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Published: 2025-07-28
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DOI: https://doi.org/10.52852/tcncyh.v192i7.3601
Issue
Vol. 192 No. 7 (2025)
Section
Các bài báo
How to Cite
Mậu, N. T. T., Thịnh, T. H., Đạt, Đào X., Hằng, Đỗ T. L., & Bình, N. T. (2025). 9. Optimization of the evaluation protocol for TNFα gene expression in cultured natural killer cells from lung cancer patients using real-time PCR method. Journal of Medical Research, 192(7), 74-82. https://doi.org/10.52852/tcncyh.v192i7.3601

9. Optimization of the evaluation protocol for TNFα gene expression in cultured natural killer cells from lung cancer patients using real-time PCR method

Nguyen Thi Thuy Mau, Tran Huy Thinh, Dao Xuan Dat, Do Thi Le Hang, Nguyen Thanh Binh

Main Article Content

Abstract

This study aimed to establish and optimize a standardized protocol for evaluating TNFα gene expression in natural killer (NK) cells cultured from peripheral blood of patients with non- small cell lung cancer (NSCLC). Peripheral blood samples were collected from 10 patients diagnosed with NSCLC, NK cells were isolated and expanded under controlled culture conditions to enhance cellular activation and proliferation. The protocol was optimized with respect to critical PCR and Real-time PCR parameters, including annealing temperature, primer concentration, and the use of β-actin as an internal control gene. TNFα expression levels were quantified using the 2-∆∆Ct method. After 21 days of culture, TNFα expression showed a mean increase of 2.22 ± 1.08- fold compared to baseline, while β-actin expression remained stable across time points, ensuring the reliability of quantitative analyses. The optimized protocol provides an effective approach for quantifying TNFα gene expression in NK cells, serving as an indicator of enhanced immune functionality. These findings support the potential application of ex vivo-expanded NK cells as an adjunctive strategy in cancer immunotherapy, particularly within the clinical context of Vietnam.

Article Details

Keywords

TNFα, NK cells, Real-time PCR, lung cancer, gene expression

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