The Effect Of RAGE And DIAPH1 On The Migration And Apoptosis Of Lung Adenocarcinoma Cell And Its Expression In Non-small Cell Lung Cancer

Background Lung cancer is one of the leading causes of morbidity and mortality in the world.It also becomes the leading cause of cancer death all over the world,which seriously endangers human health.Due to the capacity of invasion of the surrounding blood vessels in the early stages of lung cancer,further leads to a distant metastasis,resulting in the loss of surgical treatment.Therefore,it is very urgent to find out the effective therapeutic target to control the invasion and metastasis of lung cancer.Objects 1.To explore the expression of the receptor for advanced glycation end products(RAGE)and its intracellular signaling molecules DIAPH1 in lung adenocar-cinoma A549 cells and the impact on the ability of cell migration and apoptosis.2.To observe the expression level of RAGE and DIAPH1 in non-small cell lung cancer(NSCLC)to study its clinical significance.Methods 1.Using q-RT PCR,Western Bolt and immunocytofluorescence to test the expression and distribution of RAGE and DIAPH1 in lung adenocarcinoma A549 cells and human bronchial epithelial cells BEAS-2B.2.Treated lung adenocarcinoma A549 cells with different concentration(1μg、10μg、100μg)of RAGE ligands CML-AGE and S100 B,using wound healing test to identify the effect of migration ability.3.Treated lung adenocarcinoma A549 cells with different concentration(25μg、50μg、100μg)of RAGE ligands CML-AGE,using q-RT PCR to test the gene expression of BCL-2/BAX.4.Immunohistochemistry was used to determine RAGE and DIAPH1 protein expression in 30 NSCLC tissue samples and 10 cases of non-tumor adjacent tissues.Results 1.The results of q-RT PCR,Western Blot and immunocytofluorescence showed,compared with human bronchial epithelium cells BEAS-2B,the expression of RAGE and DIAPH1 were both significantly down-regulated in lung adenocarcinoma A549 cells(P<0.0001)2.After treated with RAGE ligands CML-AGE and S100 B in different concentration(1μg、10μg、100μg),16 hours later,compared with control group,the experimental group showed the ligands inhibit lung adenocarcinoma A549 cells migration in concentration-depend manners.(P<0.05)3.After treated with RAGE ligands CML-AGE in different concentration(25μg、50μg、100μg),compared with control group,the expression of anti-apoptotic gene BCL-2 was down-regulated and pro-apoptotic gene BAX was upregulated in the experimental group in concentration-depend manners.(P<0.05).4.Immunohistochemistry showed the expression level of RAGE was steady and high in non-tumor adjacent tissues and negative in NSCLC.Strong staining was observed in non-tumor adjacent tissue sections;weak staining was observed in NSCLC tissue sections.The expression of DIAPH1 was moderate to strong in both NSCLC and nontumor adjacent tissue sections.Conclusions1.The expression levels of RAGE and DIAPH1 in lung adenocarcinoma A549 cells were both significantly lower than human bronchial epithelium cells BEAS-2B.2.RAGE ligands CML-AGE and S100 B could inhibit lung adenocarcinoma A549 cells migration in concentration-depend manners.3.RAGE ligand CML-AGE could promote cell apoptosis in lung adenocarcinoma A549 cells in concentration-depend manners.4.The expression of RAGE in NSCLC was lower than non-tumor adjacent tissues;the expression of DIAPH1 in NSCLC and non-tumor adjacent tissues were both upregulated.