| Objective To investigate the expression of CXCL16 in pancreatic cancer tissue and pancreatic cancer cell lines,and study the factors which effect on CXCL16 and the effects of CXCL16 on the biological behaviors of pancreatic cancer cell lines.Methods The expression of CXCL16 was detected by immunohistochemistry in 30 cases of pancreatic cancer tissue and paracancerous tissue,to analyse the clinical pathology relationship such as TNM'stage,metastastic lymph nodes and Broder' grades.The expression of transcription and protein of CXCL16 were detected by RT-PCR and ELISA.Exponentially growing PANC-1ASPC-1 cells exposed to different concentration of TNF-a and IL-1βindividually(10,50,100,200ng/ml) were determined the OD value of CXCL16,and cell lines without TNF-a and IL-1βtreament was used as control.Cell lines which exposed to the 200ng/ml concentration of TNF-a and IL-1βtogether were determined OD value of CXCL16,and cell lines with 200ng/ml concentration was used as the control.Exponentially growing PANC-1 and ASPC-1 cells were exposed to different concentration of rhCXCL16 (50,100,200ng/ml) and CXCL16 antibody,and cell lines without rhCXCL16 treament were used as the control.The proliferation was determined by MTT method,adhesion rate was determined by Matrigel matrix,invasion and migration of cell lines were assayed by Transwell chamber. Results Positive expression of CXCL16 was detected in 80.00% pancreatic cancer tissue and in 26.77% paracancerous tissue (P<0.005).The expression of CXCL16 was correlated with TNM'stage, metastastic lymph nodes (P<0.05) and has no association with Broder'grades.The CXCL16mRNA gelatin strap was founded by RT-PCR.On the 106/ml concentration of cell,the OD value of CXCL16 of PANC-1 and ASPC-1 cell lines was 215.3±14.9 and 170.0±11.6 individually.On the concentration of 200ng/ml of TNF-aand IL-1B,the OD value of CXCL16 of PANC-1 was 460.3±46.4 and 421.5±44.6,and which of ASPC-1 was 421.5±44.6 and 367.7±46.0 respectively,which significantly higher than 90.7±3.5 and 83.2±4.5 in the control group (P<0.05).Both the 200ng/ml concentration of TNF-aand IL-1β,the OD value of CXCL16 was 481.2±50.0 and 433.3±45.0 respectively,and without significant difference than the control.After 100ng/ml rh CXCL16 treament,proliferation,adhesion rate,invasion and migration of PANC-1 were 0.227±0.021,(91.4±8.6)%,1.246±0.216,1.361±0.276 respectively,and the adhesion rate,invasion and migration were significantly higher than (20.6±3.2)%,0.259±0.013,0.199±0.008 in the control group (P<0.05),and without significant effect on proliferation.After 200ng/ml rhCXCL16 treament, adhesion rate,invasion and migration of PANC-1 cells were further improved.After 100ng/ml rhCXCL16 treament,proliferation,adhesion rate,invasion and migration of ASPC-1 were(92.1±6.3)%,1.511±0.174,1.600±0.208 respectively,and the adhesion rate,invasion and migration were significantly higher than (21.6±3.8)% 0.245±0.012,0.190±0.010 in the control group (P<0.05),and without significant effect on proliferation.After 200ng/ml rhCXCL16 treament, adhesion rate,invasion and migration of ASPC -1 cells were further improved.Conclusions The level of CXCL16 in tissue of pancreatic cancer was obviously higher than those of paracancerous tissue group,and it involve development of pancreatic cancer.Through the transcription and protein level,CXCL16mRNA and protein were expressed in PANC-1 and ASPC-1 respectively.TNF-aand IL-1βcould enhance the secretion of CXCL16 respectively.However,both factors could not significantly enhance the expression of CXCL16 than the single factor.rhCXCL16 could enhance the ability of adhesion,invasion and migration of PANC-1 and ASPC-1 cell lines,but it could not significantly enhance the ability of proliferation. |
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