Abstract
OBJECTIVE: Heat shock protein 90 (HSP90) is an ATPdependent
molecular chaperone required for the stability
and function of numerous oncogenic signaling proteins
that determine the hallmarks of cancer. Therefore,
HSP90 is known as a molecular therapeutic target for
the prevention and treatment of cancer. Geldanamycin
(GA) was the first identified natural Hsp90 inhibitor.
Inhibition of Hsp90 results in the induction of apoptosis
in cancer cells, and may be accompanied by a reduction
in chemotherapy resistance. This study is aimed to
investigate, on transcriptional and protein levels, the
synergic effects of unaccompanied and/or combined use
of Geldanamycin (GA) and Trichostatin A (TSA) on the
apoptotic pathway of human bladder cancer cell line T24.
MATERIAL AND METHODS: The effect of geldanamisin
(0–30 µM) on cell viability were determined by WST-1.
The variation in the expression levels of CASP3 genes was
transcriptionally determined by quantitative real-time
PCR. The translational expressional levels of caspase-3,
Bax and Bcl2 genes were performed by western blot
method.
RESULTS: The use of GA alone and GA+TSA combinati- on significantly down-regulated the antiapoptotic gene
Bcl2 while Caspase-3 and Bax expression were increased
at translational levels. Cas3 mRNA level was also increa- sed with respect to control (p<0.05).
CONCLUSIONS: The use of geldanamycin+TSA
combination reduced cell proliferation and induced
apoptosis. Therefore, it can be suggested that HSP90
inhibitors may offer a new approach to consider in the
treatment of bladder cancer
References
- Karkoulis, P.K., Stravopodis, D.J., Konstantakou, E.G., Voutsinas, G.E. (2013). Targeted inhibition of heat shock protein 90 disrupts multiple oncogenic signaling pathways, thus inducing cell cycle arrest and programmed cell death in human urinary bladder cancer cell lines. Cancer Cell International, 2013;13(1):11. Chehab, M., Caza, T., Skotnicki, K., et al. Targeting HSP90 in urothelial carcinoma. Oncotarget 2015;6(11):8454-73.Mitra, A.P., Cote, R.J. Molecular pathogenesis and diagnostics of bladder cancer. Annual Review of Pathology 2009; 4:251-85. Ischia, J., So, A.I. The role of heat shock proteins in bladder cancer. Nature Reviews Urology 2013; 10(7):386-95. Ma, L., Sato, F., Sato, R., et al. Dual targeting of heat shock proteins 90 and 70 promotes cell death and enhances the anticancer effect of chemotherapeutic agents in bladder cancer. Oncology Reports 2014;31(6):2482-92. Kiliccioglu I., Konac E., Varol N., Bilen CY. Proteasome and HDAC Inhibition Changes the Expression Levels of Bcl-2, Bcl-XL, Bim and Bik Proteins in Androgen-Independent PC-3 Cell Line. Gazi Medical Journal 2015;26:155-157.Lee, C.S., Kim, Y.J., Lee, S.A., Myung, S.C., Kim, W. Combined effect of HSP90 inhibitor geldanamycin and parthenolide via reactive oxygen species-mediated apoptotic process on epithelial ovarian cancer cells. Basic Clin Pharmacol Toxicol 2012;111(3):173-81.Mohammadi, A., Yaghoobi, M.M., Gholamhoseynian-Najar, A., Kalantari-Khandani, B., Sharifi, H., Saravani, M. HSP90 inhibitor enhances anti-proliferative and apoptotic effects of celecoxib on HT-29 colorectal cancer cells via increasing BAX/BCL-2 ratio. Cell Mol Biol 2016;62(12):62-67. Mcnamara, A.V., Barclay, M., Watson, A.J., Jenkins, J.R. HSP90 inhibitors sensitise human colon cancer cells to topoisomerase I poisons by depletion of key anti-apoptotic and cell cycle checkpoint proteins. Biochem Pharmaco, 2012;83(3):355-67.
Abstract
AMAÇ: Isı şok protein 90 (IŞP), ATP-bağımlı moleküler bir
şaperon olup kanserin temel özellikleri olarak tanımlanan
çok sayıdaki onkogenik sinyal proteinlerinin stabilitesi ve
fonksiyonu için gereklidir. Bu nedenle, IŞP90 kanserin önlenmesi
ve tedavisi için moleküler bir terapötik hedef olarak
görülür. Geldanamisin (GA) ilk doğal IŞP90 inhibitörüdür.
IŞP90 inhibisyonu, kanser hücrelerinde apoptosisin
indüklenmesine neden olur ve kemoterapi direncinde bir
azalmaya eşlik edebilir. Bu çalışmada amacımız, Geldanamisin
(GA) ve Trikostatin A (TSA)’ın tek başlarına ve/veya
kombinasyonlarının kullanımının, insan mesane kanser
hücre hattı T24’de transkripsiyonel ve protein düzeyinde
apoptotik yolak üzerindeki sinerjik etkilerini araştırmaktır.
GEREÇ VE YÖNTEM: Geldanamisin (0–30 µM)'nin hücre
canlılığı üzerine olan etkisi WST1 aracılığıyla
belirlenmiştir. Belirtilen ilaçların tek başlarına ve birlikte
kullanımlarının CASP3 geninin transkripsiyonel ifade
düzeylerindeki farklılıklar kantitatif eş zamanlı PCR
yöntemiyle belirlenmiştir. Kaspaz3, Bax ve Bcl2 genlerinin
translasyonel ifade düzeyleri ise western blot yöntemi ile
gerçekleştirilmiştir.
BULGULAR: Antiapoptotik gen Bcl2 ifadelenmesi, GA
ve GA+TSA kombinasyonu kullanımı sonrasında önemli
derecede azalmakta iken, kaspaz-3 ve Bax ifadelenmesi
kontrole göre translasyonel düzeyde artmıştır. Aynı
zamanda, Cas3 mRNA düzeyi de kontrole göre
artmıştır(p<0.05).
SONUÇ: GA+TSA kombinasyonu hücre proliferasyonunu
azaltmakta ve apoptoza indüklediği görülmüştür.
Bu nedenle, IŞP90 inhibitörlerinin mesane kanseri
tedavisinde yeni bir yaklaşım sunabileceğini
düşünmekteyiz.
Keywords
References
- Karkoulis, P.K., Stravopodis, D.J., Konstantakou, E.G., Voutsinas, G.E. (2013). Targeted inhibition of heat shock protein 90 disrupts multiple oncogenic signaling pathways, thus inducing cell cycle arrest and programmed cell death in human urinary bladder cancer cell lines. Cancer Cell International, 2013;13(1):11. Chehab, M., Caza, T., Skotnicki, K., et al. Targeting HSP90 in urothelial carcinoma. Oncotarget 2015;6(11):8454-73.Mitra, A.P., Cote, R.J. Molecular pathogenesis and diagnostics of bladder cancer. Annual Review of Pathology 2009; 4:251-85. Ischia, J., So, A.I. The role of heat shock proteins in bladder cancer. Nature Reviews Urology 2013; 10(7):386-95. Ma, L., Sato, F., Sato, R., et al. Dual targeting of heat shock proteins 90 and 70 promotes cell death and enhances the anticancer effect of chemotherapeutic agents in bladder cancer. Oncology Reports 2014;31(6):2482-92. Kiliccioglu I., Konac E., Varol N., Bilen CY. Proteasome and HDAC Inhibition Changes the Expression Levels of Bcl-2, Bcl-XL, Bim and Bik Proteins in Androgen-Independent PC-3 Cell Line. Gazi Medical Journal 2015;26:155-157.Lee, C.S., Kim, Y.J., Lee, S.A., Myung, S.C., Kim, W. Combined effect of HSP90 inhibitor geldanamycin and parthenolide via reactive oxygen species-mediated apoptotic process on epithelial ovarian cancer cells. Basic Clin Pharmacol Toxicol 2012;111(3):173-81.Mohammadi, A., Yaghoobi, M.M., Gholamhoseynian-Najar, A., Kalantari-Khandani, B., Sharifi, H., Saravani, M. HSP90 inhibitor enhances anti-proliferative and apoptotic effects of celecoxib on HT-29 colorectal cancer cells via increasing BAX/BCL-2 ratio. Cell Mol Biol 2016;62(12):62-67. Mcnamara, A.V., Barclay, M., Watson, A.J., Jenkins, J.R. HSP90 inhibitors sensitise human colon cancer cells to topoisomerase I poisons by depletion of key anti-apoptotic and cell cycle checkpoint proteins. Biochem Pharmaco, 2012;83(3):355-67.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Health Care Administration |
| Journal Section | Articles |
| Authors | |
| Publication Date | July 23, 2018 |
| Acceptance Date | June 20, 2018 |
| Published in Issue | Year 2018 Volume: 19 Issue: 3 |
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