SCD  Vol.1 No.2 , July 2011
Establishment and molecular characterization of breast cancer mesenchymal stem cell line derived from human non-metastasis breast cancer tumor
ABSTRACT
Breast cancer remains a leading cause of morbidity and mortality in women mainly because of the propensity of primary breast tumors to metastasize. It is composed of heterogeneous cell populations with different biological properties. Breast cancer-initiating cells have been recently identified in breast carcinoma as CD44+/CD24-/low cells, which display stem cell like properties. In the present study, we have isolated breast cancer stem cells from non-metastasis tumor tissue, which is presently at passage 18 and designated as human Breast Cancer Mesenchymal Stem Cells (hBCMSCs) line. These cells showed spindle shaped morphology and formed mammos-pheres as well as pluripotency clones indicating their stem cell nature. Molecular marker study confirmed mesenchymal nature as well as pluripotency, plasticity and oncogenicity of these cells. The hBCMSCs cell line may likely contain a heterogeneous population of malignant cells. Interestingly, we also found that these cells exhibit BRCA 2 mutation, which was found in Indian population. Overall, this study revealed that hBCMSCs cell line may represent a suitable in vitro model to study the mechanism of breast cancer which further leads to an identification of molecular targets for future breast cancer targeted therapy.

Cite this paper
nullPotdar, P. and Chaugule, S. (2011) Establishment and molecular characterization of breast cancer mesenchymal stem cell line derived from human non-metastasis breast cancer tumor. Stem Cell Discovery, 1, 21-28. doi: 10.4236/scd.2011.12003.
References
[1]   Smith, G.H. and Chepko, G. (2001) Mammary epithelial stem cells. Microscopy Research and Technique, 15, 190-203.

[2]   Pardal, R., Clarke, M.F. and Morrison, S.J. (2003) Applying the principles of stem-cell biology to cancer. Nature Reviews Cancer, 3, 895-902.

[3]   Lapidot, T., Sirard, C., Vormoor, J., Murdoch, B., Hoang, T., Caceres-Cortes, J., Minden, M., Paterson, B., Caligiuri, M.A. and Dick, J.E. (1994) A cell initiating human acute myeloid leukemia after transplantation into SCID mice. Nature, 367, 645-648.

[4]   Fidler, I.J. and Kripke, M.L. (1997) Metastasis results from preexisting variant cells within a malignant tumor. Science, 197, 893-895.

[5]   Al-Hajj, M., Wicha, M.S., Benito-Hernandez, A., Morrison, S.J. and Clarke, M.F. (2003) Prospective identification of tumorigenic breast cancer cells. Proceedings of the National Academy of Sciences, 100, 3983-3988.

[6]   Bonnet, D. and Dick, J.E. (1997) Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nature Medicine, 3, 730-737.

[7]   Collins, A.T., Berry, P.A., Hyde, C., Stower, M.J. and Maitland, N.J. (2005) Prospective Identification of tumorigenic prostate cancer stem cells. Cancer Research, 65, 10946-10951.

[8]   Hemmati, H.D., Nakano, I., Lazareff, J.A., Masterman-Smith, M., Geschwind, D.H., Bronner-Fraser M. and Komblurn, H.I. (2003) Cancerous stem cells can arise from pediatric brain tumors. Proceedings of the National Academy of Sciences, 100, 15178-15183.

[9]   Kim, C.F., Jackson, E.L., Woolfenden, A.E., Lawrence, S., Babar, I., Vogel, S., Crowley, D., Bronson, R.T. and Jacks, T. (2005) Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell, 121, 823-835.

[10]   O’Brien, C.A., Pollett, A., Gallinger, S. and Dick, J.E. (2007) A human colon cancer cell capable of initiating tumor growth in immunodeficient mice. Nature, 445, 106-110.

[11]   Potdar, P.D. and Subedi, R.P. (2011) Defining Molecular Phenotypes of Mesenchymal and hematopoietic Stem Cells derived from Peripheral blood of Acute Lymphocytic Leukemia patients for regenerative stem cell therapy. Journal of Stem cells & Regenerative Medicine, 7, 29-40.

[12]   Grimshaw, M.J., Cooper, L., Papazisis, K., Coleman, J.A., Bohnenkamp, H.R., Chiapero-Stanke, L., Taylor-Papa- dimitriou, J. and Burchell, J.M. (2008) Mammosphereculture of metastatic breast cancer cells enriches for tumorigenic breast cancer cells. Breast Cancer Research, 10, 52.

[13]   Wicha, M., Liu, S. and Dontu, G. (2006) Cancer stem cells: an old idea – a paradigm shift. Cancer Research, 66, 1883-1890.

[14]   Barry, F.P., Boynton, R.E., Haynesworth, S., Murphy, J.M. and Zaia, J. (1999) The monoclonal antibody SH-2, raised against human mesenchymal stem cells, recognizes an epitope on endoglin (CD105). Biochemical and Biophysical Research Communications, 265, 134-139.

[15]   Healy, L., May, G., Gale, K., Grosvel, F., Greaves, M. and Enver, T. (1995) The stem cell antigen CD34 functions as a regulator of hemopoietic cell adhesion. Proceedings of the National Academy of Sciences, 92, 12240-12244.

[16]   Ogata, K., Satoh, C., Tachibana, M., Hyodo, H., Tamura, H., Dan, K., Kimura, T., Sonoda, Y. and Tsuji, T. (2005) Identification and hematopoietic potential of CD45-clonal cells with very immature phenotypes (CD45- Cd34-Cd38-Lin-) in patients with myelodysplastic syndromes. Stem Cell, 23, 619-630.

[17]   Olamura-Nakanishi, S., Saito, M., Niwa, H. and Isshilcawa, F. (2005) Oct-3/4 and Sox2 regulate Oct-3/4 gene in embryonic stem cells. Journal of Biological Chemistry, 22, 5307-5317.

[18]   Prescott, S.M. and Fitzpatrick, F.A. (2000) Cyclooxygenase-2 and carcinogenesis. Biochimica et Biophysica Acta, 1470, M69-M78.

[19]   Katzenellenbogen, R.A., Baylin, S.B. and Herman, J.G. (1999) Hypermethylation of the DAP-Kinase CpG island is a common alteration in B-cell malignancies. Blood, 93, 4347-4353.

[20]   William, R.L., Hilton, D.J., Pease, S., Willson, T.A., Stewart, C.L., Gearing, D.P., Wagner, E.F., Metcalf, D., Nicola, N.A. and Gough, N.M. (1988) Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature, 336, 684-687.

[21]   Draffin, J.E., McFarlane, S., Hill, A., Johnston, P.G. and Waugh, D.J. (2004) CD44 potentiates the adherence of metastatic prostate and breast cancer cells to bone marrow endothelial cells. Cancer Research, 64, 5702-5711.

[22]   Schabeth, H., Runz, S., Joumaa, S. and Altevogt, P. (2006) CD24 affects CXCR4 function in pre-β lymphocytes and breast carcinoma cells. Journal of Cell Science, 119, 314-325.

[23]   Potdar, P.D. and Bisht, S. L. (2009) Identification and screening for novel mutations in exon 10 and exon 11 of BRCA 1 and BRCA 2 genes in hereditary and sporadic breast cancer patients in Indian population- concept for biomarkers for early detection. Annals of Oncology, 20, 51.

[24]   Potdar, P.D. and Sutar, J. P. (2010) Establishment and molecular characterization of mesenchymal stem cell lines derived from human visceral & subcutaneous adipose tissues. Journal of Stem cells & Regenerative Medicine, 6, 1-10.

[25]   Knupfer, H. and Preiss, R. (2007) Significance of interleukin-6 (IL-6) in breast cancer. Breast Cancer Res Treat, 102, 129-35.

[26]   Shi, W., Wang, H., PanYijie, G., Yunqian, G., Pei, G., (2006) Regulation of the Pluripotency Marker Rex-1 by NANOG and SOX2. Journal of Biological Chemistry, 281, 23319-23325.

[27]   Ku, N., et al. (1997) Mutation of human keratin 18 in association with cryptogenic cirrhosis. Journal of Clinical Investigation, 99, 19-23.

[28]   Malumbres, M. and Barbacid, M. (2001) To cycle or not to cycle: a critical decision in cancer. Nature Reviews Cancer, 1, 222-231.

[29]   Nesbit, C.E., Tersak, J.M. and Prochownik, E.V. (1999) MYC oncogenes and human neoplastic disease. Oncogene, 18, 3004-3016.

[30]   Bhargava, R., Gerald, W.L., Li, A.R., Pan, Q., Lal, P., Ladanyi, M. and Chen, B. (2005) EGFR gene amplification in breast cancer: Correlation with epidermal growth factor receptor mRNA and protein expression and HER-2 status and absence of EGFR-activating mutations. Modern Pathology, 18, 1027-1033.

[31]   Swellam, M., Ismail, M., Eissa, S., Hamdy, M. and Mokhtar, N. (2004) Emerging role of P53, Bcl-2 and telomerase activity in egyptian breast cancer patients. IUBMB Life, 56, 483-490.

[32]   Chen, Y., Shi, L., Zhang, L., Li, R., Liang, J., Yu, W., Sun, L., Yang, X., Wang, Y., Zhang, Y. and Shang, Y. (2008) The molecular mechanism governing the oncogenic potential of SOX2 in breast cancer. The Journal of Biological Chemistry, 283, 17969-17978.

[33]   Charafe-Jauffret, E., et al., (2006) Gene expression profiling of breast cell lines identifies potential new basal markers. Oncogene, 25, 2273-2284.

[34]   Smith, M.C.P., Luker, K.E., Garbow, J.R., et al., (2004) CXCR4 regulates growth of both primary and metastatic breast cancer. Cancer Research, 64, 8604-8612.

 
 
Top