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Published: 2025-07-28
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DOI: https://doi.org/10.52852/tcncyh.v192i7.3598
Issue
Vol. 192 No. 7 (2025)
Section
Các bài báo
How to Cite
Mậu, N. T. T., Đạt, Đào X., Khánh, T. V., Thịnh, T. H., Dung, H. M., Hằng, Đỗ T. L., & Bình, N. T. (2025). 8. Development of a procedure to evaluate mRNA expression of the ifnγ gene in cultured gamma delta T cells from lung cancer patients utilizing real-time PCR method. Journal of Medical Research, 192(7), 65-73. https://doi.org/10.52852/tcncyh.v192i7.3598

8. Development of a procedure to evaluate mRNA expression of the ifnγ gene in cultured gamma delta T cells from lung cancer patients utilizing real-time PCR method

Nguyen Thi Thuy Mau, Dao Xuan Dat, Tran Van Khanh, Tran Huy Thinh, Ho My Dung, Do Thi Le Hang, Nguyen Thanh Binh

Main Article Content

Abstract

This study aimed to develop a procedure for evaluating the mRNA expression of the IFNγ gene in cultured γδT cells derived from lung cancer patients using Real-time PCR technique. This serves as a preliminary step to support the role of γδT cells and the IFNγ cytokine in autologous immune cell therapy for lung cancer in Vietnam. The study was conducted on 10 peripheral blood samples collected from patients with non-small cell lung cancer. γδT cells were isolated and cultured for 14 days. Total RNA was then extracted, reverse-transcribed into cDNA, and analyzed using semi-quantitative Real-time PCR to assess IFNγ gene expression. Gene expression was compared between day 0 and day 14 of culture. The results showed a significant increase in IFNγ mRNA expression after 14 days of γδT cell culture, with an average fold change of 3.21 ± 1.42 compared to pre-culture levels. The optimal annealing temperature was determined to be 550C, and the most effective primer concentrations were 14.2 pmol/μl for β-actin and 12 pmol/μl for IFNγ, yielding clear, specific bands and consistent expression data. A reliable procedure for evaluating IFNγ mRNA expression in cultured γδT cells using Real-time PCR was successfully established.

Article Details

Keywords

γδT cells, cytokine IFNγ, lung cancer, gene expression.

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