摘要
目的:观察辣椒碱对乳腺癌MDA-MB-231细胞裸鼠移植瘤生长的抑制作用及其机制。方法:通过皮下注射MDA-MB-231细胞的方法建立乳腺癌裸鼠移植瘤模型,待肿瘤体积达到约100 mm3,将裸鼠随机分为模型组和辣椒碱低、中、高剂量(5,10,20 mg·kg~(-1))组,每组5只。辣椒碱低、中、高剂量(5,10,20 mg·kg~(-1))组裸鼠分别腹腔注射相应剂量的辣椒碱,模型组注射等体积的磷酸缓冲盐溶液(PBS)溶剂,3 d/次,连续给药8次。每次给药前先检测裸鼠体质重和肿瘤体积,末次给药24 h后处死各组裸鼠,计算肿瘤体积,称肿瘤质量,并计算抑瘤率。应用免疫组化法和蛋白免疫印迹法(Western blot)检测辣椒碱治疗后各组肿瘤组织中高迁移率族蛋白B1(HMGB1)和Toll样受体4(TLR4)蛋白表达水平。结果:与模型组比较,辣椒碱低、中、高剂量(5,10,20 mg·kg~(-1))组裸鼠体质量无明显变化,但肿瘤体积显著缩小(P <0. 01),肿瘤质量显著减小(P <0. 01),抑瘤率显著增大(P <0. 01),且呈剂量依赖性;与模型组比较,辣椒碱低、中、高剂量(5,10,20 mg·kg~(-1))组肿瘤组织HMGB1和TLR4蛋白表达明显降低,且呈剂量依赖性。结论:辣椒碱对乳腺癌MDA-MB-231细胞裸鼠移植瘤生长具有明显的抑制作用,其机制可能与下调HMGB1和TLR4蛋白表达有关。
Objective: To investigate the inhibitory effect of capsaicin on the growth of breast cancer MDA-MB-231 cells transplanted tumour in nude mice and its possible molecular mechanism. Method:Transplanted tumor model of breast cancer MDA-MB-231 cells in nude mice were established. Then the tumorbearing mice were randomly divided into 4 groups: model group,and low,medium and high-dose capsaicin groups( 5,10,20 mg·kg~(-1)). Mice of low,medium and high-dose capsaicin groups( 5,10,20 mg·kg~(-1)) were intraperitoneally injected with the corresponding dose of capsaicin,and the model group was injected with the same volume of phosphate buffer saline( PBS),once every 3 days,for a total of 8 times in succession. Body weight of mice and transplantation tumor volume were measured before each injection of capsaicin. Mice of each group were put to death 24 h after the last administration,and then the tumor volume,mass and the tumor inhibitory rate were calculated. The protein expression levels of high mobility group box 1( HMGB1) and Toll-like receptors 4( TLR4) were measured by immunohistochemistry and Western blot. Result: No significant difference was observed between each group in body weight. However,compared with the model group,capsaicin( 5,10,20 mg·kg~(-1))remarkably inhibited the tumor volume and mass( P < 0. 05,P < 0. 01),and increased the tumor inhibitory rate( P < 0. 01) in a dose-dependent manner. Capsaicin( 5,10,20 mg·kg~(-1)) also markedly inhibited the protein expression levels of HMGB1 and TLR4( P < 0. 01). Conclusion: Capsaicin remarkably inhibits the growth of breast cancer MDA-MB-231 cells transplanted tumour in nude mice,and the possible mechanism may be related to the down-regulation of HMGB1 and TLR4 at the protein level.
引文
[1]陈万青,郑荣寿.中国女性乳腺癌发病死亡和生存状况[J].中国肿瘤临床,2015,42(13):668-674.
[2] MAO X L,ZHU H Y,LUO D H,et al. Capsaicin inhibits glycolysis in esophageal squamous cell carcinoma by regulating hexokinase 2 expression[J].Mol Med Rep,2018,17(4):6116-6121.
[3] Friedman J R,Nolan N A,Miles S L,et al. Anticancer activity of natural and synthetic capsaicin analogs[J]. J Pharmacol Exp Ther,2018,364(3):462-473.
[4] Diaz-Laviada I, Rodriguez-Henche N. The potential antitumor effect of capsaicin[J]. Prog Drug Res,2014,68:181-208.
[5]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.
[6]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.
[7]王振兴,孙中莉,王明杰,等.补阳还五汤对肺纤维化中HMGB1-RAGE信号通路的调控作用[J].中国实验方剂学杂志,2017,23(13):138-144.
[8] Sims G P,Rowe D C,Rietdijk S T,et al. HMGB1 and RAGE in inflammation and cancer[J]. Annu Rev Immunol,2010,28:367-388.
[9] YAN W,CHANG Y,LIANG X Y,et al. High-mobility group box 1 activates Caspase-1 and promotes hepatocellular carcinoma invasiveness and metastases[J]. Hepatology,2012,55(6):1863-1875.
[10] SUN S P,ZHANG W,CUI Z Q,et al. High mobility group box-1 and its clinical value in breast cancer[J].Onco Targets Ther,2015,8:413-419.
[11] KE S B,ZHOU F X,YANG H,et al. Downregulation of high mobility group box 1 modulates telomere homeostasis and increases the radiosensitivity of human breast cancer cells[J]. Int J Oncol,2015,46(3):1051-1058.
[12]陈茂剑,王丽,杨伟萍,等.沉默FBI-1基因对三阴性乳腺癌细胞MDA-MB-231增殖和凋亡的影响[J].生理学报,2018,70(5):497-503.
[13]张鹏程,陈美周,余方流,等.辣椒素对小鼠B16F10黑色素瘤抑制效应研究[J].中华肿瘤防治杂志,2015,22(14):1096-1099.
[14] Friedman J R, Nolan N A, Brown K C, et al.Anticancer activity of natural and synthetic capsaicin analogs[J]. J Pharmacol Exp Ther,2018,364(3):462-473.
[15] Chapa-Oliver A M,Mejia-Teniente L. Capsaicin:from plants to a cancer-suppressing agent[J]. Molecules,2016,21(8):E931.
[16] LIN Y T,WANG H C,Hsu Y C,et al. Capsaicin induces autophagy and apoptosis in human nasopharyngeal carcinoma cells by downregulating the PI3K/AKT/m TOR pathway[J]. Int J Mol Sci,2017,18(7):E1343.
[17] 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.
[18]李日飞,袁娜,冶冬阳,等.乳腺癌实验动物模型的研究进展[J].中国比较医学杂志,2018,28(2):113-118.
[19]刘华钢,刘丽敏,黄慧学,等.抗肿瘤中药动物模型及机制研究进展[J].现代生物医学进展,2008,8(7):1334-1337.
[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] Goodwin G H,Sanders C,Johns E W. A new group of chromatin-associated proteins with a high content of acidic and basic amino acids[J]. Eur J Biochem,1973,38(1):14-19.
[22] CHEN M J,HUANG W J,WANG C,et al. Highmobility group box 1 exacerbates CCl(4)-induced acute liver injury in mice[J]. Clin Immunol,2014,153(1):56-63.
[23] Shimizu S,Kouzaki H,Kato T,et al. HMGB1-TLR4signaling contributes to the secretion of interleukin 6 and interleukin 8 by nasal epithelial cells[J]. Am J Rhinol Allergy,2016,30(3):167-172.
[24] Kumari T,Kumar B. High-mobility group box 1 protein(HMGB1)gene polymorphisms and cancer susceptibility:a comprehensive Meta-analysis[J]. Clin Chim Acta,2018,483:170-182.
[25] KANG R,ZHANG Q H,Zeh H J,et al. HMGB1 in cancer:good,bad,or both?[J]. Clin Cancer Res,2013,19(15):4046-4057.
[26] WU K L,ZHANG H H,FU Y J,et al. TLR4/MyD88signaling determines the metastatic potential of breast cancer cells[J]. Mol Med Rep, 2018, 18(3):3411-3420.
[27] LV W, CHEN N, LIN Y L, et al. Macrophage migration inhibitory factor promotes breast cancer metastasis via activation of HMGB1/TLR4/NF kappa B axis[J]. Cancer Lett,2016,375(2):245-255.
[28] HUANG B F,Tzeng H E,CHEN P C,et al. HMGB1genetic polymorphisms are biomarkers for the development and progression of breast cancer[J]. Int J Med Sci,2018,15(6):580-586.
[29]许政旭,朱诗国,潘年松,等.黔产莪术油对直肠癌SW1463细胞株分泌Toll样受体及相关免疫因子的影响[J].中国实验方剂学杂志,2018,24(5):137-141.
[30] WANG X B,YU X M,WANG Q L,et al. Expression and clinical significance of SATB1 and TLR4 in breast cancer[J]. Oncol Lett,2017,14(3):3611-3615.
[31]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):95-103.
[32] Sootichote R, Thuwajit P, Singsuksawat E, et al.Compound A attenuates toll-like receptor 4-mediated paclitaxel resistance in breast cancer and melanoma through suppression of IL-8[J]. BMC Cancer,2018,18(1):231.
[33] WANG F P,LI L,LI J,et al. High mobility group box-1 promotes the proliferation and migration of hepatic stellate cells via TLR4-dependent signal pathways of PI3K/Akt and JNK[J]. PLo S One, 2013, 8(5):e64373.
[34] JIANG C X,QU X,KE H H,et al. Association between the HMGB1/TLR4 signaling pathway and the clinicopathological features of ovarian cancer[J]. Mol Med Rep,2018,18(3):3093-3098.
