Development of standard templates for absolute quantification of some long non-coding rnas for hepatocellular carcinoma research

  • Bùi Lan Anh Học viện Quân y
  • Lê Thị Hồng Vân Bệnh viện Quân y 103
  • Nguyễn Văn Tâm Bệnh viện Quân y
  • Phạm Nhật Hằng Trường ĐHKHTN, Đại học Quốc gia Hà Nội
  • Hoàng Văn Tổng Học viện Quân y

Main Article Content

Keywords

Long non-coding RNAs (lncRNAs), cloning, standard templates.

Abstract

Objective: Hepatocellular carcinoma (HCC) is one of the cancers with the highest mortality rate. Long non-coding RNAs (lncRNAs), a class of non-protein-coding RNAs, have been recognized as potential biomarkers for early cancer diagnosis. This study aims to develop a lncRNA panel samples for absolute quantification of lncRNAs. Subjects and Methods: Five lncRNAs (LINC02560, LINC02487, HCG22, HNF4A-AS1, and C5orf66-AS1) were selected. Recombinant cloning was performed using PCR amplification, DNA recombination, and propagation in E. coli. Standard DNA templates were developed, and absolute quantification was conducted using real-time PCR. Results: Recombinant vectors harboring sequences of the five target lncRNAs were successfully cloned into E. coli. Standardized reference materials for absolute quantification were established, demonstrating high amplification efficiency (95.45 - 109.14%), strong specificity, good reproducibility (CV% < 10%), and high linearity (R2 > 0.99). Conclusion: The results provide standard panel samples that enable the accurate identification of lncRNA biomarkers in HCC research, supporting early diagnosis and prognostic evaluation in liver cancer.

Article Details

References

1. Bray F, Laversanne M, Sung H et al (2024) Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. 74(3): 229-263. doi:https://doi.org/10.3322/caac.21834
2. Observatory GC (2025) GLOBOCAN 2022: Viet Nam Fact Sheet. International Agency for Research on Cancer (IARC), World Health Organization. Updated https://gco.iarc.who.int/media/globocan/factsheets/populations/704-viet-nam-fact-sheet.pdf
3. Patresan J, Patel H, Chandrasekaran K, Reynolds G (2024) Current Treatment Paradigm and Approach to Advanced Hepatocellular Carcinoma. Cureus 16(12): 75471. doi:10.7759/cureus.75471
4. Mittal S, El-Serag HB (2013) Epidemiology of hepatocellular carcinoma: consider the population. J Clin Gastroenterol 47 Suppl(0): 2-6. doi:10.1097/MCG.0b013e3182872f29
5. Li Y, Egranov SD, Yang L, Lin C (2019) Molecular mechanisms of long noncoding RNAs-mediated cancer metastasis. Genes Chromosomes Cancer 58(4): 200-207. doi:10.1002/gcc.22691
6. Jansone B, Kadish I, van Groen T et al (2016) Memory-enhancing and brain protein expression-stimulating effects of novel calcium antagonist in Alzheimer’s disease transgenic female mice. Pharmacological Research 113: 781-787. doi:https://doi.org/10.1016/j.phrs.2016.06.020
7. Sang J, Wang Z, Li M et al (2017) ICG: A wiki-driven knowledgebase of internal control genes for RT-qPCR normalization. Nucleic Acids Research. 46(D1):D121-D126. doi:10.1093/nar/gkx875
8. Hays A, Islam R, Matys K, Williams D (2022) Best Practices in qPCR and dPCR Validation in Regulated Bioanalytical Laboratories. The AAPS Journal 24(2): 36. doi:10.1208/s12248-022-00686-1.
9. Hashish A, Sinha A, Sato Y, Macedo NR, El-Gazzar M (2022) Development and Validation of a New TaqMan Real-Time PCR for the Detection of Ornithobacterium rhinotracheale. Microorganisms 10(2): 341.
10. Li X, Xiao X, Chang R, Zhang C (2020) Comprehensive bioinformatics analysis identifies lncRNA HCG22 as a migration inhibitor in esophageal squamous cell carcinoma. Journal of Cellular Biochemistry 121(1): 468-481. doi:https://doi.org/10.1002/jcb.29218
11. Wapinski O, Chang HY (2011) Long noncoding RNAs and human disease. Trends in Cell Biology 21(6): 354-361. doi:10.1016/j.tcb.2011.04.001
12. Quinn JJ, Chang HY (2016) Unique features of long non-coding RNA biogenesis and function. Nature Reviews Genetics 17(1): 47-62. doi:10.1038/nrg.2015.10
13. Tang X, Shang J, Ji Y, Sun Y (2023) PLASMe: A tool to identify PLASMid contigs from short-read assemblies using transformer. Nucleic Acids Res 51(15): 83. doi:10.1093/nar/gkad578
14. Bustin SA, Benes V, Garson JA et al (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. Apr 55(4): 611-22. doi:10.1373/clinchem.2008.112797
15. Nguyễn MN, Nguyễn TTD, Đinh TTH và cộng sự (2024) Xây dựng đường chuẩn để định lượng nồng độ long non-coding RNA IFI6lnc2 và EPB41L4A-AS1 bằng phương pháp Real-time PCR. Tạp chí Y Dược học Quân sự 49(2): 245-255. doi:10.56535/jmpm.v49i2.720

Article Sidebar

PDF (Tiếng Việt)
Published
May 15, 2026
DOI: https://doi.org/10.52389/ydls.v21i4.3035
Issue
Volume 21 - No 4/2026
Section
Articles
How to Cite
AnhB. L., Hồng VânL. T., TâmN. V., HằngP. N., & TổngH. V. (2026). Development of standard templates for absolute quantification of some long non-coding rnas for hepatocellular carcinoma research. Tạp Chí Y Dược lâm sàng 108, 21(4). https://doi.org/10.52389/ydls.v21i4.3035