Bendamustin ile indüklenen HL60 hücre serisinde apoptozun ve hücre döngüsünde görevli genlerin ekspresyon değişimlerinin belirlenmesi
Abstract
Amaç: Akut miyeloid lösemi
(AML), hematopoetik öncül hücrelerin heterojen klonal bir hastalığıdır. Akut
promiyelositer lösemi (APL), bir AML alt tipidir ve morfolojik olarak anormal
promiyelositler ile karakterize edilir. APL’de görülen en belirgin kromozom
düzensizliği ise 15. kromozomdaki promyelositik lösemi (PML) geni ile 17.
kromozomdaki retinoik asit reseptör (RAR) alfa geninin translokasyonu sonucu
bir füzyon proteini oluşturmasıdır. Bendamustin suda çözünebilen, beyaz,
nitrojen hardal grubu ve bütirik asit yan zinciri bulundurması nedeniyle
amfoterik özellik özellik gösteren mikrokristallin tozudur. Alkilleyici ajan
olan bendamustin aynı zamanda pürin analoğu özelliği gösterir ve diğer
alkilleyici ajanlara göre toksisite profili daha ılımlıdır. Bu çalışmada,
bendamustinin APL hücre serisi olan HL60 hücreleri üzerindeki sitotoksik,
sitostatik ve apoptotik etkileri incelenmiş ve hücre döngüsü, mitotik katastrof
ve apoptoz ile ilişkili genlerinin ilaç ile muamele sonrasındaki ekspresyon
değişimleri belirlenmiştir.
Gereç ve Yöntem: Çalışmamızda
bendamustinin APL’ye model oluşturan HL60 hücreleri üzerindeki sitotoksik
etkisini WST-1 testi ile, apoptotik etkisini Annexin V ve Apo Direct Tunnel
methoduyla, sitostatik etkisini ise Cycletest Plus DNA Reagent Kit ile
değerlendirilmiştir. Ayrıca bendamustinin HL60 hüce serilerindeki hücre döngüsü,
mitotik katastrof ve apoptoz ile ilişkili genlerinin mRNA ekspresyonlarına
etkisi kantitatif RT-PCR ile rölatif olarak belirlenmiştir.
Bulgular: Bendamustinin
sitotoksik etkisinin HL60 hücre serisi üzerinde yüksek dozlarda etkili olduğu
belirlenmiştir. Bununla birlikte, apoptotik etkisinin her iki yöntem ile de
doza bağımlı olarak anlamlı bir şekilde arttığı belirlenmiştir. Bendamustin ile
48 saat muamele sonrasında HL-60 hücrelerinde apoptoz ile ilişkili olduğu
bilinen CASP3, CASP8, CASP9 ekspresyonlarını sırasıyla 2.4, 5.7, 6.8 –
kat arttırdığı saptanmıştır. Ayrıca mitotik katastrofi ile ilişkili olduğu
bilinen Plk1 ekspresyonun 2.5-kat azaldığını ve tümör supresör olduğu bilinen
Plk2 ekspresyonunda ise 17 katlık bir artma belirlenmiştir.
Sonuç: Bulgularımız, bendamustinin apoptoz ve mitotik katastrof
gibi hücre ölüm mekanizmaları üzerinde etkili olduğunu ve APL’ nin tamamlayıcı
tedavisinde alternatif bir tedavi olarak kullanılabileceğini göstermektedir.
Keywords
References
- 1. Ozegowski W, Krebs D. w-[bis-(chlorethyl)-amino-benzimidazolyl-(2)]-propionic or butyric acids as potential cytostatic agents J Prakt Chem 1963; 20:178–86.2. Hartmann M, Zimmer CH. Investigation of cross-link formation in DNA by the alkylating cytostatica IMET 3106, 3393 and 3943 Biochim Biophys Acta 1972; 287:386–9.3. Strumberg D, Harstrick A, Doll K et al. Bendamustine hydrochloride activity against doxorubicine-resistant human breast cancer cell lines Anticancer Drugs 1996; 7:415–21.4. Staib P, Schinkothe T, Dimski T et al. In vitro modulation of ara-CTP accumulation in fresh AML cells by bendamustine in comparison with fludarabine, 2-CDA and gemcitabine Blood 1999; 94:63.5. Hiraoka N, Kikuchi J, Yamauchi T, Koyama D, Wada T, Uesawa M, Akutsu M, Mori S, Nakamura Y, Ueda T, Kano Y. Furukawa Purine analog-like properties of bendamustine underlie rapid activation of DNA damage response and synergistic effects with pyrimidine analogues in lymphoid malignancies Y.PLoS One 2014; 13;9(3):e90675. doi: 10.1371/journal.pone.0090675.6. Kost SE, Bouchard ED, LaBossière É, Ye X, Queau ML, Liang WS, Banerji V, Gibson SB, Katyal S,Johnston JB. Cross-resistance and synergy with bendamustine in chronic lymphocytic leukemia Leuk Res. 2016 ;50:63-71. doi: 10.1016/j.leukres.2016.09.016.7. Scutaru AM, Wenzel M, Scheffler H, Wolber G, Gust R. Optimization of the N-lost drugs melphalan and bendamustine: synthesis and cytotoxicity of a new set of dendrimer-drug conjugates as tumor therapeutic agents. Bioconjug Chem. 2010; 21(10):1728-43. doi: 10.1021/bc900453f.8. Inoue M, Honma Y, Urano T, Suzumiya J.Japanese apricot extract (MK615) potentiates bendamustine-induced apoptosis via impairment of the DNA damage response in lymphoma cells. Oncol Lett. 2017 Jul;14(1):792-800. doi: 10.3892/ol.2017.6219.9. Palombi M, Niscola P, Tendas A, Trawinska MM, Scaramucci L, Giovannini M, Fratoni S, Perrotti A, de Fabritiis P. Simultaneous occurrence of large B-cell non-Hodgkin lymphoma and acute myeloid leukaemia in an elderly patient: complete remissions of both diseases by rituximab-bendamustine regimen combined to hypomethylating therapy. J Chemother. 2013; 25(4):247-9. doi: 10.1179/1973947813Y.0000000076.10. Gaul L, Weber S, Bauman P, Emmerich B, Schmidmaier R. Bendamustine induces G2 cell cycle arrest and apoptosis in myeloma cells: the role of ATM-Chk2-Cdc25A and ATM-p53-21 pathways. J. Cancer Res. Clin. Oncol. 2008; 134:245–253.11. Roue G, Lopez M, Milpied P et al. Bendamustine Is Effective in p53-Deficient B-Cell Neoplasms and Requires Oxidative Stress and Caspase-Independent Signaling Clin Cancer Res. 2008; 14:6907-6915.12. Schwanen C, Hecker T, Huabinger G, Woalfle M, Rittgen W, Bergmann L, Karakas T. In vitro evaluation of bendamustine induced apoptosis in B-chronic lymphocytic leukemia. Leukemia 2002; 16:2096–2105.13. Kaczmarek A, Vandenabeele P, Krysko DV. Necroptosis: The release of damage-associated molecular patterns and its physcological relevance. Immunity 2013; Feb 21.14. Fulda S. The mechanism of necroptosis in normal and cancer cells. Cancer Biology & Therapy 2013; 14:11, 999–1004.15. Leoni LM, Bailey B, Reifert J et al. Bendamustine (Treanda) displays a distinct pattern of cytotoxicity and unique mechanistic features compared with other alkylating agents. Clin. Cancer Res. 2008; 14:309–17.16. Cai B, Lyu H, Huang J, Wang S, Lee CK, Gao C, Liu B. Combination of bendamustine and entinostat synergistically inhibits proliferation of multiple myeloma cells via induction of apoptosis and DNA damage response. Cancer Lett. 2013; 28;335(2):343-50. doi: 10.1016/j.canlet.2013.02.046.
Abstract
References
- 1. Ozegowski W, Krebs D. w-[bis-(chlorethyl)-amino-benzimidazolyl-(2)]-propionic or butyric acids as potential cytostatic agents J Prakt Chem 1963; 20:178–86.2. Hartmann M, Zimmer CH. Investigation of cross-link formation in DNA by the alkylating cytostatica IMET 3106, 3393 and 3943 Biochim Biophys Acta 1972; 287:386–9.3. Strumberg D, Harstrick A, Doll K et al. Bendamustine hydrochloride activity against doxorubicine-resistant human breast cancer cell lines Anticancer Drugs 1996; 7:415–21.4. Staib P, Schinkothe T, Dimski T et al. In vitro modulation of ara-CTP accumulation in fresh AML cells by bendamustine in comparison with fludarabine, 2-CDA and gemcitabine Blood 1999; 94:63.5. Hiraoka N, Kikuchi J, Yamauchi T, Koyama D, Wada T, Uesawa M, Akutsu M, Mori S, Nakamura Y, Ueda T, Kano Y. Furukawa Purine analog-like properties of bendamustine underlie rapid activation of DNA damage response and synergistic effects with pyrimidine analogues in lymphoid malignancies Y.PLoS One 2014; 13;9(3):e90675. doi: 10.1371/journal.pone.0090675.6. Kost SE, Bouchard ED, LaBossière É, Ye X, Queau ML, Liang WS, Banerji V, Gibson SB, Katyal S,Johnston JB. Cross-resistance and synergy with bendamustine in chronic lymphocytic leukemia Leuk Res. 2016 ;50:63-71. doi: 10.1016/j.leukres.2016.09.016.7. Scutaru AM, Wenzel M, Scheffler H, Wolber G, Gust R. Optimization of the N-lost drugs melphalan and bendamustine: synthesis and cytotoxicity of a new set of dendrimer-drug conjugates as tumor therapeutic agents. Bioconjug Chem. 2010; 21(10):1728-43. doi: 10.1021/bc900453f.8. Inoue M, Honma Y, Urano T, Suzumiya J.Japanese apricot extract (MK615) potentiates bendamustine-induced apoptosis via impairment of the DNA damage response in lymphoma cells. Oncol Lett. 2017 Jul;14(1):792-800. doi: 10.3892/ol.2017.6219.9. Palombi M, Niscola P, Tendas A, Trawinska MM, Scaramucci L, Giovannini M, Fratoni S, Perrotti A, de Fabritiis P. Simultaneous occurrence of large B-cell non-Hodgkin lymphoma and acute myeloid leukaemia in an elderly patient: complete remissions of both diseases by rituximab-bendamustine regimen combined to hypomethylating therapy. J Chemother. 2013; 25(4):247-9. doi: 10.1179/1973947813Y.0000000076.10. Gaul L, Weber S, Bauman P, Emmerich B, Schmidmaier R. Bendamustine induces G2 cell cycle arrest and apoptosis in myeloma cells: the role of ATM-Chk2-Cdc25A and ATM-p53-21 pathways. J. Cancer Res. Clin. Oncol. 2008; 134:245–253.11. Roue G, Lopez M, Milpied P et al. Bendamustine Is Effective in p53-Deficient B-Cell Neoplasms and Requires Oxidative Stress and Caspase-Independent Signaling Clin Cancer Res. 2008; 14:6907-6915.12. Schwanen C, Hecker T, Huabinger G, Woalfle M, Rittgen W, Bergmann L, Karakas T. In vitro evaluation of bendamustine induced apoptosis in B-chronic lymphocytic leukemia. Leukemia 2002; 16:2096–2105.13. Kaczmarek A, Vandenabeele P, Krysko DV. Necroptosis: The release of damage-associated molecular patterns and its physcological relevance. Immunity 2013; Feb 21.14. Fulda S. The mechanism of necroptosis in normal and cancer cells. Cancer Biology & Therapy 2013; 14:11, 999–1004.15. Leoni LM, Bailey B, Reifert J et al. Bendamustine (Treanda) displays a distinct pattern of cytotoxicity and unique mechanistic features compared with other alkylating agents. Clin. Cancer Res. 2008; 14:309–17.16. Cai B, Lyu H, Huang J, Wang S, Lee CK, Gao C, Liu B. Combination of bendamustine and entinostat synergistically inhibits proliferation of multiple myeloma cells via induction of apoptosis and DNA damage response. Cancer Lett. 2013; 28;335(2):343-50. doi: 10.1016/j.canlet.2013.02.046.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Clinical Sciences |
| Journal Section | Research Article |
| Authors | |
| Publication Date | September 28, 2018 |
| Submission Date | April 18, 2018 |
| Acceptance Date | August 8, 2018 |
| Published in Issue | Year 2018 Volume: 11 Issue: 3 |
