异黏蛋白/星形细胞上调基因-1靶点的基因疫苗诱导免疫抑制小鼠前列腺癌生长及转移

Metadherin/astrocyte elevated gene-1-based DNA vaccine suppresses progression and metastasis in prostate cancer of mice

目的 观察以异黏蛋白（MTDH）/星形细胞上调基因-1（AEG-1）为靶点的基因疫苗诱导免疫抑制小鼠前列腺癌生长、转移的作用.方法 将C57BL/6雄性小鼠（6～8周龄）随机分为3组（n=20）,分别给予磷酸盐缓冲液（PBS）、Pub、Pub-MTDH/AEG-1的减毒沙门氏菌（1×108cfu）灌胃饲服免疫；每周免疫1次,共3次.最后1次免疫后1周,在3组小鼠前列腺原位（左、右背侧叶）接种105 RM-1细胞构建前列腺原位移植瘤模型,诱导自发盆腔、腹膜后淋巴结转移；记录各组小鼠肿瘤大小及盆腔、腹膜后淋巴结转移数[以苏木素-伊红（HE）染色为准],通过免疫组织化学方法检测原位肿瘤中CD8、血管内皮生长因子（VEGF）、淋巴管内皮透明质酸受体-1（LYVE-1）的表达、用原位末端转移酶标记（TUNEL）法检测肿瘤细胞凋亡.结果 pUb-MTDH/AEG-1DNA疫苗组与Pub、PBS组比较能显著激发CD8＋T细胞及细胞毒性T淋巴细胞（CTL）反应；有效抑制小鼠原位前列腺癌生长,MTDH组肿瘤体积（0.248 ±0.102）明显小于Pub组（1.475 ±0.314） （P ＜0.01）；两组小鼠盆腔、腹膜后淋巴结转移率分别为77.78％ 、35.7％（P＜0.05）；并显著增强原位肿瘤中CD8＋分子的表达：MTDH组3.56±0.85、Pub组5.12±0.90 （P＜ 0.05）,促进肿瘤细胞凋亡：MTDH组（39.60±3.28）％、Pub组（16.18±2.52）％（P＜0.01）.结论 MTDH/AEG-1为靶点的基因疫苗在小鼠预防免疫模型中,能有效诱导机体特异性免疫应答,增强细胞免疫及体液免疫,抑制前列腺癌增殖、侵袭,促进肿瘤细胞凋亡,延长荷瘤小鼠生存时间.

Objective To study the metadherin （MTDH）/astrocyte elevated gene-1 （AEG-1） as targets for gene vaccine inducing immunosuppression mice prostate cancer growth and metastasis of action research.Methods Male C57BL/6 mice （6-8 weekes old） were randomly divided into three groups （n =20）,respectively,to phosphate buffered saline （PBS）,Pub,Pub-MTDH/AEG-1 attenuated salmonella （1 × 10s cfu） of lavage servo immune; Immune once a week,a total of immune three times.Last one week after the immunization,all in three groups of mice prostate in situ （right and left dorsal lobe） vaccinated 10s RM-1 cell to build orthotopic transplantation tumor model of prostate,induced spontaneous pelvic cavity and retroperitoneal lymph node metastasis; Record each group mice with tumor size,pelvic cavity and retroperitoneal lymph node metastasis number [hematoxylin and eosin （HE） staining shall prevail],CD8 in through immunohistochemical method to detect in situ tumor,the expression of vascular endothelial growth factor （VEGF） and lymphatic vessel endothelial hyaluronic acid receptor-1 （LYVE-1） case,tumor cell apoptosis with TdT-mediated dUTP nick end labeling （TUNEL） method.Results pUb-MTDH/AEG-1 DNA vaccine group compared to the pUb,the PBS group could significantly stimulate CD8 ＋ T cells and cytotoxic T lymphocyte （CTL） response; Effectively inhibit the growth of prostate cancer in situ in mice,MTDH group tumor volume （0.248 ± 0.102） compared with the Pub group （1.475 ± 0.314） （P ＜ 0.01）,pelvic cavity and retroperitoneal lymph node metastasis 77.78％,35.7％ （P ＜ 0.05） ; Significantly enhanced the expression of CD8 ＋ in the in situ tumor molecular：MTDH group （3.56 ± 0.85）,Pub group （5.12 ± 0.90） （P ＜ 0.05）,and promote tumor cell apoptosis：MTDH group （39.60 ± 3.28） ％,Pub group （16.18 ±2.52）％ （P＜0.01）.Conclusion MTDH/targets AEG-1 gene vaccine in mice prevention immune models,can effectively induce the body specificity immune response,enhance cellular immunity and humoral immunity,inhibit prostate cancer proliferation,invasion,and promote tumor cell apoptosis,extend a tumor-burdened survival time in mice.