CD40与其配体的连接对于肺癌细胞调节功能的研究

目的 研究CI40与其配体(CD40L)的连接对于肺癌细胞的调节功能,评价CI40作为治疗靶抗原的潜能。方法 通过基因克隆技术、Western蛋白印迹、流式细胞术等研究CD40L对于7株肺癌细胞系(包括1例转染CD40 cDNA细胞系)的细胞表型、细胞生长、细胞周期和凋亡的影响。结果 CD40L使高表达CI40的肺癌细胞系上主要组织相容性抗原—Ⅰ类分子(MHC—Ⅰ)、CD54和Fas分子的表达增强,上皮生长因子受体(EGFR)的表达减弱;CD40L作用5d后,CD54平均荧光强度(mean值)大约上调了4—10倍,MHC—Ⅰ上调了4—8倍,Fas上调了2．7—6倍,EGFR下调了2—3倍。细胞生长受到抑制,第5天受抑最明显(受抑率≥30％);CD40L停用后,这些变化有所恢复;CD40L对于CD40中低表达和不表达的细胞系无明显影响;所有细胞系均无凋亡发生。结论 CI40高表达肺癌细胞上CI40的表达具有免疫导向治疗靶抗原的潜能。

Functional significance of CD40 and CD40 ligand linking on human lung cells

OBJECTIVE: To determine the possible functional significance of CD40 and CD40 ligand (CD154) linking on human lung carcinomas and to assess the potential of CD40 as a therapeutic target. METHODS: We evaluated the effect of CD40L on the surface expression of major histocompatibility complex class I (MHC-I), Fas, bcl2, CD54, epidermal growth factor receptor (EGFR), p-glucoprotein (PGP), lung related protein (LRP), cell cycle, cell apoptosis and growth kinetics of 7 lung cancer cell lines (including 1 CD40-transfected cell line GLC-82/CD40) by gene cloning, Western blotting, and flow cytometry etc. RESULTS: Significant increased expression of MHC-I, CD54 and Fas was observed in 4 tumor lines expressing high levels of CD40 after CD40L (0.1 micro g/ml) linked to CD40. CD40L (0.1 micro g/ml or greater) in the fifth day was found to significantly inhibit the proliferation of 4 cell lines expressing high levels of CD40, decreased the percentage of aneuploid cell, and inhibited S-phase cells entering G2/M phase. The effect of CD40 cross-linking was reversible. CD40-moderate and low and negative tumor did not respond to CD40L. All of 7 cell lines show no significant changes in apoptosis in either the experimental (CD40L pulsed) or control (medium only) cell cultures. CONCLUSION: Expression of CD40 on lung tumors cells expressing high level of CD40 (including CD40-transfected cells) may represent a potential therapeutic target.