ObjectiveTo explore and analyze the distribution of NAT2 gene in Han population in Shenzhen andprovidethebasisfortreatmentof NAT2-relatedmetabolicdiseasesandcancer research. MethodThe study diluted the DNA sample(d0 ng/μl) as follows: 1 × 100, 1 × 101, 1 × 102, 1 ×103 , 1 × 104, to verify the sensitivity of detection of NAT2 gene polymorphism by real-time PCR. Mutation locus of 282,341,481, 590 and 857 of NAT2 in 554 normal samples were detected and genotyped by realtime PCR with Taqman probes. Forty-seven cases among 554 healthy controls were detected by DNA sequencing to verify the sensitivity, specificity and accuracy of this assay. ResultsThe lowest detection limit was 10-4 ng/μl. The frequencies of each allele of NAT2 were: *4 allele 64. 6％ (358/554), *5 allele 6. 3％ (35/554), *6 allele 25. 3％ ( 140/554), *7 allele 30. 0％ ( 166/554), * 11 allele 0. 6％ (3/554) The frequencies of main genotypes of NAT2 were as the follows : NAT2 *4/* 6 12.3％ ( 68/554 ), *6/* 78. 3％ (46/554), * 13/* 13 or * 12/* 12 or *4/* 4 7.6％ (42/554), *4/* 7 8. 1％ (45/554), * 7/* 1 312. 3％ (68/554), * 12 7.2％ (40/554), *6/* 13( * 12)5.6％ (31/554). The samples with 7 genotypes account for 89. 6％ (496/554) of the total cases.Rapid acetylation and Intermediate type accounting for 40. 5％ (224/554) and 46. 7％ (259/554) respectively were the main phenotypes of NAT2 in Han people in Shenzhen. In addition, the accuracy, specificity and sensitivity of the PCR were compared to direct DNA sequencing. The accuracy of 282, 341,481, 590 and 857 were 88.2％ (30/34), 87.5％ (7/8), 80. 0％(4/5), 100. 0％ ( 22/22 ) and 93.8％ ( 15/16 ) respectively. The specificities were 100. 0％ ( 13/13 ),94. 9％ ( 37/39), 100. 0％ ( 42/42 ) , 96.0％ ( 24/25 ) and 96. 8％ ( 30/31 ) respectively. The sensitivities were 91.5％ ( 43/47 ), 93.6％ ( 44/47 ), 97.6％ ( 46/47 ) 97.9％ ( 46/47 ) and 95.7％ ( 45/47 ),respectively. ConclusionsThe sensitivity, specificity and accuracy of real-time PCR established in this study is good. It can be widely used for clinical and scientific research. The major alleles of NAT2 of Han population in Shenzhen are * 4, *6, * 7. The most common slow acetylation genotype is *6/* 7. This study can provide a basis for NAT2-related metabolic diseases and cancer research.

BACKGROUND: In China, lung cancer remains the cancer with the highest incidence and mortality rate. Among early-stage lung adenocarcinomas (LUAD), the micropapillary (MPP) component is prevalent and typically exhibits high aggressiveness, significantly correlating with early metastasis, lymphatic infiltration, and reduced five-year survival rates. Therefore, the study is to explore the similarities and differences between MPP and non-micropapillary (non-MPP) components in malignant pulmonary nodules characterized by GGOs in early-stage LUAD, identify unique mutational features of the MPP component and analyze the relationship between the ZNF469 gene, a member of the zinc-finger protein family, and the prognosis of early-stage LUAD, as well as its correlation with immune infiltration. METHODS: A total of 31 malignant pulmonary nodules of LUAD were collected and dissected into paired MPP and non-MPP components using microdissection. Whole-exome sequencing (WES) was performed on the components of early-stage malignant pulmonary nodules. Mutational signatures analysis was conducted using R packages such as maftools, Nonnegative Matrix Factorization (NMF), and Sigminer to unveil the genomic mutational characteristics unique to MPP components in invasive LUAD compared to other tumor tissues. Furthermore, we explored the expression of the ZNF469 gene in LUAD using The Cancer Genome Atlas (TCGA) database to investigate its potential association with the prognosis. We also investigated gene interaction networks and signaling pathways related to ZNF469 in LUAD using the GeneMANIA database and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Lastly, we analyzed the correlation between ZNF469 gene expression and levels of immune cell infiltration in LUAD using the TIMER and TISIDB databases. RESULTS: MPP components exhibited a higher number of genomic variations, particularly the 13th COSMIC (Catalogue of Somatic Mutations in Cancer) mutational signature characterized by the activity of the cytidine deaminase APOBEC family, which was unique to MPP components compared to non-MPP components in tumor tissues. This suggests the potential involvement of APOBEC in the progression of MPP components in early-stage LUAD. Additionally, MPP samples with high similarity to APOBEC signature displayed a higher tumor mutational burden (TMB), indicating that these patients may be more likely to benefit from immunotherapy. The expression of ZNF469 was significantly upregulated in LUAD compared to normal tissue, and was associated with poor prognosis in LUAD patients (P<0.05). Gene interaction network analysis and GO/KEGG enrichment analysis revealed that COL6A1, COL1A1, COL1A2, TGFB2, MMP2, COL8A2 and C2CD4C interacted with ZNF469 and were mainly involved in encoding collagen proteins and participating in the constitution of extracellular matrix. ZNF469 expression was positively correlated with immune cell infiltration in LUAD (P<0.05). CONCLUSIONS The study has unveiled distinctive mutational signatures in the MPP components of early-stage invasive LUAD in the Asian population. Furthermore, we have identified that the elevated expression of mutated ZNF469 impacts the prognosis and immune infiltration in LUAD, suggesting its potential as a diagnostic and prognostic biomarker in LUAD.
Humans ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung/genetics* ; China ; Prognosis ; Transcription Factors