The Expression Of CXCR2 And MHC-? In Lung Adenocarcinoma

Objective:MHC-? represents an important part of adaptive immunity in antigen presenting cells;however,recent studies have suggested that it is also expressed on the surface of AT-? cells,which modulate the immune responses in the lung.As non-professional antigen presenting cells,AT-? cells express high levels of MHC-?,but lack CD80 and CD86 co-stimulatory molecules,and so are unable to activate Th1 CD4~+T cells.Although presentation of antigens by MHC-? provides the important‘signal one'for CD4 activation,co-stimulatory molecules CD80 and CD86,which provide‘signal two',are also necessary.The presentation of antigen by MHC-? to T cells in the absence of co-stimulatory molecules is a hallmark of the induction of anergy,a form of tolerance.Since AT-? cells do not express co-stimulatory molecules,antigen presentation by MHC-? expressed in AT-? cells is insufficient to activate effective CD4 T cells,but induces T-cell tolerance.It has been shown that the upregulation of MHC-?,as well as the absence of CD80 and CD86,in human AT-? cells is associated with Tregs expansion.In the urethane-induced TNF-?-mediated inflammation-driven lung adenocarcinoma(IDLA)model,we found that the MHC-? upregulation in lung adenocarcinoma,which arises from AT-? cells,was also associated with increased Foxp3 expression,as well as CD4~+CD25~+T cell infiltrations.When the AT-? cells isolated from lung adenocarcinoma tissues were co-cultured with spleen CD4~+T cells,we found that the MHC-? played a key role in Treg expansion.However,the mechanism about TNF-?-mediated inflammation upregulating MHC-? on AT-?-originated lung adenocarcinoma cells is still unknown.Recently,Kuang-Hui Sun demonstrated that TNF-a,a key mediator in the inflammatory tumour microenvironment,strongly up-regulated CXCR2expression to enhance migration,invasion,EMT and sphere formation of RCC cells.Yang showed that CXCR2 via NF-?B signaling pathways played a critical role in ovarian cancer progression by regulating the cell cycle,apoptosis,and angiogenesis.The function of CXCR2 and its ligands has been linked to cell survival in association with the NF-?B signaling network.CXCR2 deficiency decreased NF-?B expression and protected from cigarette smoke(CS)-induced lung inflammatory responses.Since CXCR-2 may affect NF-?B signaling pathway,it is necessary to explore the role of CXCR-2 in MHC-? expression in AT-? cells upon TNF-a-induced lung inflammation.Since we found TNF-?promotes upregulation of human MHC-? molecule(HLA-DR)in human AT-? cells in vitro,it is necessary to explore whether CXCR-2 regulates MHC-? expression in AT-?-originated lung adenocarcinoma cells in inflammation-driven lung adenocarcinoma.To address this question,it is important to analyze the correlation between MHC-? and CXCR-2 in lung adenocarcinoma.First,we applied the urethane to induce IDLA in mice,and explored the expression of MHC-? and CXCR-2 on AT-?-originated lung adenocarcinoma cells.Then,we collected human lung adenocarcinoma samples and explored whether CXCR-2 is correlated with HLA-DR expression in lung adenocarcinoma which arises from AT-? cells.Method:1.We used urethane to induce lung inflammation(8 weeks)as well as adenocarcinoma(6 months)in mice.After mice were subjected to urethane-treatment for 8 weeks,chronic lung inflammatory responses were induced in their lung tissues;When the mice were treated with urethane for 8 weeks,and then followed for a further 4 months alive,the mice developed lung adenocarcinoma.The expression of MHC-? and CXCR-2 was measured by immunohistochemical staining;colocilization of MHC-? and CXCR-2 on inflamed lung tissues as well as lung adenocarcinoma was measured by immunofluorescence analysis.2.Human paraffin-embedded cancer samples were obtained from the Department of Pathology,Hebei Chest Hospital,from 2010 to 2015,with the Institutional Internal Review Board's approval.A total of 44 cases of lung adenocarcinoma(LA),and 20 cases of normal lung tissue(NLT)were collected in the study.Patients who received neoadjuvant,or any type of postoperative therapy,or had another malignancy,were excluded.The pathologic diagnoses were established and reconfirmed according to the criteria of the 2015 World Health Organization(WHO)classification,based on morphologic,immunohistochemistry,cytogenetics,and molecular findings.Diagnostic slides from all cases were reviewed and agreed by two pathologists.The 44 cases of LA were further confirmed by positive immunohistochemical staining of TTF-1,and histopathologic data and the overall survival information of the patients is available.The expression of HLA-DR and CXCR-2inhumanlungadenocarcinomawasmeasuredby immunohistochemical staining and immunofluorescence analysis.Then,the correlation of HLA-DR and CXCR-2 was measured.Result:1.The increased expression of MHC-? and CXCR-2 in urethane-induced,inflammation-driven,lung adenocarcinoma.After urethane-treatment for 8weeks,chronic lung inflammatory responses were induced in mice lung tissues.When the mice were treated with urethane for 8 weeks and then keep alive for another 4 months,lung adenocarcinoma was induced.Immunohistochemical results showed that all urethane-induced lung adenocarcinoma exhibited AT-? cells specific marker SP-C expression.MHC-? was mostly localized on thyroid transcription factor-1(TTF-1+)AT-? cells,and the number of MHC-?+TTF-1+cells was increased in the inflamed lung tissues.The results confirmed that urethane could induce inflammation-driven lung adenocarcinoma,which arises from AT-? cells.In inflamed lung tissues and lung adenocarcinoma,immunohistochemical staining showed increased expression of CXCR-2,higher than that in control lung tissues,which was associated with increased MHC-? expression.Furthermore,we found CXCR-2 and MHC-? colocalization by immuofluorescence staining,and CXCR-2+MHC-?+positive cells were increased in urethane-induced inflamed lung tissues and lung adenocarcinoma,which suggests that CXCR-2 may be associated with MHC-? expression in inflammation-driven lung adenocarcinoma.2.The expression of HLA-DR and CXCR-2 in human lung adenocarcinoma samplesTo further validate our findings in the lung adenocarcinoma mouse model,we analyzed the correlations between CXCR-2 and HLA-DR in human lung adenocarcinoma.Immunohistochemical staining showed that positive expression of HLA-DR and CXCR-2 was shown in human lung adenocarcinoma.Higher expression of HLA-DR was observed in lung adenocarcinoma compared with those in control tissues.Furthermore,we found that high expression of CXCR-2 was also correlated with TNM stages.3.The correlation between MHC-? and CXCR-2 expression in human lung cancer specimens.Double-color immuofluorescence analysis showed that CXCR-2 and MHC-? colocalization was detected in lung adenocarcinoma.There is a positive correlation between CXCR-2 and HLA-DR expression in lung adenocarcinoma.Thus,those findings suggest that CXCR-2 may be associated with MHC-? expression in AT-? cells in inflammation-driven lung adenocarcinoma.Conclusion:1.In urethane-induced,inflammation-driven,lung adenocarcinoma mice model,co-expression of MHC-? and CXCR-2 was observed on AT-?-originated lung adenocarcinoma cells,which suggests it is possible that CXCR-2 may be related with MHC-? expression in IDLA.2.High expression of CXCR-2 and HLA-DR was observed in human lung adenocarcinoma samples.CXCR-2 and MHC-? colocalization was detected in lung adenocarcinoma,and there is a positive correlation between CXCR-2 and HLA-DR expression in lung adenocarcinoma of AT-? cells origin.Thus,those findings suggest that CXCR-2 may be associated with MHC-? expression in AT-? cells in inflammation-driven lung adenocarcinoma.