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Zoledronik asit uygulamasının meme kanseri hücre hattı (mcf 7) kanser kök hücrelerinin üzerine otofaji gen seviyesindeki etkilerinin araştırılması

Year 2019, Volume: 58 Issue: 1, 32 - 38, 14.03.2019

Fatih Oltulu Ayşegül Uysal Hadi Rouhrazi Kubilay Doğan Kılıç Duygu Çalık Kocatürk Gülperi Öktem

https://doi.org/10.19161/etd.417985

Abstract

Amaç: Zoledronik asit (ZA) uygulanmış meme kanser kök hücrelerinde (KKH),
kanser kök hücresi ve otofaji ile ilişkili gen ekspresyonlarındaki
farklılıkları incelemek.



Gereç ve Yöntem: CD44yüksek/CD24yüksek
yüzey belirteçleri pozitif olan MCF-7 meme kanseri hücre hattından KKH’ler akış
sitometri metodu ile izole edilmiştir. ZA tedavisi uygulanmış ve uygulanmamış
olan KKH gruplarının toplam RNA izolasyonu için cDNA sentezlenmiştir. Son
olarak 41 adet otofaji için başlıca ilişkili olabilecek genlerin ekspresyon
seviyeleri qRT-PCR analizi ile incelenmiştir.



Bulgular: ZA tedavisi uygulanan grup tedavi
uygulanmayan grup ile karşılaştırıldığında TGM2, TNF, HSPA8 genlerinin azalmış
regüle olduğu ve HSP90AA1 geninin artmış regüle olduğu görülmüştür.



Sonuç: Otofaji ile ilişkili genlerin ekspresyon profillerinde ZA tedavisi
uygulanan ve uygulanmayan gruplar arasında anlamlı farklar bulunmuştur. Bu
bulgular, KKH’lerine karşı hedef olabilecek daha etkili tedavi ajanları
geliştirilebilmesi için önemli katkılar sağlayabilir.

Keywords

Otofaji kanser kök hücresi zoledronik asit tümör nekrozis faktör

References

  • Ghoncheh M, Pournamdar Z, Salehiniya H. Incidence and mortality and epidemiology of breast cancer in the world. Asian Pac J Cancer Prev 2016;17:43-6.
  • Weigelt B, Peterse JL, van’t Veer LJ. Breast cancer metastasis: Markers and models. Nat Rev Cancer 2005;5(8):591-602.
  • Aft R. Protection of bone in premenopausal women with breast cancer: Focus on zoledronic acid. Int J Womens Heal 2012;4:569-76.
  • Barrett-Lee P, Casbard A, Abraham J, et al. Oral ibandronic acid versus intravenous zoledronic acid in treatment of bone metastases from breast cancer: A randomised, open label, non-inferiority phase 3 trial. Lancet Oncol 2014;15(1):114-22.
  • Coxon FP, Rogers MJ. The role of prenylated small GTP-binding proteins in the regulation of osteoclast function. Calcif Tissue Int 2003;72(1):80-4.
  • Body JJ, Bartl R, Burckhardt P, et al. Current use of bisphosphonates in oncology. J Clin Oncol 1998;16(2):3890-9.
  • Dedes PG, Gialeli C, Tsonis AI, et al. Expression of matrix macromolecules and functional properties of breast cancer cells are modulated by the bisphosphonate zoledronic acid. Biochim Biophys Acta 2012;1820(12):1926-39.
  • Mani J, Vallo S, Barth K, et al. Zoledronic acid influences growth, migration and invasive activity of prostate cancer cells in vitro. Prostate Cancer Prostatic Dis 2012;153(3):250-5.
  • Lin J-F, Lin Y-C, Lin Y-H, et al. Zoledronic acid induces autophagic cell death in human prostate cancer cells. J Urol 2011;185(4):1490-6.
  • Ge XY, Yang LQ, Jiang Y, Yang WW, Fu J, Li SN. Reactive oxygen species and autophagy associated apoptosis and limitation of clonogenic survival induced by zoledronic acid in salivary adenoid cystic carcinoma cell line SACC-83. PLoS One 2014;9(6).e101207.
  • Yu Z, Pestell TG, Lisanti MP, Pestell RG. Cancer stem cells. Int J Biochem Cell Biol. 2012;44(12):2144-51.
  • Singh SK, Clarke ID, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors. Cancer Res 2003;63(18):5821-8.
  • La Porta CA. Mechanism of drug sensitivity and resistance in melanoma. Curr Cancer Drug Targets 2009;9(3):391-7.
  • Yin G, Alvero a B, Craveiro V, et al. Constitutive proteasomal degradation of TWIST-1 in epithelial-ovarian cancer stem cells impacts differentiation and metastatic potential. Oncogene 2013;32(1):39-49.
  • Levine B, Yuan J. Autophagy in cell death: An innocent convict? J Clin Invest 2005;115(10):2679-88.
  • Betin VMS, Lane JD. Caspase cleavage of Atg4D stimulates GABARAP-L1 processing and triggers mitochondrial targeting and apoptosis. J Cell Sci 2009;122(14):2554-66.
  • Guan JL, Simon AK, Prescott M, et al. Autophagy in stem cells. Autophagy 2013;9(6):830-49.
  • Jaggupilli A, Elkord E. Significance of CD44 and CD24 as cancer stem cell markers: An enduring ambiguity. Clin Dev Immunol 2012;2012:708036. doi: 10.1155/2012/708036
  • Ural AU. Cancer stem cells: Current concepts and therapeutic implications. In: Turksen K (ed). Adult and Embryonic Stem Cells. Totowa, NJ: Humana Press; 2012:227-35.
  • Arslan DÖ, Korkmaz G, Gözüaçık D. Otofaji: Bir hücresel stres yanıtı ve ölüm mekanizması. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi 2011;4(2):184-94.
  • Wiley SR, Schooley K, Smolak PJ, et al. Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 1995;3(6):673-82.
  • Ashkenazi A, Pai RC, Fong S, et al. Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 1999;104(2):155-62.
  • Kelekar A. Introduction to the review series autophagy in higher eukaryotas--a matter of survival or death. Autophagy 2008;4(5):555-6.
  • Yu L. Regulation of an ATG7-beclin 1 Program of autophagic cell death by caspase-8. Science 20044;304(5676):1500-2.
  • Yu L, Wan F, Dutta S, et al. Autophagic programmed cell death by selective catalase degradation. Proc Natl Acad Sci 2006;103(13):4952-7.
  • Singh A, Ni J, Aggarwal BB. Death domain receptors and their role in cell demise. J Interferon Cytokine Res 1998;18(7):439-50.
  • Rothe M, Pan MG, Henzel WJ, Ayres TM, Goeddel D V. The TNFR2-TRAF signaling complex contains two novel proteins related to baculoviral inhibitor of apoptosis proteins. Cell 1995;83(7):1243-52.
  • Chu Z-L, McKinsey TA, Liu L, Gentry JJ, Malim MH, Ballard DW. Suppression of tumor necrosis factor-induced cell death by inhibitor of apoptosis c-IAP2 is under NF-κB control. Proc Natl Acad Sci 1997;94(19):10057-62.
  • Fisher ML, Keillor JW, Xu W, Eckert RL, Kerr C. Transglutaminase Is required for epidermal squamous cell carcinoma stem cell survival. Mol Cancer Res 2015;13(7):1083-94.
  • Pastor MD, Nogal A, Molina-Pinelo S, Carnero A, Paz-Ares L. Proteomic biomarkers in lung cancer. Clin Transl Oncol 2013;15(9):671-82.
  • Murphy ME. The HSP70 family and cancer. Carcinogenesis 2013;34(6):1181-8.
  • Maehara Y, Oki E, Abe T, et al. Overexpression of the heat shock protein HSP70 family and p53 protein and prognosis for patients with gastric cancer. Oncology 2000;58(2):144-51.
  • Shan N, Zhou W, Zhang S, Zhang Y. Identification of HSPA8 as a candidate biomarker for endometrial carcinoma by using iTRAQ-based proteomic analysis. Onco Targets Ther 2016;13(9):2169.
  • Ammirante M, Rosati A, Gentilella A, et al. The activity of hsp90 alpha promoter is regulated by NF-kappa B transcription factors. Oncogene 2008;27(8):1175-8.
  • Metz K, Ezernieks J, Sebald W, Duschl A. Interleukin-4 upregulates the heat shock protein Hsp90α and enhances transcription of a reporter gene coupled to a single heat shock element. FEBS Lett 1996;385(1-2):25-8.
  • Yufu Y, Nishimura J, Nawata H. High constitutive expression of heat shock protein 90 alpha in human acute leukemia cells. Leuk Res 1992;16(6-7):597-605.
  • Jameel A, Skilton RA, Campbell TA, Chander SK, Coombes RC, Luqmani YA. Clinical and biological significance of HSP89 alpha in human breast cancer. Int J Cancer 1992;50(3):409-15.
  • Gao J, Aksoy BA, Dogrusoz U, et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal 2013;6(269):pl1.
  • Buffart TE, Carvalho B, van Grieken NCT, et al. Losses of chromosome 5q and 14q are associated with favorable clinical outcome of patients with gastric cancer. Oncologist 2012;17(5):653-62.
  • Gallegos Ruiz MI, Floor K, Roepman P, et al. Integration of gene dosage and gene expression in non-small cell lung cancer, identification of HSP90 as potential target. PLoS One 2008;3(3):e0001722.
  • Workman P, Burrows F, Neckers L, Rosen N. Drugging the cancer chaperone HSP90: Combinatorial therapeutic exploitation of oncogene addiction and tumor stress. Ann N Y Acad Sci 2007;1113(2):202-16.
  • Eustace BK, Sakurai T, Stewart JK, et al. Functional proteomic screens reveal an essential extracellular role for hsp90α in cancer cell invasiveness. Nat Cell Biol 2004;6(6):507-14.
  • Wang X, Song X, Zhuo W, et al. The regulatory mechanism of Hsp90α secretion and its function in tumor malignancy. Proc Natl Acad Sci 2009;106(50):21288-93.

Investigation of the effects of zoledronic acid in breast cancer stem cell line (mcf7) for autophagic gene level

Year 2019, Volume: 58 Issue: 1, 32 - 38, 14.03.2019

Fatih Oltulu Ayşegül Uysal Hadi Rouhrazi Kubilay Doğan Kılıç Duygu Çalık Kocatürk Gülperi Öktem

https://doi.org/10.19161/etd.417985

Abstract

Aim: In this study we examined differences in the
expression of genes associated with autophagy and CSCs for the treatment ZA
applied via taking breast cancer stem cells as models.

Materials and Methods: CSCs with CD44high/CD24high
surface markers have been isolated from the MCF-7 breast cancer cell line
with flow cytometry methods. cDNA was synthesized for ZA treatment applied and
not applied CSC group with total RNA isolation. Then, 41 essential autophagy-related
gene expression levels were analyzed by qRT-PCR array analysis.

Results: When ZA treatment administrated group compared with
non-treatment group, downregulation of TGM2, TNF, HSPA8 and upregulation of
HSP90AA1 was detected.







Conclusion: There are differences of expression
profiles of genes related to autophagy between ZA treated and non-ZA tread
group. These findings may provide important information towards us to target
CSCs to develop more effective treatments.

Keywords

Autophagy cancer stem cell zoleodronic acid TNF heat shock protein (HSP90).

References

  • Ghoncheh M, Pournamdar Z, Salehiniya H. Incidence and mortality and epidemiology of breast cancer in the world. Asian Pac J Cancer Prev 2016;17:43-6.
  • Weigelt B, Peterse JL, van’t Veer LJ. Breast cancer metastasis: Markers and models. Nat Rev Cancer 2005;5(8):591-602.
  • Aft R. Protection of bone in premenopausal women with breast cancer: Focus on zoledronic acid. Int J Womens Heal 2012;4:569-76.
  • Barrett-Lee P, Casbard A, Abraham J, et al. Oral ibandronic acid versus intravenous zoledronic acid in treatment of bone metastases from breast cancer: A randomised, open label, non-inferiority phase 3 trial. Lancet Oncol 2014;15(1):114-22.
  • Coxon FP, Rogers MJ. The role of prenylated small GTP-binding proteins in the regulation of osteoclast function. Calcif Tissue Int 2003;72(1):80-4.
  • Body JJ, Bartl R, Burckhardt P, et al. Current use of bisphosphonates in oncology. J Clin Oncol 1998;16(2):3890-9.
  • Dedes PG, Gialeli C, Tsonis AI, et al. Expression of matrix macromolecules and functional properties of breast cancer cells are modulated by the bisphosphonate zoledronic acid. Biochim Biophys Acta 2012;1820(12):1926-39.
  • Mani J, Vallo S, Barth K, et al. Zoledronic acid influences growth, migration and invasive activity of prostate cancer cells in vitro. Prostate Cancer Prostatic Dis 2012;153(3):250-5.
  • Lin J-F, Lin Y-C, Lin Y-H, et al. Zoledronic acid induces autophagic cell death in human prostate cancer cells. J Urol 2011;185(4):1490-6.
  • Ge XY, Yang LQ, Jiang Y, Yang WW, Fu J, Li SN. Reactive oxygen species and autophagy associated apoptosis and limitation of clonogenic survival induced by zoledronic acid in salivary adenoid cystic carcinoma cell line SACC-83. PLoS One 2014;9(6).e101207.
  • Yu Z, Pestell TG, Lisanti MP, Pestell RG. Cancer stem cells. Int J Biochem Cell Biol. 2012;44(12):2144-51.
  • Singh SK, Clarke ID, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors. Cancer Res 2003;63(18):5821-8.
  • La Porta CA. Mechanism of drug sensitivity and resistance in melanoma. Curr Cancer Drug Targets 2009;9(3):391-7.
  • Yin G, Alvero a B, Craveiro V, et al. Constitutive proteasomal degradation of TWIST-1 in epithelial-ovarian cancer stem cells impacts differentiation and metastatic potential. Oncogene 2013;32(1):39-49.
  • Levine B, Yuan J. Autophagy in cell death: An innocent convict? J Clin Invest 2005;115(10):2679-88.
  • Betin VMS, Lane JD. Caspase cleavage of Atg4D stimulates GABARAP-L1 processing and triggers mitochondrial targeting and apoptosis. J Cell Sci 2009;122(14):2554-66.
  • Guan JL, Simon AK, Prescott M, et al. Autophagy in stem cells. Autophagy 2013;9(6):830-49.
  • Jaggupilli A, Elkord E. Significance of CD44 and CD24 as cancer stem cell markers: An enduring ambiguity. Clin Dev Immunol 2012;2012:708036. doi: 10.1155/2012/708036
  • Ural AU. Cancer stem cells: Current concepts and therapeutic implications. In: Turksen K (ed). Adult and Embryonic Stem Cells. Totowa, NJ: Humana Press; 2012:227-35.
  • Arslan DÖ, Korkmaz G, Gözüaçık D. Otofaji: Bir hücresel stres yanıtı ve ölüm mekanizması. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi 2011;4(2):184-94.
  • Wiley SR, Schooley K, Smolak PJ, et al. Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 1995;3(6):673-82.
  • Ashkenazi A, Pai RC, Fong S, et al. Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 1999;104(2):155-62.
  • Kelekar A. Introduction to the review series autophagy in higher eukaryotas--a matter of survival or death. Autophagy 2008;4(5):555-6.
  • Yu L. Regulation of an ATG7-beclin 1 Program of autophagic cell death by caspase-8. Science 20044;304(5676):1500-2.
  • Yu L, Wan F, Dutta S, et al. Autophagic programmed cell death by selective catalase degradation. Proc Natl Acad Sci 2006;103(13):4952-7.
  • Singh A, Ni J, Aggarwal BB. Death domain receptors and their role in cell demise. J Interferon Cytokine Res 1998;18(7):439-50.
  • Rothe M, Pan MG, Henzel WJ, Ayres TM, Goeddel D V. The TNFR2-TRAF signaling complex contains two novel proteins related to baculoviral inhibitor of apoptosis proteins. Cell 1995;83(7):1243-52.
  • Chu Z-L, McKinsey TA, Liu L, Gentry JJ, Malim MH, Ballard DW. Suppression of tumor necrosis factor-induced cell death by inhibitor of apoptosis c-IAP2 is under NF-κB control. Proc Natl Acad Sci 1997;94(19):10057-62.
  • Fisher ML, Keillor JW, Xu W, Eckert RL, Kerr C. Transglutaminase Is required for epidermal squamous cell carcinoma stem cell survival. Mol Cancer Res 2015;13(7):1083-94.
  • Pastor MD, Nogal A, Molina-Pinelo S, Carnero A, Paz-Ares L. Proteomic biomarkers in lung cancer. Clin Transl Oncol 2013;15(9):671-82.
  • Murphy ME. The HSP70 family and cancer. Carcinogenesis 2013;34(6):1181-8.
  • Maehara Y, Oki E, Abe T, et al. Overexpression of the heat shock protein HSP70 family and p53 protein and prognosis for patients with gastric cancer. Oncology 2000;58(2):144-51.
  • Shan N, Zhou W, Zhang S, Zhang Y. Identification of HSPA8 as a candidate biomarker for endometrial carcinoma by using iTRAQ-based proteomic analysis. Onco Targets Ther 2016;13(9):2169.
  • Ammirante M, Rosati A, Gentilella A, et al. The activity of hsp90 alpha promoter is regulated by NF-kappa B transcription factors. Oncogene 2008;27(8):1175-8.
  • Metz K, Ezernieks J, Sebald W, Duschl A. Interleukin-4 upregulates the heat shock protein Hsp90α and enhances transcription of a reporter gene coupled to a single heat shock element. FEBS Lett 1996;385(1-2):25-8.
  • Yufu Y, Nishimura J, Nawata H. High constitutive expression of heat shock protein 90 alpha in human acute leukemia cells. Leuk Res 1992;16(6-7):597-605.
  • Jameel A, Skilton RA, Campbell TA, Chander SK, Coombes RC, Luqmani YA. Clinical and biological significance of HSP89 alpha in human breast cancer. Int J Cancer 1992;50(3):409-15.
  • Gao J, Aksoy BA, Dogrusoz U, et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal 2013;6(269):pl1.
  • Buffart TE, Carvalho B, van Grieken NCT, et al. Losses of chromosome 5q and 14q are associated with favorable clinical outcome of patients with gastric cancer. Oncologist 2012;17(5):653-62.
  • Gallegos Ruiz MI, Floor K, Roepman P, et al. Integration of gene dosage and gene expression in non-small cell lung cancer, identification of HSP90 as potential target. PLoS One 2008;3(3):e0001722.
  • Workman P, Burrows F, Neckers L, Rosen N. Drugging the cancer chaperone HSP90: Combinatorial therapeutic exploitation of oncogene addiction and tumor stress. Ann N Y Acad Sci 2007;1113(2):202-16.
  • Eustace BK, Sakurai T, Stewart JK, et al. Functional proteomic screens reveal an essential extracellular role for hsp90α in cancer cell invasiveness. Nat Cell Biol 2004;6(6):507-14.
  • Wang X, Song X, Zhuo W, et al. The regulatory mechanism of Hsp90α secretion and its function in tumor malignancy. Proc Natl Acad Sci 2009;106(50):21288-93.
There are 43 citations in total.

Details

Primary Language Turkish
Subjects Health Care Administration
Journal Section Research Articles
Authors

Fatih Oltulu 0000-0001-6475-642X

Ayşegül Uysal 0000-0002-9919-2254

Hadi Rouhrazi 0000-0002-1367-064X

Kubilay Doğan Kılıç 0000-0002-9484-0777

Duygu Çalık Kocatürk 0000-0001-5995-8967

Gülperi Öktem 0000-0001-6227-9519

Publication Date March 14, 2019
Submission Date January 26, 2018
Published in Issue Year 2019Volume: 58 Issue: 1

Cite

Vancouver Oltulu F, Uysal A, Rouhrazi H, Kılıç KD, Çalık Kocatürk D, Öktem G. Zoledronik asit uygulamasının meme kanseri hücre hattı (mcf 7) kanser kök hücrelerinin üzerine otofaji gen seviyesindeki etkilerinin araştırılması. EJM. 2019;58(1):32-8.
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