Chemerin在乳腺癌小鼠外周血和乳腺组织中的表达及其对乳腺癌细胞增殖和迁移的影响
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摘要
目的 Chemerin是否与乳腺癌的发生与发展过程有关,目前报道较少。文章旨在探讨Chemerin在乳腺癌小鼠中的表达及其对乳腺癌细胞增殖与迁移的影响。方法将30只Balb/c小鼠随机分成正常小鼠和荷瘤小鼠,各15只。4T1乳腺癌细胞株接种Balb/c小鼠构建乳腺癌模型,通过ELISA和Western blot检测成瘤小鼠外周血和乳腺组织中Chemerin蛋白的表达量,分析Chemerin蛋白的表达水平与乳腺癌的关系;采用不同浓度的Chemerin重组蛋白和Chemerin中和抗体处理人乳腺癌细胞系;实验设置对照组(只含高糖1640或DMEM培养基,无细胞)、Chemerin重组蛋白组(100μg/L,无血清培养基进行稀释)以及Chemerin中和抗体组(100μg/L,无血清培养基进行稀释),通过MTT和划痕实验检测Chemerin对癌细胞增殖和迁移的影响。结果 ELISA结果显示,与正常小鼠相比,荷瘤小鼠的外周血表达量均明显增高[(116.5±18.6)pg/mL vs(1011.0±8.8)pg/mL,P<0.05],乳腺组织中Chemerin蛋白的表达量亦增高[(189.2±7.3)pg/mL vs(554.2±49.1)pg/mL,P<0.05]。Western blot结果显示,荷瘤小鼠的乳腺组织中Chemerin蛋白的表达量(1.561±0.133)亦明显高于正常小鼠(0.910±0.035),差异就有统计学意义(P<0.05)。MTT法和划痕实验结果显示,Chemerin重组蛋白组MCF-7、MDA-MB-231细胞增殖率[(0.715±0.027)%、(1.617±0.065)%]较对照组[(0.429±0.024)%、(1.151±0.102)%]明显升高(P<0.05), MDA-MB-231细胞的迁移率[(29.870±5.730)%]较对照组[(19.610±2.485)%]明显增加(P<0.01);Chemerin中和抗体组细胞增殖率[0.279±0.008)%、(0.941±0.086)%、(9.473±0.843)%]较对照组降低(P<0.05)。结论 Chemerin呈浓度梯度式促进乳腺癌细胞的增殖和迁移,从而促进乳腺癌的发生发展。
Objective At present,there are few reports about the relationship between Chemerin and the occurrence and development of breast cancer. The aim of this study is to investigate Chemerin expression in breast cancer mice and its effect on proliferation and migration of breast cancer cells.Methods 30 Balb/c mice were randomly divided into two groups(Normal mice 15,Tumor-bearing mice 15). The 4 T1 breast cancer cells were inoculated to construct breast cancer mice model. The expressions of Chemerin in peripheral blood and breast cancer tissue were detected by ELISA and Western blot,respectively. The relationship between Chemerin expression and breast cancer was analyzed. Breast cancer cells MCF-7 and MDA-MB-231 were treated with different concentrations of recombinant Chemerin protein and Chemerin neutralizing antibody.Three groups were set up in the experiment,including control group(1640 or DMEM culture medium,no cells),recombinant Chemerin protein group(100 μg/L,no dilution by serum-free medium)and Chemerin neutralizing antibody group(100 μg/L,no dilution by serum-free medium). The effects of Chemerin on cell proliferation and migration were detected by MTT assay and wound healing assay respectively.Results The expressions of Chemerin in peripheral blood and mammary gland of tumor bearing mice were significantly higher than that in normal mice(both P<0.05). The scratch mobility and MTT absorbance(OD)values of breast cancer cells treated with recombinant chemerin protein increased significantly with the increase of protein concentration(P<0.05).However,they significantly decreased after cells treated with chemerin neutralizing antibody(P<0.05).Conclusion Chemerin could promote the proliferation and migration of breast cancer cells in a concentration-dependent manner,thereby promotinge the occurrence and development of breast cancer.
Objective At present,there are few reports about the relationship between Chemerin and the occurrence and development of breast cancer. The aim of this study is to investigate Chemerin expression in breast cancer mice and its effect on proliferation and migration of breast cancer cells.Methods 30 Balb/c mice were randomly divided into two groups(Normal mice 15,Tumor-bearing mice 15). The 4 T1 breast cancer cells were inoculated to construct breast cancer mice model. The expressions of Chemerin in peripheral blood and breast cancer tissue were detected by ELISA and Western blot,respectively. The relationship between Chemerin expression and breast cancer was analyzed. Breast cancer cells MCF-7 and MDA-MB-231 were treated with different concentrations of recombinant Chemerin protein and Chemerin neutralizing antibody.Three groups were set up in the experiment,including control group(1640 or DMEM culture medium,no cells),recombinant Chemerin protein group(100 μg/L,no dilution by serum-free medium)and Chemerin neutralizing antibody group(100 μg/L,no dilution by serum-free medium). The effects of Chemerin on cell proliferation and migration were detected by MTT assay and wound healing assay respectively.Results The expressions of Chemerin in peripheral blood and mammary gland of tumor bearing mice were significantly higher than that in normal mice(both P<0.05). The scratch mobility and MTT absorbance(OD)values of breast cancer cells treated with recombinant chemerin protein increased significantly with the increase of protein concentration(P<0.05).However,they significantly decreased after cells treated with chemerin neutralizing antibody(P<0.05).Conclusion Chemerin could promote the proliferation and migration of breast cancer cells in a concentration-dependent manner,thereby promotinge the occurrence and development of breast cancer.
引文
[1] Perumalsamy S,Aqilah Mohd Zin NA,Widodo RT,et al. Chemokine like receptor-1(CMKLR-1)receptor:a potential therapeutic target in management of chemerin induced type 2 diabetes mellitus and cancer[J]. Curr Pharm Des,2017,23(25):3689-3698.
[2]黄泽锋,谢鑫. Chemerin通过核因子κB介导的炎症反应诱导C2C12细胞产生胰岛素抵抗[J].细胞与分子免疫学杂志,2015,31(6):725-729.
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[5] Coban M,Tasli L,Turgut S,et al. Association of adipokines,insulin resistance,hypertension and dyslipidemia in patients with psoriasis vulgaris[J]. Ann Dermatol,2016,28(1):74-79.
[6]林小晶,王晓慧.炎症因子chemerin在肥胖及其相关疾病中的作用及运动对chemerin调控的研究进展[J].生理科学进展,2017,48(5):383-387.
[7] Zhang J,Zhou J,Tang X,et al. Reduced expression of chemerin is associated with poor clinical outcome in acute myeloid leukemia[J]. Oncotarget,2017,8(54):92536-92544.
[8] Liu-Chittenden Y,Patel D,Gaskins K,et al. Serum RARRES2is a prognostic marker in patients with adrenocortical carcinoma[J]. J Clin Endocrinol Metabol,2016,101(9):3345-3352.
[9] Sarmadi P,Tunali G,Esendagli-Yilmaz G,et al. CRAM-A indicates IFN-gamma-associated inflammatory response in breast cancer[J]. Mol Immunol,2015,68(2):692-698.
[10] Krawczyk KM,Nilsson H,Allaoui R,et al. Papillary renal cell carcinoma-derived chemerin,IL-8,and CXCL16 promote monocyte recruitment and differentiation into foam-cell macrophages[J]. Lab Invest,2017,97(11):1296-1305.
[11]高玮男,张雪双,孙连坤,等. SAS诱导氧化应激时人卵巢癌顺铂敏感细胞与耐药细胞死亡方式的差异[J].中国实验诊断学,2018,22(4):690-694.
[12] Wang H,Luo Q,Feng X,et al. NLRP3 promotes tumor growth and metastasis in human oral squamous cell carcinoma[J]. BMC Cancer,2018,18(1):500.
[13]黄美华,朱晨宇,商远方.华蟾素对宫颈癌细胞增殖的影响[J].现代肿瘤医学,2015,23(13):1802-1804.
[14] Huang T,Gong WH,Li XC,et al. Efficient killing effect of osteosarcoma cells by cinobufacini and cisplatin in combination[J]. Asian Pac J Cancer Prev,2012,13(6):2847-2851.
[15] Zhang Q,Ding L,Liang X,et al. Comparison of pathological characteristics between self-detected and screen-detected invasive breast cancers in Chinese women:a retrospective study[J].PeerJ,2018,6:e4567.
[16]师金,梁迪,李道娟,等.全球女性乳腺癌流行情况研究[J].中国肿瘤,2017,26(9):683-690.
[17]朱文斌,温宪春,于海涛,等. PCDHA13基因启动子甲基化与乳腺癌的相关性[J].医学研究生学报,2018,31(10):1026-1032.
[18]王璐璐,曾慧娟,王婧洁,等.长链非编码RNA小核仁RNA宿主基因3在乳腺癌组织中的表达及其临床意义[J].医学研究生学报,2018,31(11):1180-1183.
[19]屈晓冰,高洁,赵水平.脂肪组织和脂肪细胞因子的研究进展[J].国际病理科学与临床杂志,2008,28(4):353-357.
[20] Lee YC,Chen YJ,Wu CC,et al. Resistin expression in breast cancer tissue as a marker of prognosis and hormone therapy stratification[J]. Gynecol Oncol,2012,125(3):742-750.
[21]刘一锋,蒋玉林,张志乾,等.瘦素通过下调FBP1基因表达促进人乳腺癌BT-474和MDA-MB-231细胞增殖并抑制细胞凋亡[J].肿瘤,2017,37(6):556-567.
[22]刘文慧,张秋瑾,周心怡,等.瘦素促进人乳腺癌细胞株MCF-7增殖的机制研究[J].实用医学杂志,2017,33(4):27-30.
[23] Goralski KB,McCarthy TC,Hanniman EA,et al. Chemerin,a novel adipokine that regulates adipogenesis and adipocyte metabolism[J]. J Biol Chem,2007,282(38):28175-28188.
[24] Kumar JD,Kandola S,Tiszlavicz L,et al. The role of chemerin and ChemR23 in stimulating the invasion of squamous oesophageal cancer cells[J]. Br J Cancer,2016,114(10):1152-1159.
[25] Zhang J,Jin HC,Zhu AK,et al. Prognostic significance of plasma chemerin levels in patients with gastric cancer[J]. Peptides,2014,61:7-11.
[26]曲晓翰,韩立波,王绍成,等.肺癌患者外周血中Chemerin的测定及临床意义[J].中国肺癌杂志,2009,12(11):1174-1177.
[27] Eichelmann F,Schulze MB,Wittenbecher C,et al. Association of Chemerin Plasma Concentration With Risk of Colorectal Cancer[J]. JAMA Netw Open,2019,2(3):e190896.
[28] Alkady MM,Abdel-Messeih PL,Nosseir NM. Assessment of Serum Levels of the Adipocytokine Chemerin in Colorectal Cancer Patients[J]. J Med Biochem,2018,37(3):313-319.
[29] Farsam V,Basu A,Gatzka M,et al. Senescent fibroblast-derived Chemerin promotes squamous cell carcinoma migration[J]. Oncotarget,2016,7(50):83554-83569.
[2]黄泽锋,谢鑫. Chemerin通过核因子κB介导的炎症反应诱导C2C12细胞产生胰岛素抵抗[J].细胞与分子免疫学杂志,2015,31(6):725-729.
[3]闫丽丽,吕骥,贾红玉,等.血清chemerin水平与2型糖尿病、非酒精性脂肪性肝病的关系[J].肝脏,2018,23(3):239-241.
[4] Lima PDA,Nivet AL,Wang Q,et al. Polycystic ovary syndrome:possible involvement of androgen-induced,chemerinmediated ovarian recruitment of monocytes/macrophages[J]. Biol Reprod,2018,99(4):838-852.
[5] Coban M,Tasli L,Turgut S,et al. Association of adipokines,insulin resistance,hypertension and dyslipidemia in patients with psoriasis vulgaris[J]. Ann Dermatol,2016,28(1):74-79.
[6]林小晶,王晓慧.炎症因子chemerin在肥胖及其相关疾病中的作用及运动对chemerin调控的研究进展[J].生理科学进展,2017,48(5):383-387.
[7] Zhang J,Zhou J,Tang X,et al. Reduced expression of chemerin is associated with poor clinical outcome in acute myeloid leukemia[J]. Oncotarget,2017,8(54):92536-92544.
[8] Liu-Chittenden Y,Patel D,Gaskins K,et al. Serum RARRES2is a prognostic marker in patients with adrenocortical carcinoma[J]. J Clin Endocrinol Metabol,2016,101(9):3345-3352.
[9] Sarmadi P,Tunali G,Esendagli-Yilmaz G,et al. CRAM-A indicates IFN-gamma-associated inflammatory response in breast cancer[J]. Mol Immunol,2015,68(2):692-698.
[10] Krawczyk KM,Nilsson H,Allaoui R,et al. Papillary renal cell carcinoma-derived chemerin,IL-8,and CXCL16 promote monocyte recruitment and differentiation into foam-cell macrophages[J]. Lab Invest,2017,97(11):1296-1305.
[11]高玮男,张雪双,孙连坤,等. SAS诱导氧化应激时人卵巢癌顺铂敏感细胞与耐药细胞死亡方式的差异[J].中国实验诊断学,2018,22(4):690-694.
[12] Wang H,Luo Q,Feng X,et al. NLRP3 promotes tumor growth and metastasis in human oral squamous cell carcinoma[J]. BMC Cancer,2018,18(1):500.
[13]黄美华,朱晨宇,商远方.华蟾素对宫颈癌细胞增殖的影响[J].现代肿瘤医学,2015,23(13):1802-1804.
[14] Huang T,Gong WH,Li XC,et al. Efficient killing effect of osteosarcoma cells by cinobufacini and cisplatin in combination[J]. Asian Pac J Cancer Prev,2012,13(6):2847-2851.
[15] Zhang Q,Ding L,Liang X,et al. Comparison of pathological characteristics between self-detected and screen-detected invasive breast cancers in Chinese women:a retrospective study[J].PeerJ,2018,6:e4567.
[16]师金,梁迪,李道娟,等.全球女性乳腺癌流行情况研究[J].中国肿瘤,2017,26(9):683-690.
[17]朱文斌,温宪春,于海涛,等. PCDHA13基因启动子甲基化与乳腺癌的相关性[J].医学研究生学报,2018,31(10):1026-1032.
[18]王璐璐,曾慧娟,王婧洁,等.长链非编码RNA小核仁RNA宿主基因3在乳腺癌组织中的表达及其临床意义[J].医学研究生学报,2018,31(11):1180-1183.
[19]屈晓冰,高洁,赵水平.脂肪组织和脂肪细胞因子的研究进展[J].国际病理科学与临床杂志,2008,28(4):353-357.
[20] Lee YC,Chen YJ,Wu CC,et al. Resistin expression in breast cancer tissue as a marker of prognosis and hormone therapy stratification[J]. Gynecol Oncol,2012,125(3):742-750.
[21]刘一锋,蒋玉林,张志乾,等.瘦素通过下调FBP1基因表达促进人乳腺癌BT-474和MDA-MB-231细胞增殖并抑制细胞凋亡[J].肿瘤,2017,37(6):556-567.
[22]刘文慧,张秋瑾,周心怡,等.瘦素促进人乳腺癌细胞株MCF-7增殖的机制研究[J].实用医学杂志,2017,33(4):27-30.
[23] Goralski KB,McCarthy TC,Hanniman EA,et al. Chemerin,a novel adipokine that regulates adipogenesis and adipocyte metabolism[J]. J Biol Chem,2007,282(38):28175-28188.
[24] Kumar JD,Kandola S,Tiszlavicz L,et al. The role of chemerin and ChemR23 in stimulating the invasion of squamous oesophageal cancer cells[J]. Br J Cancer,2016,114(10):1152-1159.
[25] Zhang J,Jin HC,Zhu AK,et al. Prognostic significance of plasma chemerin levels in patients with gastric cancer[J]. Peptides,2014,61:7-11.
[26]曲晓翰,韩立波,王绍成,等.肺癌患者外周血中Chemerin的测定及临床意义[J].中国肺癌杂志,2009,12(11):1174-1177.
[27] Eichelmann F,Schulze MB,Wittenbecher C,et al. Association of Chemerin Plasma Concentration With Risk of Colorectal Cancer[J]. JAMA Netw Open,2019,2(3):e190896.
[28] Alkady MM,Abdel-Messeih PL,Nosseir NM. Assessment of Serum Levels of the Adipocytokine Chemerin in Colorectal Cancer Patients[J]. J Med Biochem,2018,37(3):313-319.
[29] Farsam V,Basu A,Gatzka M,et al. Senescent fibroblast-derived Chemerin promotes squamous cell carcinoma migration[J]. Oncotarget,2016,7(50):83554-83569.
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