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目的 探讨CD40配基化(CD40 ligation)对肺癌细胞株NCI-H460、A549、SPC-A-1的增殖以及对细胞肿瘤坏死因子受体(TNFR)和肿瘤坏死因子受体相关因子(TRAF)表达谱的影响。方法 采用四氮唑盐(MTT)比色法检测CD40配体(CD40 ligand,CD40L)对肺癌细胞增殖的影响,免疫荧光标记和流式细胞术测定细胞表面CD40的表达以及CD40配基化后细胞TNFR和TRAF表达的改变。结果(1)肺癌细胞株NCI-H460、A549和SPC-A-1表面CD40表达分别为(89.2±3.2)％、(42.2±4.5)％、(2.3±0.3)％。(2)CD40配基化后能显著抑制NCI-H460、A549细胞的增殖(P<0.05),而不抑制SPC-A-1的增殖(P>0.05)。(3)CD40配基化可上调NCI-H460、A549细胞TNFRⅠ的表达,下调TNFRⅡ的表达(P<0.05),而不影响SPC-A-1细胞TNFR的表达。(4)CD40配基化可下调NCI-H460、A549细胞中TRAF2、3、5、6的表达(P<0.05),而不影响SPC-A-1中TRAFs的表达。结论 CD40配基化可抑制CD40表达阳性细胞株NCI-H460、A549细胞的增殖,并可引起细胞TNFR、TRAF表达谱的改变,提示CD40信号对CD40表达阳性的肺癌细胞生物学行为具有重要调节作用。
Objective To investigate the effects of CD40 ligation on the proliferation of lung cancer cell lines NCI-H460, A549 and SPC-A-1 as well as on the expression of tumor necrosis factor receptor (TNFR) and tumor necrosis factor receptor related factor (TRAF) Effects of spectrum. Methods The effect of CD40 ligand (CD40L) on the proliferation of lung cancer cells was detected by MTT assay. The expression of CD40 on the cell surface was measured by immunofluorescence staining and flow cytometry. TNFR and TRAF expression changes. Results (1) The expression of CD40 on lung cancer cell lines NCI-H460, A549 and SPC-A-1 were (89.2 ± 3.2)%, (42.2 ± 4.5)% and (2.3 ± 0.3)%, respectively. (2) CD40 markedly inhibited the proliferation of NCI-H460 and A549 cells (P <0.05), but did not inhibit the proliferation of SPC-A-1 cells (P> 0.05). (3) CD40 ligandization up-regulated the TNFRⅠexpression in NCI-H460 and A549 cells, down-regulated the expression of TNFRⅡ (P <0.05), but did not affect the expression of TNFR in SPC-A-1 cells. (4) CD40 ligandization down-regulated the expression of TRAF2,3,5,6 in NCI-H460 and A549 cells (P <0.05), but did not affect the expression of TRAFs in SPC-A-1. Conclusion CD40 ligand can inhibit the proliferation of CD40 positive cell lines NCI-H460 and A549 cells and induce the changes of the expression of TNFR and TRAF. These results suggest that CD40 signaling plays an important regulatory role in the biological behavior of CD40 positive lung cancer cells .