Prostat Kanseri Hücre Hatlarında lncRNA H19 ve miR-29b-3p Ekspresyon Profilleri

Abstract

Amaç: Bu çalışmada, insan prostat kanseri hücre hatlarında (PC3, DU145, LNCaP) ve normal prostat epitel hücrelerinde (PNT1A) uzun olmayan kodlayıcı RNA H19 ve mikroRNA-29b-3p (miR-29b-3p) düzeylerinin incelenmesi ve bu moleküllerin prostat kanseri biyolojisindeki potansiyel rolleri ve etkileşimlerinin değerlendirilmesi amaçlanmıştır. Gereç ve Yöntem: PC3, DU145, LNCaP ve PNT1A hücre hatları belirlenmiş protokole göre kültüre edilmiştir. Hücreler konfluansa ulaştığında toplam RNA ekstrakte edilmiştir. H19 ve miR-29b-3p ekspresyon düzeyleri RT-qPCR yöntemi ile ölçülmüş ve sırasıyla GAPDH ve U6 snRNA’ya göre normalize edilmiştir. Göreli gen ekspresyon düzeylerini belirlemek için 2-ΔΔCt yöntemi uygulanmıştır. Bulgular: lncRNA H19 ve miR-29b-3p düzeylerindeki değişiklikler, tüm kanser hücre hatlarında PNT1A hücrelerine kıyasla istatistiksel olarak anlamlı bulunmuştur. H19 düzeyleri PC3'te belirgin şekilde yüksek, DU145’te orta derecede artmış, LNCaP'ta ise anlamlı şekilde azalmıştır. Tersine, miR-29b-3p düzeyleri DU145 ve LNCaP'ta artarken, PC3’te belirgin şekilde azalmıştır. İstatistiksel analizler özellikle H19 için PC3 ile diğer hücre hatları arasında ve miR-29b-3p için PC3 ile DU145 arasında anlamlı farklılıklar olduğunu göstermiştir (p<0,05). Sonuç: Bulgularımız, PC3 hücrelerinde H19 ile miR-29b-3p arasında negatif bir korelasyon olduğunu göstermektedir; bu durum, H19’un miR-29b-3p için moleküler bir sünger görevi görebileceğini düşündürmektedir. LNCaP ile diğerleri arasında gözlemlediğimiz ekspresyon düzeyi farklılıkları, H19–miR-29b-3p ekseninin androjen reseptörü sinyallemesi ve tümör agresifliğinden etkilenebileceğini göstermektedir. Altta yatan mekanizmayı ve olası klinik önemini daha iyi anlayabilmek için ilave fonksiyonel çalışmalara ihtiyaç vardır.

Keywords

• Prostat kanseri
• lncRNA H19
• miR-29b-3p
• Gen ekspresyonu
• ceRNA ağı
• Tümör progresyonu

Expression Profiles of lncRNA H19 and miR-29b-3p in Prostate Cancer Cell Lines

Abstract

Objective: We aimed to evaluate the expression levels of long non-coding RNA H19 and microRNA-29b-3p (miR-29b-3p) in human prostate cancer cell lines (PC3, DU145, LNCaP) and in normal prostate epithelial cells (PNT1A) to explore their potential roles and interactions in prostate cancer biology. Materials and Methods: All cell lines PC3, DU145, LNCaP, and PNT1A were cultured in the same type of medium, RPMI-1640. Total RNA was extracted from the cells once they reached confluency. H19 and miR-29b-3p levels were measured by real-time quantitative PCR and normalised to GAPDH and U6 snRNA, respectively. Their relative level was determined by the 2-ΔΔCt method. Results: The alterations in lncRNA H19 and miR-29b-3p levels were statistically significant across all cancer cell lines compared to PNT1A cells. H19 levels were notably higher in PC3, moderately increased in DU145, and significantly decreased in LNCaP. Conversely, miR-29b-3p levels increased in DU145 and LNCaP but decreased markedly in PC3. Statistical analysis particularly showed significant differences between PC3 and other cell lines for lncRNA H19 and between PC3 and DU145 for miR-29b-3p (p<0.05). Conclusion: Our findings show an inverse relationship between H19 and miR-29b-3p in PC3 cells, indicating that H19 may function as a molecular sponge for miR-29b-3p. The differences we observed in the expression levels between LNCaP and the others show that the H19–miR-29b-3p axis may be affected by androgen receptor signalling and tumour aggressiveness. To better understand the underlying mechanism and its potential clinical relevance, additional functional studies are necessary.

Keywords

• Prostate cancer
• lncRNA H19
• miR-29b-3p
• Gene expression
• ceRNA network
• Tumour progression

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