Expression And Significance Of STRAIL And TRAIL Receptors In Inhibitory On Pancreatic Cancer Cells By DNT Cells

Objective Our previous studies showed that the TCR+ CD3+ CD4- CD8- double negative(DNT) T cell could inhibit pancreatic cancer cells in vitro and vivo. To investigate the role and significance of the tumor necrosis Sfactor-related D apoptosis inducing ligands(s TRAIL) binding to its receptor TRAIL-R1 and TRAIL-R2 in DNT cell suppressing we employed this study.Methods DNT cells could be cultured according a novel of effective protocol of antibody adsorption, we collect the cells from the wells. ELISA methods were used to measure the levels of s TRAIL in the culture supernatants on day 12. Western blot, Real-time Quantitative Polymerase chain reaction(q PCR) were employed to compare the expression of TRAIL-R1 and TRAIL-R2 in 6 pancreatic cancer cell lines from molecular and protein levels. The expression of TRAIL-R1 and TRAIL-R2 in 48 cases of pancreatic ductal adenocarcinoma tissues and adjacent tissues who had been diagnosed as pancreatic cancer at Anhui Provincial Hospital could be observed by immunohistochemistry. we evaluate the association of TRAIL-R1 and TRAIL-R2 with clinical pathological parameters.Results The number of cells gradually was improved and DNT cells were cultured 12 days the total number of expanded DNT cells could reached peak. In DNT groups, the concentrations of s TRAIL were higher than in control groups(t=27.044, P<0.05). QPCR found that the relative quantities of DR4 mRNA were 0.807, 1, 0.423, 0.662, 1.470 and 1.418 in SW1990, PANC-1, BXPC-3, MIA Pa Ca-2, CFPAC-1 and Capan-1, respectively. Similarly, the relative quantities of DR5 in cell lines as follow, 0.744(SW1990), 1(PANC-1), 0.999(BXPC-3), 0.658(MIA Pa Ca-2), 1.949(CFPAC-1)and 1.077(Capan-1). We found that the expression of TRAIL-R1 was highest in human pancreatic carcinoma CFPAC-1 cell line, higher in MIAPa Ca-2 and panc-1 while low in BXPC-3 while do not express in Capan-1, also, TRAIL-R2 have the same trend, but no significant expression of TRAIL-R2 in BXPC-3 cells. Immunohistochemistry results indicated that the expression of TRAIL-R1 in pancreatic carcinoma 48% were lower than carcinoma tissues 75%(c2=7.43, P<0.05); Clinicopathological analysis demonstrated that TRAIL-R1 expression was associated with tumor differentiation(c2=8.60, P<0.05) and Tumor pathologic stage(c2=7.26, P<0.05) indicated lower expression of the TRAIL-R1 positive positively correlated with lower histological grade, advanced T status. In addition, our analysis found that TRAIL-R2, which in pancreatic cancer positive positively staining intensity was lower than the corresponding adjacent tissues(17%, 35%, c2 = 4.38, P <0.05), the poor degree of differentiation in pancreatic cancer, the lower expression of TRAIL-R2 in tumor tissue(c2 = 7.704, P <0.05).Conclusion DNT cells could secrete s TRAIL and it may be one of the mechanisms of DNT cells inhibit the proliferation of human pancreatic cancer cells by s TRAIL bind to TRAIL respects, which expressed by pancreatic cancer cell lines and tissues, to induced cell apoptosis.