E-ISSN: 1308-5263
Ortho-Topolin Riboside Induced Differentiation through Inhibition of STAT3 Signaling in Acute Myeloid Leukemia HL-60 Cells [Turk J Hematol]
Turk J Hematol. 2019; 36(3): 162-168 | DOI: 10.4274/tjh.galenos.2019.2019.0020  

Ortho-Topolin Riboside Induced Differentiation through Inhibition of STAT3 Signaling in Acute Myeloid Leukemia HL-60 Cells

Li Wang1, Jiao Cheng1, Fanlin Lin1, Shengxian Liu1, Hui Pan1, Mingda Li1, Shanshan Li1, Na Li2, Weiping Li2
1School of Life and Medicine, Dalian University of Technology, PanJin, China
2The Second Hospital of Dalian Medical University, Dalian, China

Objective: We previously demonstrated that ortho-topolin riboside (oTR) as a naturally occurring cytokinin secreted from Populus × robusta has great potential anticancer effects via the mitochondrial apoptotic pathway and endoplasmic reticulum stress pathway. In the present study, we reveal that oTR induced the differentiation of acute myeloid leukemia (AML) HL-60 cells, which represent the M2 subtype of AML.
Materials and Methods: After the incubation of HL-60 cells with oTR, its effect was analyzed with cell viability assay, Wright-Giemsa staining, CD11b protein expression analysis, western blot analysis, and polymerase chain reaction.
Results: We found that oTR arrested the cell cycle at the S phase, upregulated the expression of myeloid surface marker CD11b, reduced the nuclear cytoplasmic ratio, and altered the horseshoe shape of nuclei, as evidenced by Wright-Giemsa staining. Furthermore, we found that the protein level of phosphorylated STAT3 was decreased when cells were treated with oTR, while phosphorylated STAT1 was activated. Moreover, the protein level of phosphorylated STAT3 and its upstream kinase, Janus kinase 2, were also inhibited when cells were treated with oTR after increased time. Additionally, the levels of phosphorylated SHP-1 were increased while phosphorylated SHP-2 was decreased.
Conclusion: Collectively, our data indicate a differentiation-induced mechanism underlying the inhibition of STAT3 signaling upon treatment with oTR. Therefore, oTR may constitute a novel differentiation-induced therapeutic for use in clinical treatment of AML.

Keywords: Ortho-topolin riboside, Differentiation, STAT3 signal, HL-60 cells


Akut Myeloid Lösemi HL-60 Hücre Serisinde Orto-Topolin Ribozid ile Diferansiyasyonun STAT3 Sinyal İnhibisyonu Yoluyla İndüklenmesi

Li Wang1, Jiao Cheng1, Fanlin Lin1, Shengxian Liu1, Hui Pan1, Mingda Li1, Shanshan Li1, Na Li2, Weiping Li2
1School of Life and Medicine, Dalian University of Technology, PanJin, China
2The Second Hospital of Dalian Medical University, Dalian, China

Amaç: Daha önceki çalışmalarımızda Populus × robusta’dan doğal olarak oluşan sitokin orto-topolin ribozidin (oTR) mitokondrial apopitotik yolaklar ve endoplazmik retikulum stres yolakları vasıtasıyla önemli bir antikanser potansiyelinin olduğunu göstermiştik. Bu çalışmada, oTR’nin AML M2 subtipi özelliğindeki HL-60 hücre serisinde diferansiyasyonu indüklediğini gösterdik.
Gereç ve Yöntem: HL-60 hücrelerinin oTR ile inkübasyonunu takiben hücre üzerideki etkileri hücre canlılık testleri, Wright-Giemsa boyaması, CD11b protein ekspresyon analizi, western blot analizi ve polimeraz zincir reaksiyonu ile araştırıldı.
Bulgular: oTR’nin hücre siklusunun S fazında duraklattığını, myeloid hücre yüzey belirteçlerinden CD11b ekspresyonunun arttığını, çekirdek sitoplazma oranının azaldığını ve çekirdeğin atnalı şeklinin değiştiğini Wright-Giemsa boyası ile destekleyerek gördük. oTR ile muamele edilmiş hücrelerde fosforile STAT3 protein düzeyinin azaldığını, fosforile STAT1’in ise aktive olduğunu bulduk. Ayrıca fosforile STAT3 ve yukarı yöndeki kinaz olan Janus kinaz 2’nin, hücrelerin oTR ile inkübasyonunda artmış zamanla inhibe olduğu görüldü. Ek olarak fosforile SHP-1 düzeyleri artarken fosforile SHP-2 düzeyi azaldı.
Sonuç: Birlikte değerlendirildiğinde, sonuçlarımız oTR ile STAT3 inhibisyonu üzerinden bir diferansiyasyon indükleme mekanizmasını işaret etmektedir. Bu nedenle, oTR AML tedavisinde yeni bir diferansiyasyon-indükleyici terapötik olarak yer alabilir.

Anahtar Kelimeler: Orto-topolin ribozid, Diferansiyasyon, STAT3 sinyali, HL-60 hücreler


Li Wang, Jiao Cheng, Fanlin Lin, Shengxian Liu, Hui Pan, Mingda Li, Shanshan Li, Na Li, Weiping Li. Ortho-Topolin Riboside Induced Differentiation through Inhibition of STAT3 Signaling in Acute Myeloid Leukemia HL-60 Cells. Turk J Hematol. 2019; 36(3): 162-168

Corresponding Author: Weiping Li, Türkiye


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