摘要
目的探讨肿瘤坏死因子相关凋亡诱导配体(sTRAIL)与其受体TRAIL-R1在DNT细胞杀伤胰腺癌细胞中的作用及意义。方法采用抗体吸附法培养外周血中分离的DNT细胞,ELISA法检测其细胞培养上清液中的sTRAIL水平。采用qPCR、Western blot法检测TRAIL-R1在5株胰腺癌细胞株中的表达,免疫组织化学法检测48例胰腺癌组织标本中TRAIL-R1的表达情况,分析其与病理参数之间的关系。结果 DNT细胞数目逐渐增多,其培养液中sTRAIL与对照组相比浓度明显升高(t=17.24,P<0.05)。qPCR检测受体TRAIL-R1在5株胰腺癌细胞中均有表达,Western blot法检测TRAIL-R1受体表达发现在胰腺癌细胞系CFPAC-1中表达高,在MIA PaCa-2和panc-1中表达较高,BXPC-3中表达较低,在Capan-1细胞系中不表达。TRAIL-R1在胰腺癌组的阳性表达率及染色强度低于相应的癌旁组织(χ2=7.43、12.48,P<0.05);TRAIL-R1的表达与染色强度有关(χ2=12.48,P<0.05)。结论外周血来源的DNT细胞可分泌sTRAIL,sTRAIL与胰腺癌细胞和组织所表达的受体TRAIL-R1结合诱导细胞凋亡,可能是DNT细胞抑制胰腺癌细胞增殖的机制之一。
Objective To investigate the role and significance of the soluble tumor necrosis factor-related apoptosis- inducing ligand （sTRAIL） binding to its receptor TRAIL-R1 in DNT cell suppressing, Methods. DNT cells could be cultured according a novel of effective protocol of antibody adsorption, ELISA methods were used to measure the levels of sTRAIL in the culture supernatants. Real time RT-PCR, Western blot were employed to compare the ex- pression of TRAIL-R1 in 5 pancreatic cancer cell lines and the expression of TRAIL-R1 in 48 cases of pancreatic ductal adenocarcinoma tissues could be observed by immunohistochemistry, we evaluated the association of TRAIL- R1 with clinical pathological parameters. Results The number of cells gradually was improved and serum sTRAIL were higher in supernatants group than that in control group （t = 17.24 ,P 〈 0. 05 ）. High level of TRAIL-R1 was detected in 5 pancreatic cancer cell lines from transcript. We found that the expression of TRAIL-R1 was highest in human pancreatic carcinoma CFPAC-1 cell line, higher in MIA PaCa-2 and pane-1 while low in BXPC-3 while do not express in Capan-1. Immunohistochemistry results indicated that the expression and stainingintensity of TRAIL- R1 were lower than carcinoma tissues （ χ2 = 7.43, P 〈 0. 05 ） and expression positively correlated with histological grade and T status （ χ2 = 12.48,P 〈 0. 05 ）. Conclusion DNT ceils could secrete sTRAIL and it may be one of the mechanisms of DNT cells inhibiting the proliferation of human pancreatic cancer cells by sTRAIL bind to TRAIL-R1, which is expressed by pancreatic cancer cell lines and tissues, to induced cell apoptosis.
出处
《安徽医科大学学报》
CAS
北大核心
2014年第11期1613-1617,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81071985)
安徽省国际科技合作项目(编号:10080703038)
