rhCD40L联合化疗药物对乳腺癌细胞株生物学行为的影响  

Study on the biological effects of CD40 ligation on human breast cancer cell lines

作　　者：施勤[1] 陈永井[1] 谢芳[1] 马泓冰[1] 季玉红[1] 於葛华[1] 王勤[1] 葛彦[1] 张学光[1] 

机构地区：[1]苏州大学医学生物技术研究所,苏州215007

出　　处：《现代免疫学》2005年第1期5-10,共6页Current Immunology

基　　金：国家"973"基金资助项目(2001CB51003)

摘　　要：为了研究人重组CD40L(rhCD40L)对乳腺癌细胞株MDA-MB-231、MDA-MB-435细胞生物学行为的影响,运用多种细胞生物学、分子生物学方法检测细胞生长、细胞周期、细胞膜分子、凋亡相关基因Bcl-Xl、Bax及趋化因子RANTESmRNA水平的改变,并观察了rhCD40L联合IFN-γ、阿霉素(ADM)对MDA-MB-231细胞增殖的影响。结果发现rhCD40L抑制MDA-MB-435、MDA-MB-231细胞增殖,与剂量呈负相关(P<0.05);两株乳腺癌细胞分别在CD40配基化(ligation)作用48、72h进入G1期细胞阻滞;MDA-MB-231细胞CD54、CD95(Fas)、CD120b等分子表达均显著上调,CD95L(FasL)表达下调(P<0.05);MDA-MB-435细胞CD49e、CD95L、CD120a、HLA-DR等分子有显著性变化。两株乳腺癌细胞经rhCD40L作用4、24h后,RT-PCR半定量Bax/Bcl-Xl的比率、RANTES的表达与对照组相比均上调。rhCD40L联合IFN-γ、阿霉素对MDA-MB-231细胞增殖有较强的抑制作用(P<0.05)。由此表明,CD40信号影响乳腺癌细胞周期及TNF家族成员及MHCII类分子的表达,上调凋亡相关基因Bax/Bcl-Xl的比值和趋化因子RANTESmRNA水平,直接或间接介导抗肿瘤作用。同时,rhCD40L还可增强肿瘤细胞对细胞因子和化学药物的敏感性。To investigate the influence of recombinant human CD40 ligand(rhCD40L) on the biological behaviors of human breast cancer cell lines MDA,MB,231 and MDA,MB,435,the changes of cell proliferation,cell cycles,cell membrane molecules,apoptosis,related genes Bcl,Xl and Bax and level of chemokines RANTES mRNA were observed by means of many cell biology and molecular biology methods.Meanwhile,the effects of the combined use of rhCD40L with IFN,γ and ADM were assayed. The results showed that rhCD40L could inhibit the proliferation of the MDA,MB,435 and MDA,MB,231 cell lines,and the effects showed negative correlation with doses. In case of combined use with IFN,γ and ADM,the inhibitory activity was increased,and there was an increase in G1 phase of cell cycles with a significant decrease in the number of cells entering into S phase after the MDA,MB,231 and MDA,MB,435 cell lines were activated by rhCD40L for 48 and 72 hours respectively. An up,reffulatory effect of surface molecules,such as HLA,DR,CD95,CD120a was also observed. In addition,as demonstrated by RT,PCR,the semi,quantitative ratio of Bax/Bcl,Xl and the level of RANTES transcription up,regulated accordingly. These data suggest that the CD40 ligation with tumor cells can expand the anti,tumor immunity not only by way of direct cytotoxic effect to suppress the growth of tumor cells,but also by way of up,regulating adhesion molecules,co,stimulators and cytokines to rectifying the T cell immunity. Meanwhile,rhCD40 can also increase the sensitivity of tumor cells to the effects of cytokines and chemotherapeutics.

关 键 词：rhCD40L 乳腺癌细胞株 IFN-Γ 阿霉素 抗肿瘤 

分 类 号：R392.11[医药卫生—免疫学]