DNT细胞的扩增及其对胰腺癌Panc-1细胞的影响

Effects of ex vivo expanded DNT cells on growth of Panc-1 cells

目的探讨TCR+CD3+CD4-CD8-(DNT)双阴T细胞对人胰腺癌细胞系Panc-1的影响。方法应用抗体吸附法扩增人外周血DNT细胞,采用流式细胞仪检测所得细胞表型,选择多组对照观察DNT细胞对裸鼠荷瘤的生长曲线和抑瘤率的影响,应用RT-PCR检测不同浓度DNT细胞与Panc-1细胞体外培养后Fas/FasL的表达情况。结果经过12 d左右的培养,每孔细胞数量可达1.0×106个以上。流式细胞仪鉴定细胞表型为CD3+CD4-CD8-。与阴性对照组比较,实验组和阳性对照组对裸鼠荷瘤的肿瘤生长曲线及抑瘤率的差异有统计学意义,实验组的抑瘤率达44.75%。DNT细胞与胰腺癌细胞共培养后,胰腺癌细胞在mRNA水平的Fas表达量明显上升,差异有统计学意义;FasL表达略有下调,差异无统计学意义。结论 DNT细胞能抑制胰腺癌Panc-1细胞株增殖,Fas/FasL途径为其作用机制之一。

Objective To find out the effects of CD3^＋ CD^4- CD8^- double negative T cells on suppression of pancreatic cancer in vitro and vivo. Methods DNT cells were expanded in vivo based on a protocol of antibody absorption, and the cell surface molecules were identified by flow cytometry. The growth curve and inhibitory rate of Panc-1 xenografted nude mice was observed. The expression of Fas/FasL of Panc-1 with different concentrations of DNT cells was detected by RT-PCR. Results A mean of 1.0 × 10^6/hole was obtained after 12 day＇ culture of the expanded cells. In animal experiments, both DNT cells and celecoxib could suppress the growth curve and inhibitory rate of Panc-1 xenografted nude mice, and differences had statistical significance. After 48 hours co-culture with DNT cells, the Fas up-regulated their expressions, with statistically significant differences, while the FasL down-regulated their expressions. Conclusion DNT ceils could inhibit the growth of Panc-1, with Fas/FasL pathway as one of the important mechanisms.