4404.DOI:10.1074/jbc.M109.086397
[16]马进举,陈彬,卞士柱.瓣状内切核酸酶1与肿瘤[J].生命的化学,2011,31(1):36-40.DOI:10.13488/j.smhx.2011.01.027
[17]Nikolova T,Christmann M,Kaina B.FEN1 is overexpressed in testis,lung and brain tumors[J].Anticancer Res,2009,29(7):2453-2459.
[18]Ye M X,Zhao Y L,Li Y,et al.Curcumin reverses cis-platin resistance and promotes human lung adenocarcinoma A549/DDP cell apoptosis through HIF-1αand caspase-3mechanisms[J].Phytomedicine,2012,19(8/9):779-787.DOI:10.1016/j.phymed.2012.03.005