Please wait a minute...
Journal of ZheJiang University(Medical Science)  2017, Vol. 46 Issue (4): 357-363    DOI: 10.3785/j.issn.1008-9292.2017.08.03
    
CXC chemokine receptor 4 regulates breast cancer cell cycle through S phase kinase associated protein 2
WANG Haifeng1, CHEN Tiantian1, WANG Yueyue1, LI Yu1, ZHANG Lingyu1, DING Yongxing2, CHEN Sulian3, WANG Wenrui4, YANG Qingling3, CHEN Changjie3
1. Clinical Testing and Diagnosis Center, Bengbu Medical College, Bengbu 233000, China;
2. Department of Surgical Oncology, Bengbu Third People's Hospital, Bengbu 233000, China;
3. Department of Biochemistry & Molecular Biology, Bengbu Medical College, Bengbu 233000, China;
4. Department of Biotechnology, Bengbu Medical College, Bengbu 233000, China
Download:   PDF(3532KB)
Export: BibTeX | EndNote (RIS)      

Abstract  

Objective:To investigate the effect of CXC chemokine receptor 4 (CXCR4) on cell cycle of breast cancer and its molecular mechanisms. Methods:The expression of CXCR4 and S phase kinase associated protein 2 (Skp2) was detected by real-time fluorescence quantitative PCR (fqRT-PCR) and Western blot in breast cancer cells. The expression of signal proteins and the downstream genes of Skp2 was detected by Western blot. The effect of CXCR4, PI3K/Akt pathway inhibitor LY294002 and ERK pathway inhibitor U0126 on cell cycle of breast cancer was detected by propidium iodide staining. Results:Skp2 was significantly down-regulated in CXCR4-downregulated cells and up-regulated in CXCR4-upregulated cells. CXCR4 also regulated the expression of Skp2 and other downstream genes by signaling protein. The proportion of cells in G0/G1 phase increased and that in S phase declined in CXCR4-downregulated cell, and the effect was more significant when combined with the use of LY294002 or U0126. Conclusion:CXCR4 can affect cell cycle and inhibit the proliferation of breast cancer cells by regulating Skp2 gene expression through PI3K/Akt and ERK signaling pathway.



Key wordsBreast neoplasms/physiopathology      Receptors, CXCR4/biosynthesis      Receptors, CXCR4/genetics      S-phase kinase-associated proteins/metabolism      Cell line, tumor/metabolism      Cell proliferation      Cell cycle     
Received: 16 February 2017      Published: 25 August 2017
CLC:  R737.9  
Cite this article:

WANG Haifeng, CHEN Tiantian, WANG Yueyue, LI Yu, ZHANG Lingyu, DING Yongxing, CHEN Sulian, WANG Wenrui, YANG Qingling, CHEN Changjie. CXC chemokine receptor 4 regulates breast cancer cell cycle through S phase kinase associated protein 2. Journal of ZheJiang University(Medical Science), 2017, 46(4): 357-363.

URL:

http://www.zjujournals.com/xueshu/med/10.3785/j.issn.1008-9292.2017.08.03     OR     http://www.zjujournals.com/xueshu/med/Y2017/V46/I4/357


CXC趋化因子受体4通过S期激酶相关蛋白2调控乳腺癌细胞周期的机制

目的:探讨CXC趋化因子受体4(CXCR4)通过PI3K/Akt和ERK信号通路调控S期激酶相关蛋白2(Skp2)的表达,进而影响乳腺癌细胞周期的机制。方法:利用干扰及过表达技术下调或上调CXCR4的表达后,通过实时定量PCR和蛋白质印迹法检测CXCR4与Skp2调控的关联性;蛋白质印迹法检测CXCR4干扰及过表达后对信号蛋白及Skp2下游相关基因表达的影响;通过碘化丙啶(PI)染色法检测CXCR4、PI3K/Akt通路抑制剂LY294002及ERK通路抑制剂U0126对乳腺癌细胞周期的影响。结果:干扰CXCR4后,Skp2表达下调;过表达CXCR4后,Skp2表达上调。CXCR4可通过对信号蛋白的调控影响Skp2及Skp2下游相关基因的表达。干扰CXCR4后,G0/G1期细胞比例增加,S期细胞比例相应减少,CXCR4与LY294002及U0126联合作用对细胞周期的阻断更加明显。结论:CXCR4能够通过对信号蛋白PI3K/Akt及ERK的调控,影响Skp2及Skp2下游相关基因的表达,阻断CXCR4/Akt/Skp2或CXCR4/ERK/Skp2信号通路后可有效诱导细胞周期阻滞,从而抑制乳腺癌细胞的增殖。


关键词: 乳腺肿瘤/病理生理学,  受体,  CXCR4/生物合成,  受体,  CXCR4/遗传学,  S期激酶相关蛋白质类/代谢,  细胞系,  肿瘤/代谢,  细胞增殖,  细胞周期 

[1] CONLEY-LACOMB M K, SALIGANAN A, KANDAGATLA P, et al. PTEN loss mediated Akt activation promotes prostate tumor growth and metastasis via CXCL12/CXCR4 signaling[J]. Mol Cancer,2013,12(1):85.
[2] FENG B, LI K, ZHONG H, et al. RhoE promotes metastasis in gastric cancer through a mechanism dependent on enhanced expression of CXCR4[J/OL]. PLoS One,2013,8(11):e81709.
[3] LIAO Y X, ZHOU C H, ZENG H, et al. The role of the CXCL12-CXCR4/CXCR7 axis in the progression and metastasis of bone sarcomas(review)[J]. Int J Mol Med,2013,32(6):1239-1246.
[4] YANG P, LIANG S X, HUANG W H, et al. Aberrant expression of CXCR4 significantly contributes to metastasis and predicts poor clinical outcome in breast cancer[J]. Curr Mol Med,2014,14(1):174-184.
[5] YANG Q, ZHANG F, DING Y,et al. Antitumour activity of the recombination polypeptide GST-NT21MP is mediated by inhibition of CXCR4 pathway in breast cancer[J]. Br J Cancer,2014,110(5):1288-1297.
[6] SONODA H, INOUE H, OGAWA K, et al.Significance of skp2 expression in primary breast cancer[J]. Clin Cancer Res,2006,12(4):1215-1220.
[7] LU Z, HUNTER T. Ubiquitylation and proteasomal degradation of the p21Cip1, p27Kip1 and p57Kip2 CDK inhibitors[J]. Cell Cycle,2010,9(12):2342-2352.
[8] YANG QL, ZHANG LY, WANG HF, et al. The N-terminal polypeptide derived from viral macrophage inflammatory protein Ⅱ reverses breast cancer epithelial-omesenchymal transition via a PDGFRa-dependent mechanism[J]. Oncotarge,2017,8(23):37448-37463.
[9] CHATTERJEE S, BEHNAM AZAD B, NIMMAGADDA S. The intricate role of CXCR4 in cancer[J]. Adv Cancer Res,2014,124:31-82.
[10] BAJETTO A, BARBIERI F, PATTAROZZI A, et al. CXCR4 and SDF1 expression in human meningiomas:a proliferative role in tumoral meningothelial cells in vitro[J]. Neuro Oncol,2007,9(1):3-11.
[11] ZHENG Q, SHUAI X, YE Y, et al. The role of polymorphisms of stromal derived factor-1 and CXC receptor 4 in acute myeloid leukemia and leukemia cell dissemination[J]. Gene,2016,588(2):103-108.
[12] 袁宏,冯虎,家秀秀,等.CXCR4 shRNA对U87胶质瘤细胞周期和凋亡的影响[J].中国热带医学,2015,15(1):25-27. YUAN Hong, FENG Hu, JIA Xiuxiu, et al. Effects of CXCR4 shRNA on cell cycle and apoptosis of U87 glioma[J]. China Tropical Medicine,2015,15(1):25-27. (in Chinese)
[13] 王艳,刘晓日,谭艳芳,等.RNA干扰沉默CXCR4基因对Jurkat细胞周期和凋亡的影响[J].中国实验血液学杂志,2010,18(3):625-628. WANG Yan, LIU Xiaori, TAN Yanfang, et al. Effects of CXCR4 silence induced by RNA interference on cell cycle distribution and apoptosis of jurkat cells[J]. Journal of Experimental Hematology,2010,18(3):625-628. (in Chinese)
[14] YANG Q, WU H, WANG H, et al. N-terminal polypeptide derived from vMIP-Ⅱ exerts its antitumor activity by inhibiting the CXCR4 pathway in human glioma[J]. Int J Oncol,2017,50(4):1160-1174.
[15] DEMETRICK D J, ZHANG H, BEACH D H. Chromosomal mapping of the genes for the human CDK2/cyclin A-associated proteins p19(SKP1A and SKP1B) and p45(SKP2)[J]. Cytogenet Cell Genet,1996,73(1-2):104-107.
[16] ZHANG H, KOBAYASHI R, GALAKTIONOV K, et al. p19Skp1 and p45Skp2 are essential elements of the cyclin A-CDK2 S phase kinase[J]. Cell,1995,82(6):915-925.
[17] CHAN C H, MORROW J K, ZHANG S, et al. Skp2:a dream target in the coming age of cancer therapy[J]. Cell Cycle,2014,13(5):679-680.
[18] PATERAS I S, APOSTOLOPOULOU K, KOUTSAMI M, et al. Downregulation of the KIP family members p27(KIP1) and p57(KIP2) by SKP2 and the role of methylation in p57(KIP2) inactivation in nonsmall cell lung cancer[J]. Int J Cancer,2006,119(11):2546-2556.
[19] FRESCAS M, PAGANO M. Deregulated proteolysis by the F-box proteins SKP2 and beta-TrCP:tipping the scales of cancer[J]. Nat Rev Cancer,2008,8(6):438-449.
[20] ZHOU W, SRINIVASAN S, NAWAZ Z, et al. ERα, SKP2 and E2F-1 form a feed forward loop driving late ERα targets and G1 cell cycle progression[J]. Oncogene,2014,33(18):2341-2353.
[21] WANG G, CHAN C H, GAO Y, et al. Novel roles of Skp2 E3 ligase in cellular senescence, cancer progression, and metastasis[J]. Chin J Cancer,2012,31(4):169-177.
[22] MITREA D M, YOON M K, OU L, et al. Disorder-function relationships for the cell cycle regulatory proteins p21 and p27[J]. Biol Chem,2012,393(4):259-274.
[23] 赵君,杨春鹿,赵苏英.肺癌中细胞S期激酶相关蛋白2的表达及其对预后的影响[J].中华肿瘤杂志,2007,29(4):289-292. ZHAO Jun, YANG Chunlu, ZHAO Suying. Expression of Skp2 protein in lung carcinoma and its implication for prognosis[J]. Chinese Journal of Oncology,2007,29(4):289-292. (in Chinese)
[24] WESTERMANN F, HENRICH K O, WEI J S, et al. High Skp2 expression characterizes high-risk neuroblastomas independent of MYCN status[J]. Clin Cancer Res,2007,13(16):4695-4703.
[25] YANG Q, HUANG J, WU Q, et al.Acquisition of epithelial-mesenchymal transition is associated with Skp2 expression in paclitaxel-resistant breast cancer cells[J]. Br J Cancer,2014,110(8):1958-1967.
[26] DOUGLASS S, MEESON A P, OVERBECK-ZUBRZYCKA D, et al. Breast cancer metastasis:demonstration that FOXP3 regulates CXCR4 expression and the response to CXCL12[J]. J Pathol,2014,234(1):74-85.
[27] LIN H K, WANG G, CHEN Z, et al. Phosphorylation-dependent regulation of cytosolic localization and oncogenic function of Skp2 by Akt/PKB[J]. Nat Cell Biol,2009,11(4):420-432.
[28] GAO D, INUZUKA H, TSENG A, et al. Phosphorylation by Akt1 promotes cytoplasmic localization of Skp2 and impairs APCCdh1-mediated Skp2 destruction[J]. Nat Cell Biol,2009,11(4):397-408.
[29] YANG Q, CHEN C, YANG Z, et al. Suppression of breast cancer proliferation and induction of apoptosis via AKT and ERK1/2 signal transduction pathways by synthetic polypeptide derived from viral macrophage inflammatory protein Ⅱ[J]. J Huazhong Univ Sci Technolog Med Sci,2011,31(4):497-503.

[1] TIAN Hua, CHEN Yang, ZHAO Jiangang, LIU Daren, LIANG Gang, GONG Weihua, CHEN Li, WU Yulian. Effects of siRNAs targeting CD97 immune epitopes on biological behavior in breast cancer cell line MDA-MB231[J]. Journal of ZheJiang University(Medical Science), 2017, 46(4): 341-348.
[2] LI Yu, WANG Yueyue, WANG Haifeng, ZHANG Lingyu, DING Yongxing, CHEN Sulian, YANG Qingling, CHEN Changjie. Effects of lncRNA RP11-770J1.3 and TMEM25 expression on paclitaxel resistance in human breast cancer cells[J]. Journal of ZheJiang University(Medical Science), 2017, 46(4): 364-370.
[3] JIANG Yiqian, GUO Qingmin, GU Jianzhong, XU Xiaoping, AN Suhong, SU Fang, BAO Yanhong, HUANG Changxin, GUAN Xiaoxiang. Effect of microRNA-29b on proliferation and migration of breast cancer cells and its molecular mechanism[J]. Journal of ZheJiang University(Medical Science), 2017, 46(4): 349-356.
[4] CAO Peng, LENG Dongjin, LI Ying, ZHANG Ziwei, LIU Lei, LI Xiaoyan. Progress on anti-tumor molecular mechanisms of dihydroartemisinin[J]. Journal of ZheJiang University(Medical Science), 2016, 45(5): 501-507.
[5] CHEN Xiaojing, XU Junfen, YE Jing, CHENG Xiaodong, XIE Xing, LYU Weiguo. Expression of miR-let-7e-3p in cervical intraepithelial neoplasm and cervix carcinoma and its clinical significance[J]. Journal of ZheJiang University(Medical Science), 2016, 45(4): 342-348.
[6] LIN Weiren, CHEN Yatian, ZENG Linghui, YING Rongbiao, ZHU Feng. Effect of a novel EZH2 inhibitor GSK126 on prostate cancer cells[J]. Journal of ZheJiang University(Medical Science), 2016, 45(4): 356-363.
[7] LOU Pengrong, SUN Xiaonan, ZHOU Jundong, ZOU Shitao. Effect of RAD18-siRNA on proliferation and chemotherapy sensitivity of human esophageal squamous cell carcinoma ECA-109 cells[J]. Journal of ZheJiang University(Medical Science), 2016, 45(4): 364-370.
[8] WANG Cheng, WANG Wenjun, YANG Wei, YU Xiaohua, YAN Yiguo, ZHANG Jian, JIANG Zhisheng. MicroRNAs: a type of novel regulative factor for intervertebral disc degeneration[J]. Journal of ZheJiang University(Medical Science), 2016, 45(2): 170-178.
[9] WANG Xiao-jun, ZHANG Hao, ZHAN Hong-sheng, DING Dao-fang. Establishment of chondrocyte degeneration model in vitro by rat serum[J]. Journal of ZheJiang University(Medical Science), 2015, 44(3): 308-314.
[10] XIN Liao-bing, JIANG Xiu-xiu, YE Xiao-lei, WU Rui-jin, XU Kai-hong, MA Jun-yan, LIN Jun. AQP5 gene silencing inhibits proliferation and migration of ectopic endometrial glandular epithelial cells in endometriosis[J]. Journal of ZheJiang University(Medical Science), 2015, 44(3): 285-292.
[11] ZHU Hai-peng, YUN Feng, JIU Tao, et al. Inhibitory effect of inositol hexaphosphate on proliferation of LNCaP cells and its  relation to IGFBP-3 expression[J]. Journal of ZheJiang University(Medical Science), 2014, 43(5): 521-.
[12] ZHENG Yong-xia, ZHANG Cheng-wen, HUI Bin, et al. Roles of PIK3R1 gene in development of hepatocellular carcinoma[J]. Journal of ZheJiang University(Medical Science), 2014, 43(5): 559-.
[13] HAN Xiao-yu,ZHANG Lu,LIU Zhi-xian,HUANG Jing,ZHANG Meng,FANG San-hua,ZHANG Wei-ping,WEI Er-qing,LU Yun-bi . Effects of leukotriene D4 on proliferation and migration of lung epithelial A549 cells in vitro[J]. Journal of ZheJiang University(Medical Science), 2014, 43(3): 287-292.
[14] GAO Jie,WANG Rui,YANG Qingling,CHEN Changjie,WU Qiong. Effect of Oxaliplatin on cell cycle of hepatocellular carcinoma cell line HepG2[J]. Journal of ZheJiang University(Medical Science), 2013, 42(4): 437-.
[15] . Role of Wnt/β-catenin signaling in aging of mesenchymal stem cells of rats[J]. Journal of ZheJiang University(Medical Science), 2012, 41(6): 630-640.