Effect and Mechanism of The Breast Cancer Stromal Fibroblasts on Growth and Metastasis of MDA-MB-231 Cells Implanted Tumor_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 LIUChen ¹ , CAIWei ¹ ,  KANGHua ¹ , SUNHai-chen ² , LIUShuang ² , CUIYe-qing ² , ZHANGYan ¹ , HAITao ¹

1. Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing 100053, China;
2. Department of General Surgery Laboratory, Xuanwu Hospital, Capital Medical University, Beijing 100053, China;

Corresponding author：

KANGHua, Email: kanghua@xwh.ccmu.edu.cn

Keywords：

Breast cancer; Carcinoma associated fibroblasts; Stromal cell-derived factor-1; In vivo

DOI：

10.7507/1007-9424.20140070

Video：

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Objective To explore effect and mechanism of the carcinoma associated fibroblasts (CAFs) of breast cancer on growth and metastasis of breast cancer induced in nude mice by inoculation of CAFs and breast cancer cells. Methods Breast cancer cell line of MDA-MB-231 (abbreviated as MDA), CAFs, and normal breast tissue fibroblasts (NFs) of the same breast cancer patient were collected, and mixed with normal saline (NS) or SDF-1 ligand blockers of four nitrogen heterocyclic fourteen alka (AMD3100, abbreviated as AMD) for inoculation of nude mice in vivo. According to the different combination, 36 nude mice were randomly divided into 6 groups:MDA+NS group, NFs+NS group, MDA+NFs+NS group, MDA+NFs+AMD group, MDA+CAFs+AMD group, and MDA+CAFs+NS group. Forty six days after the inoculation and feeding, volume of tumor, metastasis of lymph node, lung or liver were observed. In addition, level of plasma SDF-1 was tested by using ELISA method, and expressions of SDF-1 mRNA and protein in tumor specimens were detected by using real-time PCR and Western blot method respectively. Results Except for NFs+ NS group, implanted tumor could be seen in nude mice of other 5 groups. In MDA+CAFs+NS group, the volume of tumor[(9.092±2.662) cm3], level of plasma SDF-1[(75.25±16.23) ng/L], and expression levels of SDF-1 mRNA (the median level was 14.714) and protein (the median level was 0.673). of tumor tissue were significantly greater or higher than those of the other 5 groups (P < 0.050). In addition, lymph node metastasis were found in 4 mice in MDA+CAFs+NS group, and 2 in MDA+NS group. The tumor metastasis of lung and liver was not found in all nude mice. Conclusions CAFs can promote growth and lymph node metastasis of breast cancer, whose mechanism is related with SDF-1 secreted by CAFs and SDF-1/CXC chemokine receptor 4 (CXCR4), signal pathway.

Citation： LIUChen, CAIWei, KANGHua, SUNHai-chen, LIUShuang, CUIYe-qing, ZHANGYan, HAITao. Effect and Mechanism of The Breast Cancer Stromal Fibroblasts on Growth and Metastasis of MDA-MB-231 Cells Implanted Tumor. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2014, 21(3): 289-294. doi: 10.7507/1007-9424.20140070 Copy

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14.	Chinni SR, Sivalogan S, Dong Z, et al. CXCL12/CXCR4 sign-aling activates Akt-1 and MMP-9 expression in prostate cancer cells:the role of bone microenvironment-associated CXCL12[J]. Prostate, 2006, 66(1):32-48.
15.	Retz MM, Sidhu SS, Blaveri E, et al. CXCR4 expression refl-ects tumor progression and regulates motility of bladder cancer cells[J]. Int J Cancer, 2005, 114(2):182-189.
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19.	Boimel PJ, Smirnova T, Zhou ZN, et al. Contribution of CXCL12 secretion to invasion of breast cancer cells[J]. Breast Cancer Res, 2012, 14(1):R23.
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22.	Mukherjee D, Zhao J. The role of chemokine receptor CXCR4 in breast cancer metastasis[J]. Am J Cancer Res, 2013, 3(1):46-57.
23.	Aboussekhra A. Role of cancer-associated fibroblasts in breast cancer development and prognosis[J]. Int J Dev Biol, 2011, 55(7-9):841-849.
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25.	Cabioglu N, Yazici MS, Arun B, et al. CCR7 and CXCR4 as novel biomarkers predicting axillary lymph node metastasis in T1 breast cancer[J]. Clin Cancer Res, 2005, 11(16):5686-5693.

1. Hendrayani SF, Al-Khalaf HH, Aboussekhra A. Curcumin triggers p16-dependent senescence in active breast cancer-associated fibroblasts and suppresses their paracrine procarcinogenic effects[J]. Neoplasia, 2013, 15(6):631-640.
2. Kang H, Mansel RE, Jiang WG. Genetic manipulation of stromal cell-derived factor-1 attests the pivotal role of the autocrine SDF-1-CXCR4 pathway in the aggressiveness of breast cancer cells[J]. Int J Oncol, 2005, 26(5):1429-1434.
3. Kang H, Watkins G, Douglas-Jones A, et al. The elevated level of CXCR4 is correlated with nodal metastasis of human breast cancer[J]. Breast, 2005, 14(5):360-367.
4. 张锋良, 康骅, 孙海晨.雌激素对人乳腺癌细胞系基质细胞衍化因子-1影响的观察[J].中华肿瘤防治杂志, 2011, 18(7):502-505.
5. 张晓耀, 康骅, 孙海晨, 等.乳腺癌间质成纤维细胞的分离、培养及鉴定[J].中国普外基础与临床杂志, 2011, 18(11):1160-1164.
6. Naito S, von Eschenbach AC, Giavazzi R, et al. Growth and metastasis of tumor cells isolated from a human renal cell carcinoma implanted into different organs of nude mice[J]. Cancer Res, 1986, 46(8):4109-4115.
7. Mao Y, Keller ET, Garfield DH, et al. Stromal cells in tumor microenvironment and breast cancer[J]. Cancer Metastasis Rev, 2013, 32(1-2):303-315.
8. Sadlonova A, Bowe DB, Novak Z, et al. Identification of mole-cular distinctions between normal breast-associated fibroblasts and breast cancer-associated fibroblasts[J]. Cancer Microenviron, 2009, 2(1):9-21.
9. Woo SU, Bae JW, Kim CH, et al. A significant correlation bet-ween nuclear CXCR4 expression and axillary lymph node metastasis in hormonal receptor negative breast cancer[J]. Ann Surg Oncol, 2008, 15(1):281-285.
10. Liang Z, Brooks J, Willard M, et al. CXCR4/CXCL12 axis promotes VEGF-mediated tumor angiogenesis through Akt signaling pathway[J]. Biochem Biophys Res Commun, 2007, 359(3):716-722.
11. Salvucci O, Bouchard A, Baccarelli A, et al. The role of CXCR4 receptor expression in breast cancer:a large tissue microarray study[J]. Breast Cancer Res Treat, 2006, 97(3):275-283.
12. Barbero S, Bonavia R, Bajetto A, et al. Stromal cell-derived factor 1alpha stimulates human glioblastoma cell growth through the activation of both extracellular signal-regulated kinases 1/2 and Akt[J]. Cancer Res, 2003, 63(8):1969-1974.
13. Yoon Y, Liang Z, Zhang X, et al. CXC chemokine receptor-4 antagonist blocks both growth of primary tumor and metastasis of head and neck cancer in xenograft mouse models[J]. Cancer Res, 2007, 67(15):7518-7524.
14. Chinni SR, Sivalogan S, Dong Z, et al. CXCL12/CXCR4 sign-aling activates Akt-1 and MMP-9 expression in prostate cancer cells:the role of bone microenvironment-associated CXCL12[J]. Prostate, 2006, 66(1):32-48.
15. Retz MM, Sidhu SS, Blaveri E, et al. CXCR4 expression refl-ects tumor progression and regulates motility of bladder cancer cells[J]. Int J Cancer, 2005, 114(2):182-189.
16. Pan J, Mestas J, Burdick MD, et al. Stromal derived factor-1(SDF-1/CXCL12) and CXCR4 in renal cell carcinoma metastasis[J]. Mol Cancer, 2006, 5:56.
17. Kato M, Kitayama J, Kazama S, et al. Expression pattern of CXC chemokine receptor-4 is correlated with lymph node metas-tasis in human invasive ductal carcinoma[J]. Breast Cancer Res, 2003, 5(5):R144-R150.
18. Elkabets M, Gifford AM, Scheel C, et al. Human tumors insti-gate granulin-expressing hematopoietic cells that promote malig-nancy by activating stromal fibroblasts in mice[J]. J Clin Invest, 2011, 121(2):784-799.
19. Boimel PJ, Smirnova T, Zhou ZN, et al. Contribution of CXCL12 secretion to invasion of breast cancer cells[J]. Breast Cancer Res, 2012, 14(1):R23.
20. Emens LA, Silverstein SC, Khleif S, et al. Toward integrative cancer immunotherapy:targeting the tumor microenvironment[J]. J Transl Med, 2012, 10:70.
21. Kang H, Watkins G, Parr C, et al. Stromal cell derived factor-1:its influence on invasiveness and migration of breast cancer cells in vitro, and its association with prognosis and survival in human breast cancer[J]. Breast Cancer Res, 2005, 7(4):R402-R410.
22. Mukherjee D, Zhao J. The role of chemokine receptor CXCR4 in breast cancer metastasis[J]. Am J Cancer Res, 2013, 3(1):46-57.
23. Aboussekhra A. Role of cancer-associated fibroblasts in breast cancer development and prognosis[J]. Int J Dev Biol, 2011, 55(7-9):841-849.
24. Xouri G, Christian S. Origin and function of tumor stroma fibroblasts[J]. Semin Cell Dev Biol, 2010, 21(1):40-46.
25. Cabioglu N, Yazici MS, Arun B, et al. CCR7 and CXCR4 as novel biomarkers predicting axillary lymph node metastasis in T1 breast cancer[J]. Clin Cancer Res, 2005, 11(16):5686-5693.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect and Mechanism of The Breast Cancer Stromal Fibroblasts on Growth and Metastasis of MDA-MB-231 Cells Implanted Tumor

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