同种肿瘤来源的HSP70-PC对原位的大鼠膀胱肿瘤的治疗研究

 目的 研究用来自亚高温盆腔区域热疗后原位大鼠膀胱肿瘤的热休克蛋白70-肽复合物（HSP70-PC）对原位大鼠膀胱肿瘤的治疗作用机制。方法 经尿道膀胱灌注MNU制备出大鼠膀胱肿瘤动物模型。将动物随机分为A（肿瘤对照）组、B（HSP70-PC治疗）组、C（肿瘤细胞裂解液治疗）组，每组5只。皮下注射治疗1个月后检测膀胱及肿瘤湿重，肿瘤分期分级，TUNEL法检测肿瘤细胞凋亡指数（AI），免疫组化检测肿瘤细胞增殖细胞核抗原指数（PCNA LI）、肿瘤组织中S-100蛋白阳性细胞率及CD8阳性细胞率、膀胱引流区域髂动脉及腹主动脉旁淋巴结中S-100蛋白阳性细胞率、脾脏S-100蛋白阳性细胞率及CD8阳性细胞率，ELISA法检测血清Rat IFN-γ含量。结果 膀胱及肿瘤湿重B组低于A、C组（P＜0.01）。A、C组间差异无显著性意义（P = 0.610）。B组肿瘤分期低于A、C组（P＜0.01）；A、C组间差异无显著性意义（P =1.00）。肿瘤分级各组间差异均无显著性意义（P =1.00）。B组肿瘤细胞AI、膀胱肿瘤组织中CD8阳性细胞率、血清Rat IFN-γ含量均高于A、C组（P＜0.01）；A、C组间差异均无显著性意义（P =0.870，0.597，0.979）。脾脏S-100蛋白阳性细胞率及CD8阳性细胞率由高到低依次为B、C、A组（P＜0.01）。肿瘤细胞PCNA LI、膀胱肿瘤组织中S-100蛋白阳性细胞率、淋巴结中S-100蛋白阳性细胞率三组之间差异均无显著性意义（P =0.808，0.718，0.847）。结论 皮下注射HSP70-PC可以上调原位的膀胱肿瘤大鼠的细胞免疫状态，促进肿瘤凋亡，但对肿瘤细胞增生无影响。

The pathologic and immunologic changes of orthotopic rat bladder tumor were induced by HSP70-peptides complexes derived from autologous tumor

Objective To explore the treatment mechanism of HSP70-peptides complexes (HSP70-PC) derived from autologous tu-mor after heated by non-invasive pelvic cavity regional moderate hyperthermia, we applied the protein to rats containing orthotopic bladder tu-mor. Methods 1. Establishment of orthotopic rat bladder tumor model: Six-week-old female inbred line F344 rats of 20 totals were instilled methyl-nitroso-urea transurethrally once every other week for 6 times. Then 5 rats were continuous bred 4 weeks, and the other 15 rats were continuous bred 8 weeks respectively. 2. HSP70-PC has been purified from rat bladder tumor heated by non-invasive pelvic cavity regional moderate hyperthermia by our improved methods in our previous study. 3. Therapy to orthotopic rat bladder tumor: The 15 rats were separated randomly to group A (control), group B (HSP70-PC therapy) and group C (tumor lysate therapy). 4. The wet weight of bladder containing tu-mors was weighed after 4 weeks. The pathologic grade and stage of tumor were detected. The positive cells of S-100 Protein and CD8 immerged in tumor, the positive cells of S-100 Protein and CD8 in lymphonode of draining domain around bladder, the positive cells of S-100 Protein and the positive cells of CD8 in spleen and PCNA of tumor cells were detected by immunohistochemistry. Apoptosis of tumor cell were detected by TUNEL. The quantity of Rat IFN-γ was detected by ELISA. Results 1. The weight of bladder containing tumors of group B were significant lower than that of group A and C (P <0.01). But there was non-significant difference between group A and C (P =0.610). The stages of tumor of group B were significant lower than that of group A and C (P <0.01). But there was non-significant difference between group A and C (P =1.00). There were non-significant differences of grade of tumor among groups A, B and C (P =1.00). The ratio of apoptosis index of tumor cells, CD8 positive cells in tumor and Rat IFN-γ quantities of group B were significant higher than that of group A and C (P <0.01). But there were non-significant difference between group A and C (P =0.870, 0.597, 0.979). The descending ranks of the ratio of S-100 protein positive cells and the ratio of CD8 positive cells in spleen were group B, C and A (P <0.01). There were non-significant differences among three groups of PCNA labeled indexes of tumor cells, the ratio of S-100 Protein positive cells immerged in tumor and positive cells of S-100 Protein in lym-phonode of draining domain around bladder (P =0.808, 0.718, 0.847). Conclusions HSP70-PC can improve both cellular immunity of rat containing orthotopic bladder tumor and tumor cell apoptosis. Nevertheless it could not affect proliferation of tumor cells.