CD40配体对结肠癌细胞株的体外抑制作用

研究免疫共刺激分子CD40在结肠癌细胞株的表达及重组可溶性人CD40配体(rshCD40L)对结肠癌细胞株的体外抑制作用。采用流式细胞仪分别检测4株结肠癌细胞(HCT116、Caco-2、SW48和COLO-320)CD40分子的表达,同时利用流式细胞仪检测γ干扰素(IFN-γ)对结肠癌细胞株CD40表达的影响。MTT法检测rshCD40L对结肠癌细胞的抑制作用,TUNEL法检测rshCD40L对结肠细胞的促凋亡作用,同时检测联合rshCD40L和IFN-γ抗结肠癌作用。结果流式细胞仪检测有3株结肠癌细胞(HCT116、Caco-2和SW48)表达CD40分子,IFN-γ能增强CD40+结肠癌细胞CD40分子的表达。rshCD40L能明显抑制CD40+结肠癌细胞的增殖作用(抑制率分别为HCT116:33.5%±5.5%;Caco-2:21.5%±3.6%;SW48:30.1%±4.2%),而对CD40-结肠癌细胞株(COLO-320)无明显抑制作用(P<0.05)。rshCD40L能诱导CD40+结肠癌细胞的凋亡作用(凋亡细胞百分比为HCT116:25.6%±4.52%;Caco-2:15.71%±2.27%;SW48:18.0%±3.7%),而对CD40-结肠癌细胞株(COLO-320)无诱导凋亡作用(P<0.05)。另外,IFN-γ能明显增强rshCD40L对结肠癌细胞的抑制增殖和促凋亡作用。重组人CD40配体与IFN-γ联合可显著诱导CD40阳性的结肠癌细胞凋亡,抑制其增殖,具有潜在的抗肿瘤作用。

The growth-inhibitory activity of CD40 ligand in colon cancer in vitro

To investigate the expression of CD40 molecule on colon cancer cell lines and to evaluate the growth-inhibitory effect of recombinant soluble human CD40L (rshCD40L) on colon cell lines,the expression of CD40 on colon cell lines (HCT116,Caco-2,SW48,and COLO-320) and the changes induced by interferon-γ(IFN-γ) were examined by flow cytometry.The growth-inhibitory activity of rshCD40L on colon cancer cell lines was examined by MTT assay and the proportion of apoptotic tumor cells was examined with TUNEL assay. Also,the growth-inhibitory effect of the combined activity of rshCD40L and IFN-γ was examined.As demonstrated by flow cytometry,the CD40 molecules could be detected on 3 strains of colon cancer cell lines (HCT116,Caeo-2 and SW48) and the molecular expression of CD40 on these cell lines could be enhanced by the action of IFN-γ.However,the proliferative activities of these cell lines were markedly inhibited by rshCD40L molecule with the growth-inhibitory activities of 33.5±5.5% for HCT116;21.5±3.6% for Caco-2 and 30.1±4.2% for SW48 cell lines respectively. Nevertheless,no apoptosis-induced effect could be detected in another CD40+colon cancer cell line COLO320 (P<0.05).In addition,IFN-γ could significantly enhance the activities of proliferation inhibition and induction of apoptosis. From these observations,it is evident that the combined activity of rshCD40L and IFN-γ inhibit proliferation and induce apoptosis of the CD40+colon cancer cell line,and thus has a potential anti-tumor effect.