HSP70多肽复合物的抗肿瘤作用研究

目的： 研究HSP70多肽复合物对结肠癌荷瘤小鼠的免疫治疗作用，并探讨其抗肿瘤作用机制。方法：Colon26细胞经42 ℃水浴1 h、再分别于37 ℃培养8和12 h，并用反复冻融法筛选出含有高表达HSP70多肽复合物的瘤细胞裂解液。建立Colon26细胞的荷瘤小鼠模型，分别将含有高表达HSP70多肽复合物的瘤细胞裂解液 (实验组)，常规培养条件下制备的瘤细胞裂解液 (对照组Ⅰ)，以及PBS (对照组Ⅱ)分4次注射于小鼠皮下，观察小鼠体内成瘤性、生长情况及小鼠生存期。用流式细胞技术检测小鼠脾脏CD3+、CD4+和CD8+等T细胞亚型的变化。用ELISA法检测荷瘤小鼠血清中TGF-β1和IFN-γ水平的变化。结果：将高表达HSP70多肽复合物的瘤细胞裂解液注射于荷瘤小鼠后，小鼠的肿瘤生长速度、肿瘤体积和质量明显小于对照组Ⅰ与对照组Ⅱ，生存期也明显延长 (P<0.05)；实验组小鼠脾脏单个核细胞表面分子与对照组Ⅰ和对照组Ⅱ相比CD8+ T细胞显著增加 (P<0.05)，而CD3+和CD4+/CD8+ T细胞显著降低 (P<0.05)；实验组小鼠血浆中TGF-β1与对照组Ⅰ和对照组Ⅱ相比显著降低 (P<0.05)，但IFN-γ含量则显著升高 (P<0.05)。结论：含高表达HSP70多肽复合物的瘤细胞裂解液能抑制荷瘤小鼠瘤组织增长，延长荷瘤小鼠生存期，证明该多肽复合物通过调节免疫功能在小鼠体内发挥抗肿瘤作用。

The anti-tumor effects of HSP70 peptide complex

OBJECTIVE: To study on the immunotherapy effects of HSP70 polypeptide complex on colon cancer in mice，and to explore its antitumor mechanism. METHODS：The Colon26 cells were cultured at 42 ℃ for 1 h， then incubated at 37 ℃ for 8 and 12 h，and tumor cell lysates containing high expression of HSP70 polypeptide complex were prepared by repeated freezing and thawing methods. Tumor-bearing mice model was established with Colon26 cells. Tumor cell lysate containing high expression of HSP70 polypeptide complex (test group)，tumor cell lysate containing HSP70 polypeptide complex at 37 ℃ routine culture (control groupⅠ) and PBS (control groupⅡ) were repeatedly injected subcutaneously，in vivo tumorigenicity，growth and survival were monitored.　Expressions of CD3+， CD4+ and CD8+ T cells on the mice splenic monocuclear surface were analyzed by flow cytometry. ELISA was used to assess the production of TGF- β1 and IFN-γ in plasma of the tumor-bearing mice. RESULTS：After tumor-bearing mice were treated with tumor cell lysate containing high expression of HSP70 polypeptide complex，tumor growth，tumor volume and weight were significantly lower than control groupsⅠ and Ⅱ，the survival time was longer (P<0.05).Splenic mononuclear surface molecules of CD8+ T cells was significantly increased in the experimental group compared with the control groupsⅠand Ⅱ (P<0.05). CD3+ and CD4+/CD8+ T cells as well as TGF-β1 significantly reduced (P<0.05) in the plasma in mice of the experimental group compared with the control groupsⅠand Ⅱ，while IFN-γ level increased significantly (P<0.05). CONCLUSION：HSP70 polypeptide complex inhibited the tumor growth significantly，prolonged survival period，indicating that the peptide complex has obvious anti-tumor effects in mice by modulating immunology functions.