Impact of KLF4, SHH, and hif1a knockdown on miRNA expression in malign melanoma cancer stem cells

Berrin Ozdil; Çığır Biray Avcı; Hüseyin Aktuğ

doi:10.19161/etd.1528915

EN TR

Impact of KLF4, SHH, and hif1a knockdown on miRNA expression in malign melanoma cancer stem cells

Abstract

Aim: microRNAs (miRNAs) play a pivotal role in gene regulation, influencing various cellular processes such as differentiation, proliferation, and apoptosis. This study investigated the expression of three specific miRNAs (Hsa-miR-21-5p, Hsa-miR-9-5p, and Hsa-miR-200a-5p) in malign melanoma stem cells (CSCs) and non-stem cells (NSCs). Method: CSCs and NCSCs were sorted from CHL-1 cells based on CD133 marker, a malignant melanoma cell line. CD133+ cells were treated with Hif1a, KLF4, and SHH siRNA and the expression levels of three different miRNAs were compared between groups. Results: Our results revealed that Hsa-miR-200a-5p expression was similar in both cell groups. Conversely, Hsa-miR-21-5p and Hsa-miR-9-5p were significantly upregulated in NCSCs. Further analysis showed that the knockdown of KLF4 did not significantly affect the expression levels of these miRNAs. However, silencing SHH resulted in a substantial downregulation of Hsa-miR-21-5p and a significant upregulation of Hsa-miR-9-5p. Additionally, Hif1a knockdown led to the downregulation of both Hsa-miR-21-5p and hsa-miR-9-5p. Conclusion: These findings highlight the complex regulatory mechanisms of miRNA expression in different cellular contexts and suggest potential roles for these miRNAs in response to specific gene silencing.

Keywords

KLF4, SHH ve Hif1a Susturulmasının Malign Melanom Kanser Kök Hücrelerinde miRNA Ekspresyonuna Etkisi

Öz

Amaç: mikroRNA'lar (miRNA'lar) gen regülasyonunda önemli bir rol oynar ve farklılaşma, proliferasyon ve apoptoz gibi çeşitli hücresel süreçleri etkiler. Bu çalışmada, malign melanom kök hücrelerinde (CSC'ler) ve kök hücre olmayan hücrelerde (NSC'ler) üç spesifik miRNA'nın (Hsa-miR-21-5p, Hsa-miR-9-5p ve Hsa-miR-200a-5p) ekspresyonu araştırılmıştır. Yöntem: Malign melanoma hücre hattı olan CHL-1 hücrelerinden CSC ve NCSC hücreleri CD133 belirteci baz alınarak elde edilmiştir. CD133+ hücreler HIF1a, KLF4, ve SHH siRNA ile muamele edilerek üç farklı miRNA ekspresyon seviyesi gruplar arası karşılaştırılmıştır. Bulgular: Bulgularımız Hsa-miR-200a-5p ekspresyonunun her iki hücre grubunda da benzer olduğunu ortaya koymuştur. Buna karşılık, Hsa-miR-21-5p ve Hsa-miR-9-5p, NCSC hücrelerinde önemli ölçüde yüksek ifade edilmiştir. Daha ileri analizler KLF4'ün susturulmasının bu miRNA'ların ifade düzeylerini önemli ölçüde etkilemediğini göstermiştir. Bununla birlikte, SHH'nin susturulması Hsa-miR-21-5p'nin önemli ölçüde düşük regülasyonu ve Hsa-miR-9-5p'nin önemli ölçüde yüksek ifadelenmesi ile sonuçlanmıştır. Ek olarak, Hif1a'nın susturulması hem Hsa-miR-21-5p hem de Hsa-miR-9-5p'nin aşağı regülasyonuna yol açmıştır. Sonuç: Elde edilen bulgular, farklı hücresel bağlamlarda miRNA ifadesinin karmaşık düzenleyici mekanizmalarını vurgulamakta ve spesifik gen susturmaya yanıt olarak bu miRNA'lar için potansiyel roller önermektedir.

Anahtar Kelimeler

Supporting Institution

Scientific Research Projects Coordination Unit at Ege University

Project Number

20785

Thanks

The authors thank to the Scientific Research Projects Coordination Unit at Ege University for their financial support.

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Details

Primary Language

English

Subjects

Cancer Cell Biology , Molecular Targets

Journal Section

Research Article

Authors

Berrin Ozdil ^*
0000-0001-6081-2308
Türkiye

Çığır Biray Avcı
0000-0001-8251-4520
Türkiye

Hüseyin Aktuğ
0000-0003-4150-8495
Türkiye

Publication Date

December 9, 2024

Submission Date

August 6, 2024

Acceptance Date

September 11, 2024

Published in Issue

Year 1970 Volume: 63 Number: 4

DOI

https://doi.org/10.19161/etd.1528915

IZ

https://izlik.org/JA75JD39HU

Cite

RIS / Bibtex

Vancouver

1.Berrin Ozdil, Çığır Biray Avcı, Hüseyin Aktuğ. Impact of KLF4, SHH, and hif1a knockdown on miRNA expression in malign melanoma cancer stem cells. EJM. 2024 Dec. 1;63(4):595-602. doi:10.19161/etd.1528915

Cited By

Integrated spectroscopic and morphological analyses reveal cellular shifts in gene-silenced melanoma CSCs

Scientific Reports

https://doi.org/10.1038/s41598-025-17155-2