DOI QR코드

DOI QR Code

Differentially Expressed Proteins in ER+ MCF7 and ER- MDA-MB-231 Human Breast Cancer Cells by RhoGDI-α Silencing and Overexpression

  • Hooshmand, Somayeh (Department of Obstetrics and Gynaecology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia) ;
  • Ghaderi, Abbas (Cancer Proteomics and Biomarkers Lab, Shiraz Institute for Cancer Research, University of Medical Sciences) ;
  • Yusoff, Khatijah (Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia) ;
  • Thilakavathy, Karuppiah (Department of Obstetrics and Gynaecology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia) ;
  • Rosli, Rozita (Department of Obstetrics and Gynaecology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia) ;
  • Mojtahedi, Zahra (Cancer Proteomics and Biomarkers Lab, Shiraz Institute for Cancer Research, University of Medical Sciences)
  • Published : 2014.04.01

Abstract

Background: The consequence of Rho GDP dissociation inhibitor alpha (RhoGDI${\alpha}$) activity on migration and invasion of estrogen receptor positive ($ER^+$) and negative ($ER^-$) breast cancer cells has not been studied using the proteomic approach. Changes in expression of RhoGDI${\alpha}$ and other proteins interacting directly or indirectly with RhoGDI${\alpha}$ in MCF7 and MDA-MB-231, with different metastatic potentials is of particular interest. Materials and Methods: $ER^+$ MCF7 and ER- MDA-MB-231 cell lines were subjected to two-dimensional electrophoresis (2-DE) and spots of interest were identified by matrix-assisted laser desorption/ionization time of- flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry (MS) analysis after downregulation of RhoGDI${\alpha}$ using short interfering RNA (siRNA) and upregulated using GFP-tagged ORF clone of RhoGDI${\alpha}$. Results: The results showed a total of 35 proteins that were either up- or down-regulated in these cells. Here we identifed 9 and 15 proteins differentially expressed with silencing of RhoGDI${\alpha}$ in MCF-7 and the MDA-MB-231 cells, respectively. In addition, 10 proteins were differentially expressed in the upregulation of RhoGDI${\alpha}$ in MCF7, while only one protein was identified in the upregulation of RhoGDI${\alpha}$ in MDA-MB-231. Based on the biological functions of these proteins, the results revealed that proteins involved in cell migration are more strongly altered with RhoGDI-${\alpha}$ activity. Although several of these proteins have been previously indicated in tumorigenesis and invasiveness of breast cancer cells, some ohave not been previously reported to be involved in breast cancer migration. Hence, these proteins may serve as useful candidate biomarkers for tumorigenesis and invasiveness of breast cancer cells. Conclusions: Future studies are needed to determine the mechanisms by which these proteins regulate cell migration. The combination of RhoGDI${\alpha}$ with other potential biomarkers may be a more promising approach in the inhibition of breast cancer cell migration.

Keywords

Proteomics;biomarkers;RhoGDI${\alpha}$;$ER^+$ MCF7;$ERT^-$ MDA-MB-231;breast cancer cells

References

  1. Zhang SJ, Hu Y, Qian HL, et al (2013). Expression and significance of ER, PR, VEGF, CA15-3, CA125 and CEA in judging the prognosis of breast cancer.. Asian Pac J Cancer Prev, 14, 3937-40. https://doi.org/10.7314/APJCP.2013.14.6.3937
  2. Suhane S, Berel D, Ramanujan VK (2011). Biomarker signatures of mitochondrial NDUFS3 in invasive breast carcinoma. Biochem Biophys Res Commun, 412, 590-95. https://doi.org/10.1016/j.bbrc.2011.08.003
  3. Srisomsap C, Sawangareetrakul P, Subhasitanont P, et al (2004). Proteomic analysis of cholangiocarcinoma cell line. Proteomics, 4, 1135-44. https://doi.org/10.1002/pmic.200300651
  4. Strande V, Canelle L, Tastet C, et al (2009). The proteome of the human breast cancer cell line MDA-MB-231: Analysis by LTQ-Orbitrap mass spectrometry. Proteomics Clin, 3, 41-50. https://doi.org/10.1002/prca.200800083
  5. Stresing V, Baltziskueta E, Rubio N, et al (2013). Peroxiredoxin 2 specifically regulates the oxidative and metabolic stress response of human metastatic breast cancer cells in lungs. Oncogene, 32, 724-35. https://doi.org/10.1038/onc.2012.93
  6. Tari AM, Hung MC, Li K, Lopez-Berestein G (1999). Growth inhibition of breast cancer cells by Grb2 downregulation is correlated with inactivation of mitogen-activated protein kinase in EGFR, but not in ErbB2, cells. Oncogene, 18, 1325-32. https://doi.org/10.1038/sj.onc.1202422
  7. Undyala VV, Dembo M, Cembrola K, et al (2008). The calpain small subunit regulates cell-substrate mechanical interactions during fibroblast migration. J Cell Sci, 121, 3581-8. https://doi.org/10.1242/jcs.036152
  8. Wilkins MR, Pasquali C, Appel RD, et al (1996). From proteins to proteomes: large scale protein identification by two-dimensional electrophoresis and amino acid analysis. Biotechnology, 14, 61-5. https://doi.org/10.1038/nbt0196-61
  9. Yamada Y, Arao T, Gotoda T, et al (2008). Identification of prognostic biomarkers in gastric cancer using endoscopic biopsy samples. Cancer Sci, 99, 2193-9. https://doi.org/10.1111/j.1349-7006.2008.00935.x
  10. Yan Y, Mahotka C, Heikaus S, et al (2004). Disturbed balance of expression between XIAP and Smac/DIABLO during tumour progression in renal cell carcinomas. Br J Cancer, 91, 1349-57. https://doi.org/10.1038/sj.bjc.6602127
  11. Rho SB, Song YJ, Lim MC, et al (2012). Programmed cell death 6 (PDCD6) inhibits angiogenesis through PI3K/mTOR/p70S6K pathway by interacting of VEGFR-2. Cell Signal, 24, 131-9. https://doi.org/10.1016/j.cellsig.2011.08.013
  12. Pluta P, Cebula-Obrzut B, Ehemann V, et al (2011). Correlation of Smac/DIABLO protein expression with the clinico-pathological features of breast cancer patients. Neoplasma, 58, 430-5. https://doi.org/10.4149/neo_2011_05_430
  13. Pola C (2012). Cancer: Antitumor duality of ApoE. Nature Medicine, 18, 1752.
  14. Reis-Filho JS, Lakhani SR (2003). The diagnosis and management of pre-invasive breast disease: genetic alterations in pre-invasive lesions. Breast Cancer Res, 5, 313-9. https://doi.org/10.1186/bcr650
  15. Ryu J, Song J, Heo J, et al (2011). Cross-regulation between protein L-isoaspartyl O-methyltransferase and ERK in epithelial mesenchymal transition of MDA-MB-231 cells. Acta Pharmacol Sin, 32, 1165-72. https://doi.org/10.1038/aps.2011.94
  16. Salicioni AM, Xi M, Vanderveer LA, et al (2000). Identification and structural analysis of human RBM8A and RBM8B: two highly conserved RNA-binding motif proteins that interact with OVCA1, a candidate tumor suppressor. Genomics, 69, 54-62. https://doi.org/10.1006/geno.2000.6315
  17. Sarvaiya HA, Yoon JH, Lazar IM (2006). Proteome profile of the MCF7 cancer cell line: a mass spectrometric evaluation. Rapid Commun Mass Spectrom, 20, 3039-55. https://doi.org/10.1002/rcm.2677
  18. Shida A, Fujioka S, Takahashi N, et al (2013). Reduced expression of Rho GDP dissociation inhibitor 2 mRNA is associated with lymph node metastasis in gastric carcinoma. Oncol Lett, 6, 463-7. https://doi.org/10.3892/ol.2013.1379
  19. Sofi GN, Sofi JN, Nadeem R, et al (2012). Estrogen receptor and progesterone receptor status in breast cancer in relation to age, histological grade, size of lesion and lymph node involvement. Asian Pac J Cancer Prev, 13, 5047-52. https://doi.org/10.7314/APJCP.2012.13.10.5047
  20. Libertini SJ, Robinson BS, Dhillon NK, et al (2005). Cyclin E both regulates and is regulated by calpain 2, a protease associated with metastatic breast cancer phenotype. Cancer Res, 65, 10700-8. https://doi.org/10.1158/0008-5472.CAN-05-1666
  21. Kanaujiya JK, Lochab S, Kapoor I, et al (2013). Proteomic identification of Profilin1 as a corepressor of estrogen receptor alpha in MCF7 breast cancer cells. Proteomics, 13, 2100-12. https://doi.org/10.1002/pmic.201200534
  22. Lai TC, Chou HC, Chen YW, et al (2010). Secretomic and proteomic analysis of potential breast cancer markers by two-dimensional differential gel electrophoresis. J Proteome Res, 9, 1302-22. https://doi.org/10.1021/pr900825t
  23. Lee S, Terry D, Hurst DR, Welch DR, Sang QX (2011). Protein signatures in human MDA-MB-231 breast cancer cells indicating a more invasive phenotype following knockdown of human endometase/matrilysin-2 by siRNA. J Cancer, 2, 165-76. https://doi.org/10.7150/jca.2.165
  24. Lin M, van Golen KL (2004). Rho-regulatory proteins in breast cancer cell motility and invasion. Breast Cancer Res Treat, 84, 49-60. https://doi.org/10.1023/B:BREA.0000018424.43445.f3
  25. Mommers EC, van Diest PJ, Leonhart AM, Meijer CJ, Baak JP (1999). Balance of cell proliferation and apoptosis in breast carcinogenesis. Breast Cancer Res Treat, 58, 163-9. https://doi.org/10.1023/A:1006396103777
  26. Nagaraja GM, Othman M, Fox BP, et al (2006). Gene expression signatures and biomarkers of noninvasive and invasive breast cancer cells: comprehensive profiles by representational difference analysis, microarrays and proteomics. Oncogene, 25, 2328-38. https://doi.org/10.1038/sj.onc.1209265
  27. Niemi M, Kervinen K, Kiviniemi H, et al (2000). Apolipoprotein E phenotype, cholesterol and breast and prostate cancer. J Epidemiol Community Health, 54, 938-9. https://doi.org/10.1136/jech.54.12.938
  28. Pan J, Sun LC, Tao YF, et al (2011). ATP synthase ecto-alpha-subunit: a novel therapeutic target for breast cancer. J Transl Med, 9, 211. https://doi.org/10.1186/1479-5876-9-211
  29. Grant AG, Flomen RM, Tizard ML, Grant DA (1992). Differential screening of a human pancreatic adenocarcinoma lambda gt11 expression library has identified increased transcription of elongation factor EF-1 alpha in tumour cells. Int J Cancer, 50, 740-5. https://doi.org/10.1002/ijc.2910500513
  30. Ding Z, Joy M, Bhargava R, et al (2013). Profilin-1 downregulation has contrasting effects on early vs late steps of breast cancer metastasis. Oncogene.???
  31. Gast MC, Schellens JH, Beijnen JH (2009). Clinical proteomics in breast cancer: a review. Breast Cancer Res Treat, 116, 17-29. https://doi.org/10.1007/s10549-008-0263-3
  32. Graff JR, Konicek BW, Vincent TM, et al (2007). Therapeutic suppression of translation initiation factor eIF4E expression reduces tumor growth without toxicity. J Clin Invest, 117, 2638-48. https://doi.org/10.1172/JCI32044
  33. Hoj BR, la Cour JM, Mollerup J, Berchtold MW (2009). ALG-2 knockdown in HeLa cells results in G2/M cell cycle phase accumulation and cell death. Biochem Biophys Res Commun, 378, 145-8. https://doi.org/10.1016/j.bbrc.2008.11.021
  34. Hooshmand S, Ghaderi A, Yusoff K, et al (2013). Downregulation of $RhoGDI\alpha$ increased migration and invasion of ER (+) MCF7 and ER (-) MDA-MB-231 breast cancer cells. Cell Adh Migr, 7, 297-303. https://doi.org/10.4161/cam.24204
  35. Hu LD, Zou HF, Zhan SX, Cao KM (2007). Biphasic expression of RhoGDI2 in the progression of breast cancer and its negative relation with lymph node metastasis. Oncol Rep, 17, 1383-9.
  36. Ji Y, Olson J, Zhang J, et al (2008). Breast cancer risk reduction and membrane-bound catechol O-methyltransferase genetic polymorphisms. Cancer Res, 68, 5997-6005. https://doi.org/10.1158/0008-5472.CAN-08-0043
  37. Bae YH, Ding Z, Zou L, et al (2009). Loss of profilin-1 expression enhances breast cancer cell motility by Ena/VASP proteins. J Cell Physiol, 219, 354-64. https://doi.org/10.1002/jcp.21677
  38. Barone I, Brusco L, Gu G, et al (2011). Loss of Rho GDIalpha and resistance to tamoxifen via effects on estrogen receptor alpha. Natl Cancer Inst, 103, 538-52. https://doi.org/10.1093/jnci/djr058
  39. Brenton JD, Carey LA, Ahmed AA, Caldas C (2005). Molecular classification and molecular forecasting of breast cancer: ready for clinical application? Clin Oncol, 23, 7350-60. https://doi.org/10.1200/JCO.2005.03.3845
  40. Carragher NO, Fonseca BD, Frame MC (2004). Calpain activity is generally elevated during transformation but has oncogene-specific biological functions. Neoplasia, 6, 53-73. https://doi.org/10.1016/S1476-5586(04)80053-8
  41. Chua PJ, Lee EH, Yu Y, et al (2010). Silencing the Peroxiredoxin III gene inhibits cell proliferation in breast cancer. Int J Oncol, 36, 359-64.
  42. Dawling S, Roodi N, Mernaugh RL, Wang X, Parl FF (2001). Catechol-O-methyltransferase (COMT)-mediated metabolism of catechol estrogens: comparison of wild-type and variant COMT isoforms. Cancer Res, 61, 6716-22.

Cited by

  1. Protein Profiles Associated with Anoikis Resistance of Metastatic MDA-MB-231 Breast Cancer Cells vol.17, pp.2, 2016, https://doi.org/10.7314/APJCP.2016.17.2.581
  2. Fatty acid synthase regulates the chemosensitivity of breast cancer cells to cisplatin-induced apoptosis vol.22, pp.6, 2017, https://doi.org/10.1007/s10495-017-1366-2
  3. Sphingomyelin synthase 2 promotes an aggressive breast cancer phenotype by disrupting the homoeostasis of ceramide and sphingomyelin vol.10, pp.3, 2019, https://doi.org/10.1038/s41419-019-1303-0