Objective:To discuss the expressions of autotaxin(ATX)and lysophosphatidic acid receptor 3(LPA3)in serum and lung tissue of the chronic obstructive pulmonary disease(COPD)patients,and to clarify the role of ATX and LPA3 in the occurrence and development of COPD.Methods:A total of 40 COPD patients were collected and brought into acute exacerbation of COPD group(AECOPD group);after treatment,those stabilized patients were included in COPD stable group(n=40);additionally,40 healthy individuals were recruited as control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the ATX levels in the serum of the subjects in various groups.Another 80 patients who underwent lobectomy were divided into COPD smoking group(CS group,n=20),COPD non-smoking group(CNS group,n=20),non-COPD smoking group(HS group,n=20),and non-COPD non-smoking group(HNS group,n=20).The general informations of the subjects in various groups were collected.HE staining was used to observe the pathomorphology of lung tissue of the patients in various groups;immunohistochemical staining was used to detect the expression levels of ATX and LPA3 proteins in lung tissue of the patients in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of ATX and LPA3 mRNA in lung tissue of the patients in various groups;Pearson correlation analysis was used to evaluate the correlations of continuous variables with normal distribution,and Spearman correlation analysis was used to evaluate the correlations of other variables.Results:Compared with control group,the percentage of forced expiratory volume in the first second to predicted value(FEV1％pred)and the percentage of forced expiratory volume in the first second to forced vital capacity(FEV1/FVC)of the patients in COPD stable group were significantly decreased(P＜0.05).Compared with COPD stable group and control group,the level of ATX in serum of the patients in AECOPD group was increased(P＜0.05);compared with control group,the level of ATX in serum of the patients in COPD stable group was increased(P＜0.05).The serum ATX level of the patients in AECOPD group was positively correlated with COPD Assessment Test(CAT)scores(r=0.581,P＜0.001)and showed no correlations with smoking history,white blood cells(WBC),percentage of neutrophils(NEUT％),neutrophil to lymphocyte ratio(NLR),and body mass index(BMI)(P＞0.05).The serum ATX level of the patients in COPD stable group was positively correlated with WBC and CAT score(r=0.384,P=0.014;r=0.463,P=0.003)and negatively correlated with FEV1％pred and FEV1/FVC(r=-0.393,P=0.012;r=-0.353,P=0.025).Compared with CS group and CNS group,the FEV1％pred and FEV1/FVC of the patients in HS group and HNS group were increased(P＜0.05).The immunohistochemical staining results showed that compared with CS group,the expression levels of ATX and LPA3 proteins in lung tissue of the patients in HS group and HNS group were decreased(P＜0.05).Compared with CNS group,the expression levels of ATX and LPA3 proteins in lung tissue of the patients in HS group and HNS group were decreased(P＜0.05).The RT-qPCR results showed that compared with CS group,the expression levels of ATX and LPA3 mRNAs in lung tissue of the patients in HS group and HNS group were decreased(P＜0.05);compared with CNS group,the expression levels of ATX and LPA3 mRNAs in lung tissue of the patients in HS group and HNS group were decreased(P＜0.05).The correlation analysis results showed that the expression level of ATX protein in lung tissue of the COPD patients was positively correlated with the expression level of LPA3 protein(r=0.723,P＜0.001).Conclusion:The expression levels of ATX and LPA3 mRNA and proteins in lung tissue of the COPD patients are increased.The serum ATX levels in both AECOPD group and COPD stable group are increased,suggesting that these factors may be involved in the inflammatory response of COPD,promoting its occurrence and development.

Humans ; COVID-19 ; SARS-CoV-2

BACKGROUNDNon-small cell lung cancer (NSCLC) is the most common lung malignancy worldwide. The metastatic potential of NSCLC cells has been shown to be associated with the tumor microenvironment, which consists of tumor cells, stroma, blood vessels, immune infiltrates and the extracellular matrix. Fibroblasts can produce numerous extracellular matrix molecules and growth factors. Gefitinib has been evaluated as a first-line treatment in selected patients, and it has shown favorable efficacy especially in NSCLC, but it is not effective for everyone.

METHODSIn this study, we examined the antitumor activity of gefitinib on lung fibroblasts co-cultured of lung cancer cells. A series of co-culture experiments that employed cell counting kit-8 (CCK8), transwells, real-time polymerase chain reaction (RT-PCR) and Western blotting with HFL-1 fibroblasts and A549 human lung carcinoma cells were performed to learn more about tumor cell proliferation, migration and invasion; and to determine any change of epithelial mesenchymal transition (EMT)-associated tumor markers vimentin, matrix metallopro-teinase 2 (MMP2) and chemotaxis cytokines receptor 4 (CXCR4) mRNA levels.

RESULTSA549 cell proliferation in the presence of HFL-1 cells was not significantly increased compared with A549 cells alone, but A549 cell spheroid body formation was increased after co-culture, and treatment with gefitinib increased further. Our study also revealed that fibroblasts attenuated the lung cancer cell inhibition ratio of migration and invasion after gefitinib treatment in vitro. To further study this mechanism, RT-PCR analysis showed that vimentin, MMP2 and CXCR4 mRNA levels were more highly expressed in the lung cancer cells after co-culture, but did not obviously decrease compared with the control cells following gefitinib treatment. This suggests the mechanism by which fibroblasts attenuate gefitinib-induced expression of EMT-associated tumor markers. Finally, our results demonstrated that co-culture with A549 lung cancer cells does not alter the cell cycle distribution of HFL-1 fibroblasts. Furthermore, HFL-1 fibroblasts had no effect on the cell cycle distribution of HFL-1 cells treated with gefitinib.

CONCLUSIONGefitinib has lower anti-tumor activity on A549 lung cancer cells when co-cultured with HFL-1 fibroblasts.

Antineoplastic Agents ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Coculture Techniques ; Epithelial-Mesenchymal Transition ; drug effects ; Fibroblasts ; cytology ; drug effects ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Quinazolines ; pharmacology