Dual Role of Colchicine in Glioblastoma: miR-22-3p-Dependent Regulation of Wound Healing and Proliferation

Abstract

ABSTRACT Objective: The downregulation of miR-22-3p in glioblastoma (GBM) tissues and cells has been shown to plays a critical role in gliomagenesis and patient prognosis. However, the effects of colchicine on GBM cellular functions via miR-22-3p mediated mechanisms remain unexplored. This study aims to investigate the effects of colchicine on GBM cell behavior through miR-22-3p-mediated mechanisms. Methods: miR-22-3p expression levels were quantified in U87 GBM cells treated with 1, 10, and 100 ng/ml colchicine using qRT-PCR. Cell viability and drug resistance were evaluated using the CCK-8 assay across four groups: temozolomide (TMZ) treated or untreated (i) U87 cells (control), (ii) U87 cells treated with 10 ng/ml colchicine, (iii) U87 cells transfected with a miR-22-3p inhibitor, and (iv) U87 cells transfected with a miR-22-3p inhibitor and 10 ng/ml colchicine. Colony formation was analyzed, migration was assessed using a transwell assay, and wound healing capacity was evaluated via scratch assay, all in U87 cells treated with 10 ng/ml colchicine. Results: Colchicine at 10 ng/ml significantly upregulated miR-22-3p expression. Inhibition of miR-22-3p reversed the colchicine-induced reduction in cell viability and colony formation. Neither colchicine nor miR-22-3p affected the resistance of U87 cells to TMZ. Colchicine reduced U87 cell migration independently of miR-22-3p, while its effect on wound healing was dependent on miR-22-3p expression. Conclusion: This study demonstrates that colchicine modulates miR-22-3p expression and influences key GBM cell functions. These findings provide a preliminary evidence supporting the potential of colchicine as a therapeutic agent in glioblastoma treatment.

Keywords

• Glioblastoma
• colchicine
• miR-22-3p
• transfection
• proliferation
• migration
• wound healing

Glioblastoma Hücrelerinde Kolşisinin Çift Etkisi: miR-22-3p Aracılı Yara İyileşmesi ve Hücre Proliferasyonunun Düzenlenmesi

Öz

ÖZ Amaç: Glioblastoma (GBM) doku ve hücrelerinde miR-22-3p'nin regülasyonu, gliomagenez ve hasta prognozunda kritik bir rol oynamaktadır. Ancak kolçisinin, GBM hücresel fonksiyonları üzerindeki miR-22-3p aracılı mekanizmalar yoluyla olan etkileri henüz araştırılmamıştır. Bu çalışmanın amacı, kolçisinin GBM hücre davranışları üzerindeki etkilerini miR-22-3p aracılı mekanizmalar üzerinden incelemektir. Yöntemler: U87 GBM hücrelerinde 1, 10 ve 100 ng/ml kolçisin uygulaması sonrasında miR-22-3p ekspresyon düzeyleri qRT-PCR ile ölçülmüştür. Hücre canlılığı ve ilaç direnci, CCK-8 testi kullanılarak dört grup üzerinden değerlendirilmiştir: temozolomid (TMZ) uygulanan ya da uygulanmayan (i) kontrol grubu U87 hücreleri, (ii) 10 ng/ml kolçisinle muamele edilen U87 hücreleri, (iii) miR-22-3p inhibitörü ile transfekte edilen U87 hücreleri ve (iv) hem miR-22-3p inhibitörü ile transfekte edilen hem de 10 ng/ml kolçisinle muamele edilen U87 hücreleri. Koloni oluşumu, koloni oluşum testi ile; migrasyon, transwell testi ile; yara iyileşmesi kapasitesi ise çizik (scratch) testi ile, 10 ng/ml kolçisin uygulanan U87 hücrelerinde analiz edilmiştir. Bulgular: 10 ng/ml kolçisin, U87 hücrelerinde miR-22-3p ekspresyonunu anlamlı düzeyde artırmıştır. miR-22-3p inhibisyonu, kolçisinin hücre canlılığı ve koloni oluşumu üzerindeki baskılayıcı etkisini tersine çevirmiştir. Kolçisin veya miR-22-3p, U87 hücrelerinin TMZ’ye olan ilaca direncini etkilememiştir. Kolçisin, hücre migrasyonunu miR-22-3p’den bağımsız olarak azaltırken, yara iyileşmesi üzerindeki etkisi miR-22-3p ekspresyonuna bağımlı olarak gözlenmiştir. Sonuç: Bu çalışma, kolçisinin miR-22-3p ekspresyonunu düzenlediğini ve glioblastoma hücrelerinin temel fonksiyonlarını etkilediğini göstermektedir. Bulgular, kolçisinin glioblastoma tedavisinde potansiyel bir terapötik ajan olarak değerlendirilmesine yönelik ön kanıtlar sunmaktadır.

Anahtar Kelimeler

• Glioblastoma
• kolşisin
• miR-22-3p
• transfeksiyon
• proliferasyon
• migrasyon
• yara iyileşmesi

Kaynakça

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