| Objective:To determine the connection between the expression of the inhibitory receptor LAIR-1(Leukocyte-associated Immunoglobulin-like Receptor-1)levels with the clinical pathological characteristics of NSCLC(Non-small Cell Lung Cancer),and its prognostic implication.Materials and methods:Download and extract TCGA(The Cancer Genome Atlas)database and GEO(Gene Expression Omnibus database)LAIR-1 m RNA and clinical data from patients with NSCLC.Applying the Wilcoxon test to the analysis of LAIR-1 m RNA expression differences in different tissues,and using the Kaplan-Meier method and Log-rank test to plot the survival curve.Tissue samples from NSCLC patients and paired adjacent cancer tissues were collected and the expression of LAIR-1 in NSCLC tissues detected the relationship between LAIR-1 gene expression and prognosis was validated by immunohistochemistry.A correlation study between LAIR-1 gene expression and clinicopathological characteristics was analyzed by chi-square test,and multifactorial analysis was carried out using logistic regression analysis.Application of single factor and multi-factor COX analysis of factors effect of survival in patients with NSCLS.LAIR-1 expression in human bronchial epithelial cell line 16 HBE,NSCLC cell lines(A549,SK-MES-1)was verified in vitro by Real-time PCR assay and Western Blotting analysis.A549 cell lines and SK-MES-1 cell lines overexpressing LAIR-1 gene were transfected with lentivirus,and overexpression model establishment was verified by fluorescent staining,Real-time PCR assay,and Western Blotting analysis.Flow cytometry analysis of LAIR-1 gene effects on cell cycle of NSCLC cell lines after PI reagent staining.Flow cytometry was used to detect apoptosis following Annexin VAPC and 7-AAD staining,and the effect on apoptosis was analyzed by T-Test.Results:1.The m NA analysis of the TCGA database revealed that LAIR-1 was lowly expressed in NSCLC tissues and less than in adjacent tumor tissues(P < 0.01).In addition,the clinical data analysis of TCGA and GEO databases revealed OS was lower in the LAIR-1 low-expression group compared with the LAIR-1 high-expression group(P < 0.05).2.Immunohistochemistry data confirmed that LAIR-1 was less expressed in NSCLC tissues than in para-cancer tissues(P<0.05),among them,LAIR-1 was expression levels of the lowest in squamous tissues(P<0.05).Univariate analysis revealed that the low expression of LAIR-1 was related to multiple clinicopathological characteristics,such as lymph node metastasis(P=0.025),tumor stage(P=0.023),gender(P=0.003),pathological type(P=0.000)and smoking history(P=0.001),OS(P=0.000)and PFS(P=0.000)in NSCLC patients.Multivariate analyses demonstrated that the expression of low LAIR-1 gene expression in squamous cell carcinoma patients(P=0.000)was more prone to NSCLC progression(P=0.046).3.OS and PFS were lower in the LAIR-1 low-expression group than in the LAIR-1high-expression group(P = 0.000).COX regression univariate analysis showed that LAIR-1 expression(P=0.000)was correlated with PFS and OS,and clinical stage was correlated with PFS(P=0.007)and OS(P=0.014).The multifactorial analysis demonstrated that LAIR-1 expression(P = 0.000)and clinical stage(P=0.027)were independent prognostic factors for PFS,but only LAIR-1 expression(P = 0.000)was an independent prognostic factor for NSCLC OS.4.Validation of LAIR-1 expression was lower in the in vitro cell lines A549 and SKMES-1 cell line compared to 16 HBE cells,and LAIR-1 expression was lowest in the LUSC cell line SK-MES-1 cells.A549 cell lines and SK-MES-1 cell lines LAIR-1 gene overexpression models were successfully constructed.LAIR-1 overexpression blocked the SK-MES-1 cell lines in the G1 phase(P<0.05),did not significantly affect cells in the G2/M phase(P>0.05),and promoted apoptosis(P<0.05).In contrast,there was no effect on cell cycle and apoptosis in the A549 cell lines(P > 0.05).Conclusions:1.LAIR-1 was down-regulated in NSCLC,among them,the lowest expression was observed in squamous carcinoma.2.Patients with low LAIR-1 expression have a poorer prognosis and were more likely to progress.3.LAIR-1 expression status was also closely associated with gender,smoking history,tumor stage,pathological type,lymph node metastasis,OS,and PFS.Moreover,LAIR-1 promoted the progression of lung squamous carcinoma.Factors influencing OS and PFS in NSCLC are relevant to LAIR-1 expression,and clinical stage,with LAIR-1being an independent factor of PFS and OS in NSCLC.4.It was verified that LAIR-1 expression was lower in the in vitro cell lines A549 and SK-MES-1 cells compared to 16 HBE cells,and LAIR-1 expression was even lowest in SK-MES-1 cells.LAIR-1 overexpression blocks lung squamous carcinoma SK-MES-1 cells in the G1 phase and promotes apoptosis.In conclusion,LAIR-1performed an important role in the development of NSCL. |
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