GDNF增强Treg促进非小细胞肺癌细胞增殖和转移的作用研究

Glial cell line derived neurotrophic factor strengthen the ability of Treg to promote proliferation and metastasis of non-small cell lung cancer

为探讨胶质细胞源神经营养因子(glial cell line derived neurotrophic factor,GDNF)增强Treg促非小细胞肺癌(nonsmall cell lung cancer,NSCLC)细胞增殖和转移的作用,分离人外周血中淋巴细胞,用CD4、CD25和Foxp3标记外周血中的Treg;ELISA检测上清中IL-10和TGF-β的水平;EDU实验检测NSCLC细胞的增殖水平;Transwell侵袭实验检测NSCLC细胞的侵袭能力。流式结果显示GDNF能够显著促进淋巴细胞中Treg比例的上升,相对于对照组,低浓度GDNF组Treg的比例增加了6.37倍,相对于低浓度组,高浓度组Treg的比例增加了1.17倍(P<0.05);ELISA结果显示GDNF促进Treg分泌IL-10和TGF-β,相对于对照组,低浓度组上清中IL-10和TGF-β的浓度分别增加了4.92倍和4.86倍,相对于低浓度组,高浓度组IL-10和TGF-β的浓度分别增加了108%和119%(P<0.05);EDU实验结果显示GDNF能增强Treg促NSCLC细胞增殖作用,相对于Treg组,低浓度和高浓度组NSCLC细胞的增殖率分别上升了77%和144%(P<0.05);Transwell实验结果显示GDNF能增强Treg促进NSCLC细胞侵袭的作用,相对于Treg组,低浓度和高浓度GDNF组侵袭细胞的比例分别上升了2.86倍和5.83倍(P<0.05);IL-10和TGF-β的阻断均能部分降低GDNF增强Treg促进NSCLC细胞增殖和侵袭的作用(P均<0.05)。总之,GDNF能够通过诱导Treg的增殖,进而促进NSCLC细胞增殖和侵袭。

The purpose of this study was to investigate the role of glial cell line derived neurotrophic factor（GDNF）in enhancing the ability of Treg cells to promote proliferation and metastasis of non-small cell lung cancer.Human peripheral blood lymphocytes cells were separated,and regulatory T cells（Treg cells）were labeled with CD4,CD25 and Foxp3;ELISA was used to detect the levels of IL-10 and TGF-βin the supernatant;EDU tests were used to detect the proliferation of cells;Transwell invasion test was used to detect the metastatic ability of NSCLC cells.The results of flow cytometry showed that GDNF could significantly increase the proportion of Treg in lymphocytes,compared with the control group,the proportion of Treg cells in low concentration GDNF group increased by 6.37 times,in high concentration GDNF further increased the proportion of Treg cells by 1.17 time（P〈0.05）compared to the low concentration GDNF group.ELISA results showed that GDNF promoted Treg cells secreting IL-10 and TGF-β,compared with the control group,the concentration of IL-10 and TGF-βin the supernatant of low concentration GDNF group were increased by 4.92 and 4.86 times respectively.Compared with the low concentration GDNF group,TGF-βand IL-10 concentrations in high concentration GDNF group were increased by 108% and 119%（P〈0.05）respectively;EDU experimental results showed that the GDNF could enhance Treg cells＇ ability to promote nonsmall cell lung cancer cell proliferation,compared with the control group,NSCLC cell proliferation rates in low and high concentration GDNF group were increased by 77% and 144%（P〈0.05）respectively;Transwell results showed that the GDNF could enhance NSCLC cell invasion,compared with control group,the proportion of invasive cells in low concentration and high concentration GDNF group were increased by 2.86 and 5.83 times respectively（P〈0.05）.The blocking of IL-10 and TGF-βcould partly reduce the effect of GDNF enhanced-Treg on the proliferation and invasion of NSCLC cells（P〈0.05）.In conclusion,GDNF could induce Treg cell proliferation,further promote proliferation and invasion of NSCLC cells.