The effect of miR-640 on metastatic genes in breast cancer

İsmail Mert Alkaç; Çığır Biray Avcı

doi:10.19161/etd.1771783

EN TR

The effect of miR-640 on metastatic genes in breast cancer

Abstract

Aim: Despite the implementation of numerous treatment protocols for breast cancer, which remains a leading cause of cancer-related mortality among women, metastasis continues to pose a major challenge during therapy. This results in prolonged treatment durations, poor prognosis, and the need to manage newly emerging cancer types due to the development of secondary tumors. This study aimed to investigate the anti-metastatic effects of miR-640 previously shown to influence tumorigenesis, angiogenesis, inflammatory responses, and metastasis in various cancers, including breast cancer—by examining its potential to downregulate metastasis-associated genes. Materials and Methods: To assess the anti-metastatic potential of miR-640, functional assays—including wound healing, Annexin V, analyses—were conducted following lipofectamine-mediated transfection in MCF7 and MDA-MB-231 breast cancer cell lines. Additionally, expression levels of metastasis-associated genes (WNT7A, TMSB4X, NFIB, ZEB2, IL11, AURKA, CAV1, SLC7A11, PDK1, and PTK6) identified as potential targets of miR-640, were analyzed. Results: Successful transfection was achieved in both cell lines, with a significant increase in miR-640 expression (~25-fold in MCF7, ~5-fold in MDA-MB-231). Statistically significant effects of miR-640 were observed in wound healing and Annexin V assays following transfection. Additionally, gene expression analyses revealed significant reductions in the expression of metastasis-related genes WNT7A, TMSB4X, NFIB, IL11, AURKA, CAV1, SLC7A11, PDK1, PTK6 in the MCF7 cell line, and in WNT7A, NFIB and ZEB2 genes in the MDA-MB-231 cell line. Conclusion: This study is the first to demonstrate that miR-640 exerts an anti-metastatic effect in breast cancer by suppressing metastatic formation through the downregulation of metastasis-related genes. The findings show that cell migration is significantly inhibited in the transfected cell lines and suggest that miR-640 may serve as a potential therapeutic molecule

Keywords

miR-640'ın meme kanserinde metastatik genler üzerindeki etkisi

Öz

Amaç: Kadınlarda kansere bağlı ölümlerin başlıca nedeni olmaya devam eden meme kanseri için çok sayıda tedavi protokolü uygulanmasına rağmen, metastaz tedavi sırasında önemli bir sorun olmaya devam etmektedir. Bu durum, tedavi süresinin uzamasına, kötü prognoza ve ikincil tümörlerin gelişmesi nedeniyle yeni ortaya çıkan kanser türlerinin yönetilmesi ihtiyacına yol açmaktadır. Bu çalışma, daha önce meme kanseri dahil çeşitli kanserlerde tümör oluşumu, anjiyogenez, inflamatuar yanıtlar ve metastazı etkilediği gösterilen miR-640'ın metastazla ilişkili genleri aşağı regüle etme potansiyelini inceleyerek, anti-metastatik etkilerini araştırmayı amaçlamıştır. Gereç ve Yöntem: miR-640'ın anti-metastatik potansiyelini değerlendirmek için, MCF7 ve MDA-MB-231 meme kanseri hücre hatlarında lipofektamin aracılı transfeksiyonun ardından yara iyileşmesi, Annexin V ve hücre döngüsü analizleri dahil olmak üzere fonksiyonel testler gerçekleştirilmiştir. Ek olarak, miR-640'ın potansiyel hedefleri olarak tanımlanan metastazla ilişkili genlerin (WNT7A, TMSB4X, NFIB, ZEB2, IL11, AURKA, CAV1, SLC7A11, PDK1 ve PTK6) ekspresyon düzeyleri analiz edildi. Bulgular: Her iki hücre hattında da başarılı transfeksiyon gerçekleştirildi ve miR-640 ekspresyonunda önemli bir artış gözlendi (MCF7'de ~25 kat, MDA-MB-231'de ~5 kat). Transfeksiyonun ardından yara iyileşmesi ve Annexin V testlerinde miR-640'ın istatistiksel olarak anlamlı etkileri gözlendi. Ek olarak, gen ekspresyon analizleri, MCF7 hücre hattında metastazla ilişkili genler WNT7A, TMSB4X, NFIB, IL11, AURKA, CAV1, SLC7A11, PDK1, PTK6'nın ve MDA-MB-231 hücre hattında WNT7A, NFIB ve ZEB2 genlerinin ekspresyonunda önemli azalmalar olduğunu ortaya koydu. Sonuç: Bu çalışma, miR-640'ın metastazla ilişkili genlerin aşağı regülasyonu yoluyla metastaz oluşumunu baskılayarak meme kanserinde anti-metastatik etki gösterdiğini ortaya koyan ilk çalışmadır. Bulgular, transfekte edilmiş hücre hatlarında hücre göçünün önemli ölçüde inhibe edildiğini göstermektedir ve miR-640'ın potansiyel bir terapötik molekül olarak hizmet edebileceğini düşündürmektedir.

Anahtar Kelimeler

Supporting Institution

This study was supported by the TUSEB (Presidency of Turkish Health Institutes) A GROUP EMERGENCY Research and Development PROJECT SUPPORT PROGRAM No. 35685.

Project Number

35685

Thanks

This study was supported by the TUSEB (Presidency of Turkish Health Institutes) A GROUP EMERGENCY Research and Development PROJECT SUPPORT PROGRAM No. 35685.

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Details

Primary Language

English

Subjects

Cancer Therapy (Excl. Chemotherapy and Radiation Therapy) , Chemotherapy , Molecular Targets , Oncology and Carcinogenesis (Other)

Journal Section

Research Article

Authors

İsmail Mert Alkaç ^*
0000-0003-0847-7738
Türkiye

Çığır Biray Avcı
0000-0001-8251-4520
Türkiye

Publication Date

March 9, 2026

Submission Date

August 25, 2025

Acceptance Date

September 10, 2025

Published in Issue

Year 1970 Volume: 65 Number: 1

DOI

https://doi.org/10.19161/etd.1771783

IZ

https://izlik.org/JA85MA83PB

Cite

RIS / Bibtex

Vancouver

1.İsmail Mert Alkaç, Çığır Biray Avcı. The effect of miR-640 on metastatic genes in breast cancer. EJM. 2026 Mar. 1;65(1):1-10. doi:10.19161/etd.1771783