Effect of miR-105-5p on Cell Proliferation, Colony Formation, and hTERT/P53/P21 Gene Expression in Multiple Cancer Cell Lines

Öz

Aim: MicroRNAs are small endogenous RNA molecules that do not code for proteins but have the capacity to influence cancer development by promoting or inhibiting it. They play normal biological roles in regulating cell metabolism, development, apoptosis, and metastasis. While numerous studies on the functional mechanisms of miRNAs exist, there is limited research in the literature on miR-105-5p, a member of the miR-105 family. This study investigated the effects of miR-105-5p on cell proliferation, colony formation, and the expression of hTERT, P21, and P53 genes in five cancer cell lines: A549 (lung), Huh7 (liver), HCT116 (colon), MCF-7 (breast), and HeLa (cervix).

Method: The expression of miR-105-5p was both upregulated and downregulated through transfection in the selected cancer cell lines. After transfection, assays were performed to assess cell proliferation, colony-forming ability, and the expression of the hTERT, P21, and P53 genes.

Results: Overexpression of miR-105-5p inhibited proliferation in all cancer cell lines. miR-105-5p suppressed colony formation in A549, MCF-7, HeLa, and Huh7 cells. Differences in the expression levels of hTERT, P53, and P21 genes were observed in different cell lines due to the effect of the miR-105-5p mimic.

Conclusion: Direct or indirect differences in hTERT gene expression levels suggest that miR-105-5p may be useful in targeted therapy.

Anahtar Kelimeler

• miR-105-5p
• hTERT
• P53
• P21
• transfection

Destekleyen Kurum

Çukurova Üniversitesi

Proje Numarası

FYL-2021-14344, TYL-2021-13415

Etik Beyan

In this study, we affirm that all regulations outlined in the "Higher Education Institutions Scientific Research and Publication Ethics Directive" have been strictly followed. Furthermore, no actions contrary to the principles described under "Actions Against Scientific Research and Publication Ethics" have been undertaken.

Teşekkür

We would like to thank the Scientific Research Unit of Çukurova University for their support in the conduct of this study with projects numbered FYL-2021-14344 and BAP: TYL-2021-13415. We would also like to thank the Central Laboratory and Biotechnology Center of Cukurova University for providing space and equipment for conducting of the experimental stages of the project.

miR-105-5p'nin Çeşitli Kanser Hücre Hatlarında Hücre Proliferasyonu, Koloni Oluşumu ve hTERT/P53/P21 Gen Ekspresyonuna Etkisi

Öz

Amaç: MikroRNA'lar, protein kodlamayan ancak kanser gelişimini teşvik ederek veya engelleyerek etkileme kapasitesine sahip küçük endojen RNA molekülleridir. Hücre metabolizması, gelişim, apoptoz ve metastazın düzenlenmesinde normal biyolojik roller oynarlar. MikroRNA'ların fonksiyonel mekanizmalarına ilişkin çok sayıda çalışma mevcut olmasına rağmen, miR-105 ailesinin bir üyesi olan miR-105-5p hakkında literatürde sınırlı araştırma bulunmaktadır. Bu çalışma, beş kanser hücresi hattında (A549 (akciğer), Huh7 (karaciğer), HCT116 (kolon), MCF-7 (meme) ve HeLa (serviks)) miR-105-5p'nin hücre çoğalması, koloni oluşumu ve hTERT, P21 ve P53 genlerinin ekspresyonu üzerindeki etkilerini araştırmıştır.

Yöntem: Seçilen kanser hücre hatlarında miR-105-5p ekspresyonu, transfeksiyon yoluyla hem yukarı hem de aşağı doğru düzenlenmiştir. Transfeksiyondan sonra, hücre çoğalması, koloni oluşturma yeteneği ve hTERT, P21 ve P53 genlerinin ekspresyonunu değerlendirmek için analizler yapıldı.

Bulgular: miR-105-5p'nin aşırı ekspresyonu tüm kanser hücre hatlarında çoğalmayı inhibe etti. miR-105-5p, A549, MCF-7, HeLa ve Huh7 hücrelerinde koloni oluşumunu baskıladı. miR-105-5p taklidinin etkisi nedeniyle farklı hücre hatlarında hTERT, P53 ve P21 genlerinin ekspresyon seviyelerinde farklılıklar gözlendi.

Sonuç: hTERT gen ekspresyon seviyelerindeki doğrudan veya dolaylı farklılıklar, miR-105-5p'nin hedefli tedavide yararlı olabileceğini düşündürmektedir.

Anahtar Kelimeler

• miR-105-5p
• hTERT
• P53
• P21
• transfeksiyon

Destekleyen Kurum

Çukurova Üniversitesi Bilimsel Araştırma Birimi

Proje Numarası

FYL-2021-14344, TYL-2021-13415

Etik Beyan

Bu çalışmada, "Yükseköğretim Kurumları Bilimsel Araştırma ve Yayın Etiği Yönergesi"nde belirtilen tüm düzenlemelere harfiyen uyulduğunu ve "Bilimsel Araştırma ve Yayın Etiğine Aykırı Eylemler" başlığı altında açıklanan ilkelere aykırı herhangi bir eylemde bulunulmadığını beyan ederiz.

Teşekkür

Bu çalışmanın yürütülmesinde FYL-2021-14344 ve BAP: TYL-2021-13415 numaralı projelerle bize destek olan Çukurova Üniversitesi Bilimsel Araştırma Birimi'ne teşekkür ederiz. Ayrıca, projenin deneysel aşamalarının yürütülmesi için mekan ve ekipman sağlayan Çukurova Üniversitesi Merkez Laboratuvarı ve Biyoteknoloji Merkezi'ne de teşekkür ederiz.

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