İnsan Akciğer Karsinomu Hücre Hattında LncRNA PVT1 ve MiR-128'in İfade Düzeylerinin Belirlenmesi

Abstract

Amaç: Araştırmamızın amacı, normal akciğer epitel hücre hatlarında (BEAS-2B) ve insan akciğer karsinomunda (A549), mikroRNA-128 (miR-128) ve uzun kodlamayan RNA (lncRNA) PVT1 ekspresyonunu inceleyerek, bu moleküllerin akciğer kanserinin gelişimi ve ilerlemesinde oynayabileceği potansiyel rolleri belirlemektir. Gereç ve Yöntemler: Bu araştırmada kullanılan hücre hatları A549 ve BEAS-2B'dir. Gerçek zamanlı kantitatif PCR (RT-qPCR) yöntemi kullanılarak PVT1 ve miR-128'in ifade seviyeleri değerlendirildi. 2-∆∆Ct yöntemi, U6 snRNA ve GAPDH ifadelerine göre normalize ederek kat değişimlerini incelemek için kullanıldı. Bulgular: Akciğer kanseri hücrelerinde PVT1 ekspresyonunun yükseldiği belirlendi (p<0,01). Bununla birlikte, miR-128 ekspresyonu istatistiksel olarak anlamlı bir şekilde değişmedi (p>0,05). Mir-128, in-silico analizde lncRNA PVT1'in potansiyel bir bağlanma hedefi olmasına rağmen, ekspresyon profili sonuçlarına göre korelasyon görülmedi. Sonuç: Normal akciğer epitel hücrelerine kıyasla akciğer kanseri hücrelerinde lncRNA PVT1 ve miR-128 arasındaki ilişkinin incelendiği çalışmamız, PVT1 ekspresyonunun akciğer kanseri hücrelerinde önemli ölçüde yüksek olduğunu, buna karşın miR-128 ekspresyonunun değişmeden kaldığını ortaya koymaktadır. LncRNA PVT1 ve miR-128 arasındaki etkileşimin dolaylı olup olmadığını keşfetmek için daha fazla çalışmaya ihtiyaç vardır.

Keywords

• Akciğer kanseri
• A549 hücre hattı
• PVT1
• miR-128
• RT-qPCR

Determining the Expression Levels of LncRNA PVT1 and MiR-128 in the Human Lung Carcinoma Cell Line

Abstract

Objective: In the presented research, we aimed to examine the expression of microRNA-128 (miR-128) and long non-coding RNA (lncRNA) the plasmacytoma variant translocation 1 (PVT1) in non-cancerous cells BEAS-2B and human lung carcinoma A549 to identify any potential roles these molecules might play in the cancer development in lung tissue. Material and Methods: The cell lines used in this research were A549 and BEAS-2B. Total RNA was isolated from these cell lines, and then complementary DNA (cDNA) was synthesised. Using the quantitative real-time PCR (RT-qPCR) method, the expression levels of lncRNA PVT1 and miR-128 were evaluated. The 2-∆∆Ct method was employed to analyse fold changes by normalising to U6 snRNA and GAPDH expressions. Results: Elevated PVT1 expression was determined in the lung cancer cells (p<0.01). Nevertheless, miR-128 expression did not change statistically significantly (p>0.05). Even though miR-128 is a potential binding target of lncRNA PVT1 according to in silico analysis, the expression profile does not correlate. Conclusion: Our evaluation of the relationship between lncRNA PVT1 and miR-128 in cancerous cells A549 compared with non-cancerous cells BEAS-2B reveals that PVT1 expression is significantly elevated in lung cancer cells, whereas miR-128 expression remains unchanged. Further investigation is needed to explore whether the interaction between lncRNA PVT1 and miR-128 is indirect.

Keywords

• Lung cancer
• A549 cell line
• PVT1
• miR-128
• RT-qPCR

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