Targeting role of CXCL10 and miR-34c-5p in invasive ductal carcinoma of breast

doi:10.3760/cma.j.cn371439-20190929-00043

Abstract

Abstract:

Objective To explore the targeted relationship between the CXC chemokine ligand 10 (CXCL10) and miR-34c-5p in invasive ductal carcinoma of breast. Methods A total of 56 cases of invasive ductal carcinoma of breast and 14 cases of benign breast tissues surgically resected and pathologically confirmed in the Third Affiliated Hospital of Sun Yat-sen University were collected from March 2016 to March 2017. The expressions of CXCL10 and miR-34c-5p in the two groups were detected using HTA2.0 and miRNA4.0 chips and verified by reverse transcription quantitative real-time PCR (RT-qPCR). The correlation between the two expressions was analyzed using Pearson correlation analysis and the targeted relationship was predicted by IPA software. The dual luciferase reporter gene system was used to verify the targets of the two and finally verified in breast cancer cell lines MCF-7 and ZR-75-30. Results The results of gene microarray showed that the mRNA expression values of CXCL10 in invasive ductal carcinoma of breast and benign breast tissues were 7.135±1.350 and 4.348±0.722, and the mRNA expression values of miR-34c-5p in the two groups were 1.309 (1.001, 2.055) and 3.948 (3.278, 4.371), with a statistically significant difference (t=7.628, P<0.001; Z=-4.405, P<0.001). Pearson correlation analysis showed that the mRNA expressions of CXCL10 and miR-34c-5p were negatively correlated (r=-0.327, P=0.004). The expression of CXCL10 mRNA was related to the expressions of human epidermal growth factor receptor-2 (t=2.415, P=0.019) and Ki-67 (t=2.483, P=0.016), and the expression of miR-34c-5p mRNA was related to the histological grading (χ ²=8.626, P=0.013) and estrogen receptor/progesterone receptor (Z=-2.195, P=0.028) in patients with invasive ductal carcinoma of breast. RT-qPCR showed that the relative expressions of CXCL10 mRNA in invasive ductal carcinoma of breast and benign breast tissues were 6.059±1.714 and 1.000±0.232, the relative expression levels of miR-34c-5p in the two groups were 0.093±0.004 and 1.000±0.244, with statistically significant differences (t=3.484, P=0.002; t=5.112, P<0.001), which was consistent with the results of microarray. The results of double luciferase reporter gene showed that, after overexpression of miR-34c-5p, the luciferase activities of WT-CXCL10 and MUT-CXCL10 were 1.117±0.040 and 1.647±0.031 in MCF-7 cells, and the luciferase activities of them were 0.885±0.051 and 1.430±0.020 in HeLa cells, with statistically significant differences (t=18.120, P<0.001; t=24.040, P<0.001). After transfection with miR-34c-5p and miR-NC respectively, the relative expression levels of CXCL10 were 0.008±0.000 and 0.012±0.000 in ZR-75-30 cells, and the relative expression levels of CXCL10 were 0.315±0.000 and 0.386±0.004 in MCF-7 cells, with statistically significant differences (t=20.384, P<0.001; t=3.473, P=0.026). Conclusion The expression of miR-34c-5p in invasive ductal carcinoma of breast is down-regulated, while the expression of CXCL10 is up-regulated. CXCL10 is the target gene of miR-34c-5p.

Key words: Breast neoplasms, CXC chemokine ligand 10, miR-34c-5p

Gao Jian, Jin Yi, Lin Jie, Lin Sheng, Duan Shan. Targeting role of CXCL10 and miR-34c-5p in invasive ductal carcinoma of breast[J]. Journal of International Oncology, 2020, 47(7): 391-396.

Figures/Tables 2

临床病理特征^a	例数	CXCL10 mRNA( $x ˉ$ ±s)	t/F值	P值	miR-34c-5p mRNA[M (P₂₅,P₇₅)]	Z/χ²值	P值
年龄(岁)
≤50	26	6.861±1.431	-1.472	0.147	1.448(1.077,2.614)	-0.378	0.706
<50	30	7.373±1.175			1.286(0.946,2.047)
组织学分级
Ⅰ	7	7.814±1.700			0.808(0.600,1.122)
Ⅱ	34	6.867±1.310	2.369	0.104	1.466(1.151,2.205)	8.626	0.013
Ⅲ	13	7.559±1.003			1.291(1.005,2.117)
ER/PR
单/双阳性	40	6.954±1.238	-1.656	0.103	1.466(1.142,2.055)	-2.195	0.028
双阴性	16	7.588±1.427			1.027(0.746,1.959)
Her-2
阳性	22	7.656±1.325	2.415	0.019	1.244(0.914,1.704)	-1.320	0.187
阴性	32	6.803±1.242			1.497(1.124,2.504)
Ki-67
≥14%	45	7.358±1.315	2.483	0.016	1.291(0.942,2.055)	-0.458	0.647
<14%	10	6.270±0.889			1.365(1.151,2.261)

References 24

[1]	Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018,68(6):394-424. DOI: 10.3322/caac.21492. doi: 10.3322/caac.21492 pmid: 30207593
[2]	中国抗癌协会乳腺癌专业委员会. 中国晚期乳腺癌临床诊疗专家共识(2018版)[J]. 中华肿瘤杂志, 2018,40(9):703-713. DOI: 10.3760/cma.j.issn.0253-3766.2018.09.013.
[3]	Karin N, Razon H. Chemokines beyond chemo-attraction: CXCL10 and its significant role in cancer and autoimmunity[J]. Cytokine, 2018,109:24-28. DOI: 10.1016/j.cyto.2018.02.012. doi: 10.1016/j.cyto.2018.02.012 pmid: 29449068
[4]	Tokunaga R, Zhang W, Naseem M, et al. CXCL9, CXCL10, CXCL11/CXCR3 axis for immune activation—a target for novel cancer therapy[J]. Cancer Treat Rev, 2018,63:40-47. DOI: 10.1016/j.ctrv.2017.11.007. doi: 10.1016/j.ctrv.2017.11.007 pmid: 29207310
[5]	Bronger H, Singer J, Windmüller C, et al. CXCL9 and CXCL10 predict survival and are regulated by cyclooxygenase inhibition in advanced serous ovarian cancer[J]. Br J Cancer, 2016,115(5):553-563. DOI: 10.1038/bjc.2016.172. pmid: 27490802
[6]	Goldberg-Bittman L, Neumark E, Sagi-Assif O, et al. The expression of the chemokine receptor CXCR3 and its ligand, CXCL10, in human breast adenocarcinoma cell lines[J]. Immunol lett, 2004,92(1-2):171-178. DOI: 10.1016/j.imlet.2003.10.020. pmid: 15081542
[7]	Liu M, Guo S, Stiles JK. The emerging role of CXCL10 in cancer (review)[J]. Oncol Lett, 2011,2(4):583-589. DOI: 10.3892/ol.2011.300. pmid: 22848232
[8]	Hermeking H. The miR-34 family in cancer and apoptosis[J]. Cell Death Differ, 2010,17(2):193-199. DOI: 10.1038/cdd.2009.56. pmid: 19461653
[9]	Zhang L, Liao Y, Tang L. MicroRNA-34 family: a potential tumor suppressor and therapeutic candidate in cancer[J]. J Exp Clin Cancer Res, 2019,38(1):53. DOI: 10.1186/s13046-019-1059-5. doi: 10.1186/s13046-019-1059-5 pmid: 30717802
[10]	Li F, Chen H, Huang Y, et al. miR-34c plays a role of tumor suppressor in HEC 1-B cells by targeting E2F3 protein[J]. Oncol Rep, 2015,33(6):3069-3074. DOI: 10.3892/or.2015.3894. doi: 10.3892/or.2015.3894 pmid: 25846116
[11]	夏丽华. 探讨女性乳腺浸润性导管癌的乳腺X线影像学特征[J]. 中外女性健康研究, 2017, (8):14, 83. DOI: 10.3969/j.issn.2096-0417.2017.08.010.
[12]	Fan L, Strasser-Weippl K, Li JJ, et al. Breast cancer in China[J]. Lancet Oncol, 2014,15(7):e279-e289. DOI: 10.1016/S1470-2045(13)70567-9. doi: 10.1016/S1470-2045(13)70567-9 pmid: 24872111
[13]	Ejaeidi AA, Craft BS, Puneky LV, et al. Hormone receptor-independent CXCL10 production is associated with the regulation of cellular factors linked to breast cancer progression and metastasis[J]. Exp Mol Pathol, 2015,99(1):163-172. DOI: 10.1016/j.yexmp.2015.06.002. pmid: 26079660
[14]	Narita D, Seclaman E, Anghel A, et al. Altered levels of plasma chemokines in breast cancer and their association with clinical and pathological characteristics[J]. Neoplasma, 2016,63(1):141-149. DOI: 10.4149/neo_2016_017. doi: 10.4149/neo_2016_017 pmid: 26639244
[15]	Jafarzadeh A, Fooladseresht H, Nemati M, et al. Higher circulating levels of chemokine CXCL10 in patients with breast cancer: evalu-ation of the influences of tumor stage and chemokine gene polymorphism[J]. Cancer Biomark, 2016,16(4):545-554. DOI: 10.3233/CBM-160596. doi: 10.3233/CBM-160596 pmid: 27002757
[16]	Bu H, Shu B, Gao F, et al. Spinal IFN-γ-induced protein-10 (CXCL10) mediates metastatic breast cancer-induced bone pain by activation of microglia in rat models[J]. Breast Cancer Res Treat, 2014,143(2):255-263. DOI: 10.1007/s10549-013-2807-4. doi: 10.1007/s10549-013-2807-4 pmid: 24337539
[17]	夏红强, 何建蓉. Ki-67、EGFR、HER-2和P53在乳腺癌中的表达及其相关性[J]. 临床肿瘤学杂志, 2011,16(2):139-143. DOI: 10.3969/j.issn.1009-0460.2011.02.010.
[18]	马士辉, 凌飞海. 乳腺癌组织中HER-2、TOPOⅡα、Ki-67的表达及临床意义[J]. 哈尔滨医药, 2016,36(5):504-506.
[19]	Wang Y, Wang X, Tang J, et al. The study of mechanism of miR-34c-5p targeting FLOT2 to regulate proliferation, migration and invasion of osteosarcoma cells[J]. Artif Cells Nanomed Biotechnol, 2019,47(1):3559-3568. DOI: 10.1080/21691401.2019.1640714. doi: 10.1080/21691401.2019.1640714 pmid: 31446795
[20]	Li F, Chen H, Huang Y, et al. miR-34c plays a role of tumor suppressor in HEC 1-B cells by targeting E2F3 protein[J]. Oncol Rep, 2015,33(6):3069-3074. DOI: 10.3892/or.2015.3894. pmid: 25846116
[21]	Córdova-Rivas S, Fraire-Soto I, Mercado-Casas Torres A, et al. 5p and 3p strands of miR-34 family members have differential effects in cell proliferation, migration, and invasion in cervical cancer cells[J]. Int J Mol Sci, 2019,20(3):545. DOI: 10.3390/ijms20030545.
[22]	Re M, Magliulo G, Gioacchini FM, et al. Expression levels and clinical significance of miR-21-5p, miR-let-7a, and miR-34c-5p in laryngeal squamous cell carcinoma[J]. Biomed Res Int, 2017,2017:3921258. DOI: 10.1155/2017/3921258. doi: 10.1155/2017/3781525 pmid: 29457024
[23]	Si W, Li Y, Shao H, et al. MiR-34a inhibits breast cancer proliferation and progression by targeting Wnt1 in Wnt/β-Catenin signaling pathway[J]. Am J Med Sci, 2016,352(2):191-199. DOI: 10.1016/j.amjms.2016.05.002. doi: 10.1016/j.amjms.2016.05.002 pmid: 27524218
[24]	Xu M, Li D, Yang C, et al. MicroRNA-34a inhibition of the TLR signaling pathway via CXCL10 suppresses breast cancer cell invasion and migration[J]. Cell Physiol Biochem, 2018,46(3):1286-1304. DOI: 10.1159/000489111. doi: 10.1159/000489111 pmid: 29689563