Determining the Expression Levels of LncRNA PVT1 and MiR-128 in the Human Lung Carcinoma Cell Line
Year 2025,
Volume: 8 Issue: 2, 69 - 75, 30.06.2025
Ezgi Nurdan Yenilmez Tunoğlu
,
Gamze Nur Öter
,
Remzi Okan Akar
,
Engin Ulukaya
,
Sevda Tanrıkulu Küçük
,
Fatma Hande Karpuzoğlu
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.
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İnsan Akciğer Karsinomu Hücre Hattında LncRNA PVT1 ve MiR-128'in İfade Düzeylerinin Belirlenmesi
Year 2025,
Volume: 8 Issue: 2, 69 - 75, 30.06.2025
Ezgi Nurdan Yenilmez Tunoğlu
,
Gamze Nur Öter
,
Remzi Okan Akar
,
Engin Ulukaya
,
Sevda Tanrıkulu Küçük
,
Fatma Hande Karpuzoğlu
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.
References
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- Seo D, Kim D, Chae Y, Kim W. The ceRNA network of lncRNA and miRNA in lung cancer. Genomics Inform 2020;18(4). google scholar
- Liu B, Li J, Cairns MJ. Identifying miRNAs, targets, and functions. Brief Bioinform 2014;15(1):1-19. google scholar
- Peng Y, Croce CM. The role of MicroRNAs in human cancer. Signal Transduct Target Ther 2016;1(1):1-9. google scholar
- Qian Y, Shi L, Luo Z. Long non-coding RNAs in cancer: implications for diagnosis, prognosis, and therapy. Front Med (Lausanne) 2020;7:612393. google scholar
- Wang K, Jin W, Song Y, Fei X. LncRNA RP11-436H11. 5, functioning as a competitive endogenous RNA, upregulates BCL-W expression by sponging miR-335-5p and promotes proliferation and invasion in renal cell carcinoma. Mol Cancer 2017;16:1-11. google scholar
- Yan X, Hu Z, Feng Y, Hu X, Yuan J, Zhao SD, et al. Comprehensive genomic characterization of long non-coding RNAs across human cancers. Cancer Cell 2015;28(4):529-40. google scholar
- Entezari M, Taheriazam A, Orouei S, Fallah S, Sanaei A, Hejazi ES, et al. LncRNA-miRNA axis in tumor progression and therapy response: An emphasis on molecular interactions and therapeutic interventions. Biomed Pharmacother 2022;154:113609. google scholar
- Chen J, Yu Y, Li H, Hu Q, Chen X, He Y, et al. Long non-coding RNA PVT1 promotes tumor progression by regulating the miR-143/HK2 axis in gallbladder cancer. Mol Cancer 2019;18:1-16. google scholar
- He YI, Jing Y, Wei F, Tang Y, Yang L, Luo J, et al. Long non-coding RNA PVT1 predicts poor prognosis and induces radioresistance by regulating DNA repair and cell apoptosis in nasopharyngeal carcinoma. Cell Death Dis 2018;9(2):235. google scholar
- Fu C, Li D, Zhang X, Liu N, Chi G, Jin X. LncRNA PVT1 facilitates tumorigenesis and progression of glioma via regulation of MiR-128-3p/GREM1 axis and BMP signaling pathway. Neurotherapeutics 2018;15:1139-57. google scholar
- Liu S, Chen W, Hu H, Zhang T, Wu T, Li X, et al. Long noncoding RNA PVT1 promotes breast cancer proliferation and metastasis by binding miR-128-3p and UPF1. Breast Cancer Res 2021;23:1-13. google scholar
- Dong Q, Cai N, Tao T, Zhang R, Yan W, Li R, et al. An axis involving SNAI1, microRNA-128 and SP1 modulates glioma progression. PLoS One 2014;9(6):e98651. google scholar
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2- AACT method. Methods 2001;25(4):402-8. google scholar
- Salmena L, Poliseno L, Tay Y, Kats L, Pandolfi PP. A ceRNA hypothesis: the Rosetta Stone of a hidden RNA language? Celi 2011;146(3):353-8. google scholar
- Tan W, Liu B, Qu S, Liang G, Luo W, Gong C. MicroRNAs and cancer: Key paradigms in molecular therapy. Oncol Lett 2018;15(3):2735-42. google scholar
- Khan AP, Poisson LM, Bhat VB, Fermin D, Zhao R, Kalyana-Sundaram S, et al. Quantitative proteomic profiling of prostate cancer reveals a role for miR-128 in prostate cancer. Mol Cell Proteomics 2010;9(2):298-312. google scholar
- Huang CY, Huang XP, Zhu JY, Chen ZG, Li XJ, Zhang XH, et al. miR-128-3p suppresses hepatocellular carcinoma proliferation by regulating PIK3R1 and is correlated with the prognosis of HCC patients. Oncol Rep 2015;33(6):2889-98. google scholar
- Du X, Li Y, Lian B, Yin X. microRNA-128-3p inhibits proliferation and accelerates apoptosis of gastric cancer cells via inhibition of TUFT1. World J Surg Oncol 2023;21(1):47. google scholar
- Chang Y, Zhang Q, Dong Z, Gao P, Hai Y. MicroRNA-128 inhibits the development of human colon cancer by targeting Rho family GTPase 3. J Taibah Univ Med Sci 2022;16(1):139-46. google scholar
- Pan J, Zhou C, Zhao X, He J, Tian H, Shen W, et al. A two-miRNA signature (miR-33a-5p and miR-128-3p) in whole blood as potential biomarker for early diagnosis of lung cancer. Sci Rep 2018;8(1):16699. google scholar
- Yang YR, Zang SZ, Zhong CL, Li YX, Zhao SS, Feng XJ. Increased expression of the lncRNA PVT1 promotes tumorigenesis in non-small cell lung cancer. Int J Clin Exp Pathol 2014;7(10):6929. google scholar
- Li X, Ren H. Long noncoding RNA PVT1 promotes tumor cell proliferation, invasion, migration and inhibits apoptosis in oral squamous cell carcinoma by regulating miR-150-5p/GLUT-1. Oncol Rep 2020;44(4):1524-38. google scholar
- Cui D, Yu CH, Liu M, Xia QQ, Zhang YF, Jiang WL. Long non-coding RNA PVT1 as a novel biomarker for diagnosis and prognosis of non-small cell lung cancer. Tumor Biol 2016;37:4127-34. google scholar
- Jin L, Cai Q, Wang S, Wang S, Wang J, Quan Z. Long noncoding RNA PVT1 promoted gallbladder cancer proliferation by epigenetically suppressing miR-18b-5p via DNA methylation. Cell Death Dis 2020;11(10):871. google scholar
- Wu D, Li Y, Zhang H, Hu X. Knockdown of Lncrna PVT1 enhances radiosensitivity in non-small cell lung cancer by sponging Mir-195. Cell Physiol Biochem 2017;42(6):2453-66. google scholar
- Zhao L, Kong H, Sun H, Chen Z, Chen B, Zhou M. LncRNA-PVTI promotes pancreatic cancer cells proliferation and migration through acting as a molecular sponge to regulate miR-448. J Cell Physiol 2018;233(5):4044-55. google scholar
- Zeng SG, Xie JH, Zeng QY, Dai SH, Wang Y, Wan XM, et al. Long noncoding RNA PVT1 promotes metastasis of non-small cell lung cancer through EZH2-mediated activation of hippo/NOTCH1 signaling pathways. Cell J 2021;23(1):21-31. google scholar
- Hu J, Gao W. Long noncoding RNA PVT1 promotes tumour progression via the miR-128/ZEB1 axis and predicts poor prognosis in esophageal cancer. Clin Res Hepatol Gastroenterol 2021;45(4):101701. google scholar