原发性肺腺癌患者外周血端粒酶逆转录酶阳性白细胞检测方法的建立

doi:10.3969/j.issn.1004-616x.2024.06.002

Abstract

Abstract: OBJECTIVE： This study aimed to develop a method for detecting white blood cell subsets with high telomerase reverse transcriptase (TERT) activity to evaluate proliferative potential of white blood cells in patients with malignant tumors. METHODS： A modified recombinant type I herpes simplex virus (HSV1-hTERTp-eGFP) was able to replicate in white blood cells with high TERT promoter transcriptional activity and to express green fluorescent protein (GFP). By observing green fluorescence，TERT positive white blood cells with high proliferation potential was identified，and the immune proliferation potential of individuals was evaluated. The addition of 100，500 and 1 000 GFP positive Jurkat cells that were infected with HSV1-hTERTp-eGFP virus for 24 hours to 2.5×10⁶ peripheral blood white blood cells that have not been infected with the virus were used as simulated samples for detection. Three parallel groups were set up and were tested for recovery rate，repeatability，linear regression coefficient，and other indicators. At the same time，peripheral blood samples were collected from 5 patients with benign pulmonary nodules，and 3 parallel groups were set up to detect the proportion of TERT positive white blood cells in 1 mL of peripheral blood in each group. The technical reliability of detecting peripheral blood samples was evaluated by calculating the coefficient of variation within each group. Using this detection technology，the percentage of TERT positive white blood cells in peripheral blood of 80 patients with benign pulmonary nodules and 80 patients with primary lung adenocarcinoma were detected and analyzed. RESULTS： In the simulated samples with Jurkat cells，the average number of positive cells were 80.3，448.2 and 920.6，respectively，with recovery rates of 80.3%，89.6% and 92.1%，respectively；The inter batch coefficients of variation for the three repeated experiments were 11.3%，7.2% and 4.4%，respectively，while the intra batch coefficients of variation within the three parallel experimental groups were less than 10%；The intra group coefficient of variation for the percentage of TERT positive cells in peripheral blood of 5 patients with benign pulmonary nodules was also within 10%. This detection technology showed high accuracy and stability. In 80 patients with benign pulmonary nodules，the percentage of TERT positive white blood cells showed a decreasing trend with age (r=-0.61；P<0.01). Compared with the control group of benign pulmonary nodule patients，the percentage of TERT positive white blood cells in early lung adenocarcinoma patients decreased (P<0.01)；Compared with non-invasive lung adenocarcinoma patients，the proportion of TERT positive white blood cells in invasive lung adenocarcinoma patients was significantly reduced (P<0.01). CONCLUSION： The peripheral blood immune cells of patients with lung adenocarcinoma showed a decrease in proliferation ability，which was more pronounced in patients with invasive lung adenocarcinoma. The observation was reminiscent to immunosenescence，and would provide a better understanding of poor prognosis among patients with invasive lung adenocarcinoma.

Key words: lung adenocarcinoma, telomeres, telomerase reverse transcriptase, immunosenescence

CLC Number:

R734.2

XIE Peipei, ZHANG Qi, ZHANG Xiaoli, WAN Duo, ZHANG Wen, CHENG Shujun, ZHANG Kaitai. Development of a detection method for TERT positive white blood cells in patients with primary lung adenocarcinoma[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(6): 431-437,443.

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Development of a detection method for TERT positive white blood cells in patients with primary lung adenocarcinoma

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