辣椒碱通过下调SIRT1表达抑制乳腺癌MCF-7细胞的迁移和侵袭
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摘要
目的:观察辣椒碱对人乳腺癌MCF-7细胞迁移和侵袭的影响,并初步探讨其分子生物学机制。方法:设立辣椒碱(25,50,75μmol·L~(-1))组以及空白组,采用穿透小室(Transwell)迁移和侵袭实验分别检测辣椒碱(25,50,75μmol·L~(-1))干预乳腺癌MCF-7细胞24 h后和空白组细胞的迁移和侵袭能力;实时荧光定量聚合酶链式反应(Real-time PCR)检测辣椒碱(25,50,75μmol·L~(-1))干预乳腺癌MCF-7细胞24 h后和空白组细胞中沉默信息调节因子2同源蛋白1(SIRT1)和DNA聚合酶δ催化亚基p125的编码基因POLD1(POLD1) mRNA表达水平;蛋白免疫印迹法(Western blot)检测辣椒碱(25,50,75μmol·L~(-1))干预乳腺癌MCF-7细胞24 h后和空白组细胞中SIRT1和DNA聚合酶δ催化亚基p125(p125)的蛋白表达水平。结果:与空白组比较,辣椒碱(25,50,75μmol·L~(-1))干预乳腺癌MCF-7细胞24 h后,穿膜细胞数明显减少,迁移率和侵袭率显著降低,且呈浓度依赖效应(P <0. 01);与空白组比较,辣椒碱(25,50,75μmol·L~(-1))干预24 h能显著下调乳腺癌MCF-7细胞中SIRT1,POLD1 mRNA表达水平(P <0. 01);与空白组比较,辣椒碱(25,50,75μmol·L~(-1))干预24 h能显著降低乳腺癌MCF-7细胞中SIRT1和DNA聚合酶δ催化亚基p125的蛋白表达水平(P <0. 01)。结论:辣椒碱能显著抑制乳腺癌MCF-7细胞的迁移和侵袭能力,其机制可能与下调细胞中SIRT1 mRNA和蛋白的表达水平以及POLD1 mRNA和p125的蛋白表达水平有关。
Objective: To investigate the effect of capsaicin on the migration and invasion of human breast cancer MCF-7 cells and the underlying molecular mechanism. Method: Three capsaicin intervention groups of different concentrations( 25,50,75 μmol·L~(-1)) and a blank group were set up. After MCF-7 cells were treated with different concentrations of capsaicin( 25,50,75 μmol·L~(-1)) for 24 h,the cell migration and invasion abilities were assessed by Transwell migration and invasion assay,respectively. Meanwhile,the mRNA level of silent information regulator 2 homolog 1( SIRT1) and DNA polymerase δ catalytic subunit p125 encoding gene POLD1( POLD1) were detected by Real-time polymerase chain reaction( Real-time PCR). The protein levels of SIRT1 and DNA polymerase δ catalytic subunit p125( p125) were detected by Western blot. Result: Compared with the blank group,the number of transmembrane cells was significantly reduced,and the mobility was significantly decreased( P < 0. 01) in a dose-dependent manner after the intervention of capsaicin( 25,50,75 μmol·L~(-1)) in MCF-7 cells for 24 h. Capsaicin( 25,50,75 μmol·L~(-1)) significantly down-regulated the mRNA and protein expressions of SIRT1( P < 0. 01) after intervention of capsaicin( 25,50,75 μmol·L~(-1)) in MCF-7 cells for 24 h.Furthermore,capsaicin( 25,50,75 μmol·L~(-1)) also significantly down-regulated the mRNA expression of POLD1 and the protein expression of p125( P < 0. 01) after intervention of capsaicin( 25,50,75 μmol·L~(-1)) in MCF-7 cells for 24 h. Conclusion: Capsaicin remarkably inhibits the cell migration and invasion of breast cancer MCF-7 cells,and the possible mechanism may be related to the down-regulation of SIRT1 and POLD1 mRNA expression levels and SIRT1 and p125 protein expression levels.
Objective: To investigate the effect of capsaicin on the migration and invasion of human breast cancer MCF-7 cells and the underlying molecular mechanism. Method: Three capsaicin intervention groups of different concentrations( 25,50,75 μmol·L~(-1)) and a blank group were set up. After MCF-7 cells were treated with different concentrations of capsaicin( 25,50,75 μmol·L~(-1)) for 24 h,the cell migration and invasion abilities were assessed by Transwell migration and invasion assay,respectively. Meanwhile,the mRNA level of silent information regulator 2 homolog 1( SIRT1) and DNA polymerase δ catalytic subunit p125 encoding gene POLD1( POLD1) were detected by Real-time polymerase chain reaction( Real-time PCR). The protein levels of SIRT1 and DNA polymerase δ catalytic subunit p125( p125) were detected by Western blot. Result: Compared with the blank group,the number of transmembrane cells was significantly reduced,and the mobility was significantly decreased( P < 0. 01) in a dose-dependent manner after the intervention of capsaicin( 25,50,75 μmol·L~(-1)) in MCF-7 cells for 24 h. Capsaicin( 25,50,75 μmol·L~(-1)) significantly down-regulated the mRNA and protein expressions of SIRT1( P < 0. 01) after intervention of capsaicin( 25,50,75 μmol·L~(-1)) in MCF-7 cells for 24 h.Furthermore,capsaicin( 25,50,75 μmol·L~(-1)) also significantly down-regulated the mRNA expression of POLD1 and the protein expression of p125( P < 0. 01) after intervention of capsaicin( 25,50,75 μmol·L~(-1)) in MCF-7 cells for 24 h. Conclusion: Capsaicin remarkably inhibits the cell migration and invasion of breast cancer MCF-7 cells,and the possible mechanism may be related to the down-regulation of SIRT1 and POLD1 mRNA expression levels and SIRT1 and p125 protein expression levels.
引文
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[14]陈茂剑,杨伟萍,覃庆洪,等.辣椒碱对乳腺癌MCF-7细胞增殖及p21和FBI-1表达的影响[J].中国实验方剂学杂志,2018,24(22):102-106.
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[17] Clark R,Lee S H. Anticancer properties of capsaicin against human cancer[J]. Anticancer Res,2016,36(3):837.
[18] Chapa-Oliver A M,Mejía-Teniente L. Capsaicin:from plants to a cancer-suppressing agent[J]. Molecules,2016,21(8):931.
[19] CHANG H C,CHEN S T,Chien S Y,et al. Capsaicin may induce breast cancer cell death through apoptosisinducing factor involving mitochondrial dysfunction[J].Hum Exp Toxicol,2011,30(10):1657-1665.
[20] Thoennissen N H,O'Kelly J,LU D,et al. Capsaicin causes cell-cycle arrest and apoptosis in ER-positive and-negative breast cancer cells by modulating the EGFR/HER-2 pathway[J]. Oncogene,2009,29(2):285-296.
[21]李伯和,袁磊.辣椒碱对乳腺癌MDA-MB-231细胞迁移和侵袭的抑制作用及其机制[J].生理学报,2017,69(2):183-188.
[22] Chalkiadaki A,Guarente L. The multifaceted functions of sirtuins in cancer[J]. Nat Rev Cancer,2015,15(10):608-624.
[23] Hwang B J,Madabushi A,JIN J,et al. Histone/protein deacetylase SIRT1 is an anticancer therapeutic target[J]. Am J Cancer Res,2014,4(3):211-221.
[24] HU Q X,WANG G,PENG J P,et al. Knockdown of SIRT1 suppresses bladder cancer cell proliferation and migration and induces cell cycle arrest and antioxidant response through FOXO3a-mediated pathways[J].Biomed Res Int,2017,2017(9):3781904.
[25] XU Y F,QIN Q H,CHEN R S,et al. SIRT1 promotes proliferation,migration,and invasion of breast cancer cell line MCF-7 by upregulating DNA polymerase delta1(POLD1)[J]. Biochem Biophys Res Commun,2018,502(3):351-357.
[26] Nicolas E,Golemis E A,Arora S. POLD1:Central mediator of DNA replication and repair,and implication in cancer and other pathologies[J]. Gene,2016,590(1):128-141.
[27]陈茂剑,蒋玮,覃庆洪,等.辣椒碱抗肿瘤作用分子机制[J].中国实验方剂学杂志,2019,25(7):100-108.
[28]欧贤红,廖柳凤,刘华钢,等. POLD1基因在原发性肝癌中的表达及其意义[J].世界华人消化杂志,2011,19(2):151-155.
[29] Sanefuji K,Taketomi A,Iguchi T,et al. Significance of DNA polymerase delta catalytic subunit p125 induced by mutant p53 in the invasive potential of human hepatocellular carcinoma[J]. Oncology,2010,79(3/4):229-237.
[2] Nho K J,Chun J M,Kim D S,et al. Ampelopsis japonica ethanol extract suppresses migration and invasion in human MDA-MB-231 breast cancer cells[J]. Mol Med Rep,2015,11(5):3722-3728.
[3] Chapa-Oliver A M,Mejia-Teniente L. Capsaicin:from plants to a cancer-suppressing agent[J]. Molecules,2016,21(8):931.
[4] Clark R,Lee S H. Anticancer properties of capsaicin against human cancer[J]. Anticancer Res,2016,36(3):837-843.
[5]肖潺潺,陈茂剑,梅凡彪,等.辣椒碱对肝癌SMMC-7721细胞迁移和侵袭的影响及机制[J].中国实验方剂学杂志,2018,24(18):124-129.
[6] Karbasforooshan H,Roohbakhsh A,Karimi G. SIRT1and microRNAs:the role in breast,lung and prostate cancers[J]. Exp Cell Res,2018,367(1):1-6.
[7] Sarma P,Bag I,Ramaiah M J,et al. Bisindole-PBD regulates breast cancer cell proliferation via SIRT-p53axis[J]. Cancer Biol Ther, 2015, 16(10):1486-1501.
[8] SHU Y,REN L G,XIE B,et al. MiR-204 enhances mitochondrial apoptosis in doxorubicin-treated prostate cancer cells by targeting SIRT1/p53 pathway[J].Oncotarget,2017,8(57):97313-97322.
[9] LIN Z H,FANG D Y. The roles of SIRT1 in cancer[J]. Genes Cancer,2013,4(3/4):97-104.
[10] Chung S Y,Jung Y Y,Park I A,et al. Oncogenic role of SIRT1 associated with tumor invasion,lymph node metastasis, and poor disease-free survival in triple negative breast cancer[J]. Clin Exp Metastasis,2016,33(2):179-185.
[11] TAN J,LIU Y,Maimaiti Y,et al. Combination of SIRT1 and Src overexpression suggests poor prognosis in luminal breast cancer[J]. Onco Targets Ther,2018,11:2051-2061.
[12] JIN X X,WEI Y Z,XU F,et al. SIRT1 promotes formation of breast cancer through modulating Akt activity[J]. J Cancer,2018,9(11):2012-2023.
[13] Chung Y R,Kim H,Park S Y,et al. Distinctive role of SIRT1 expression on tumor invasion and metastasis in breast cancer by molecular subtype[J]. Hum Pathol,2015,46(7):1027-1035.
[14]陈茂剑,杨伟萍,覃庆洪,等.辣椒碱对乳腺癌MCF-7细胞增殖及p21和FBI-1表达的影响[J].中国实验方剂学杂志,2018,24(22):102-106.
[15] Aggarwal B B,Shishodia S. Molecular targets of dietary agents for prevention and therapy of cancer[J].Biochem Pharmacol,2006,71(10):1397-1421.
[16]贺栋业,李晓宇,王丽丽,等.金花茶化学成分及药理作用研究进展[J].中国实验方剂学杂志,2016,22(3):231-234.
[17] Clark R,Lee S H. Anticancer properties of capsaicin against human cancer[J]. Anticancer Res,2016,36(3):837.
[18] Chapa-Oliver A M,Mejía-Teniente L. Capsaicin:from plants to a cancer-suppressing agent[J]. Molecules,2016,21(8):931.
[19] CHANG H C,CHEN S T,Chien S Y,et al. Capsaicin may induce breast cancer cell death through apoptosisinducing factor involving mitochondrial dysfunction[J].Hum Exp Toxicol,2011,30(10):1657-1665.
[20] Thoennissen N H,O'Kelly J,LU D,et al. Capsaicin causes cell-cycle arrest and apoptosis in ER-positive and-negative breast cancer cells by modulating the EGFR/HER-2 pathway[J]. Oncogene,2009,29(2):285-296.
[21]李伯和,袁磊.辣椒碱对乳腺癌MDA-MB-231细胞迁移和侵袭的抑制作用及其机制[J].生理学报,2017,69(2):183-188.
[22] Chalkiadaki A,Guarente L. The multifaceted functions of sirtuins in cancer[J]. Nat Rev Cancer,2015,15(10):608-624.
[23] Hwang B J,Madabushi A,JIN J,et al. Histone/protein deacetylase SIRT1 is an anticancer therapeutic target[J]. Am J Cancer Res,2014,4(3):211-221.
[24] HU Q X,WANG G,PENG J P,et al. Knockdown of SIRT1 suppresses bladder cancer cell proliferation and migration and induces cell cycle arrest and antioxidant response through FOXO3a-mediated pathways[J].Biomed Res Int,2017,2017(9):3781904.
[25] XU Y F,QIN Q H,CHEN R S,et al. SIRT1 promotes proliferation,migration,and invasion of breast cancer cell line MCF-7 by upregulating DNA polymerase delta1(POLD1)[J]. Biochem Biophys Res Commun,2018,502(3):351-357.
[26] Nicolas E,Golemis E A,Arora S. POLD1:Central mediator of DNA replication and repair,and implication in cancer and other pathologies[J]. Gene,2016,590(1):128-141.
[27]陈茂剑,蒋玮,覃庆洪,等.辣椒碱抗肿瘤作用分子机制[J].中国实验方剂学杂志,2019,25(7):100-108.
[28]欧贤红,廖柳凤,刘华钢,等. POLD1基因在原发性肝癌中的表达及其意义[J].世界华人消化杂志,2011,19(2):151-155.
[29] Sanefuji K,Taketomi A,Iguchi T,et al. Significance of DNA polymerase delta catalytic subunit p125 induced by mutant p53 in the invasive potential of human hepatocellular carcinoma[J]. Oncology,2010,79(3/4):229-237.