目的探讨抗Cripto抗体(C13)对乳腺癌细胞的增殖抑制作用,以及抗Cripto抗体(C13)和羧胺三唑(CAI)合用是否有相加或协同作用,初步研究CAI与C13合用抑制肿瘤细胞增殖的作用机制。方法培养C13杂交瘤细胞,接种至SCID小鼠中,收集腹水,纯化后测定抗体效价。用不同浓度的C13抗体和CAI分别或共同处理MCF-7人乳腺癌细胞和66c14小鼠乳腺癌细胞。利用[^3H]-胸腺嘧啶掺入法分析CAI与C13对细胞DNA合成的影响。对细胞进行碘化丙啶染色,利用流式细胞仪评价两种药物对细胞凋亡的影响。结果在上述两种细胞系中,C13抗体无论是单用还是与CAI合用对肿瘤细胞的生长均有不同程度的抑制作用,且该作用呈剂量依赖性。流式细胞分析结果显示,C13单独或与CAI一同处理MCF-7细胞和66c14细胞后,以上细胞亚二倍体DNA的细胞比例升高,药物合用后该作用将进一步增加。结论针对Cripto17肽的抗体C13与CAI联用可以增强前者对乳腺癌细胞的抑制效应,这一作用可能与诱导细胞凋亡有关。
Objective To explore the inhibitory effect of anti-Cripto monoclonal antibody C13 on breast cancer cell, and itspotential additivity or synergy when combined with carboxyamidotriazole (CAI). The underlying mechanism of this combination was also discussed. Methods C13 hybridoma cell was cultured, and transferred into SCID mice to produce antibody. The ascites was collected, purified and determined for potency. Different concentrations of C13 and CAI were used to treat MCF -7 (human breast cancer cell line) or 66c14 (mouse breast cancer cell line), both concomitantly and independently. [ ^3H ] -TDR incorporation assay was employed to assess DNA synthesis. The extent of apoptosis was measured by flow cytometry after PI staining. Results C13 dose-dependently inhibited cell growth in all experiment settings for both MCF -7 and 66c14, no matter it was administered alone or in combination with CAI. Flow cytometry showed an elevated ratio of hypodiploid cell after C13 treatment, and this trend was more pronounced in C13 + CAI treatment group. Conclusion The introduction of CAI can enhance the effect of C13 ( monoclonal antibody against Cripto 17 - mer) on breast cancer cells in terms of cell growth inhibition. Its mechanism may involve apoptosis.