CAJ | 학술논문

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.
목적 관찰이이점단백(MTDH)/성형세포상조기인-1(AEG-1)위파점적기인역묘유도면역억제소서전렬선암생장、전이적작용.방법 장C57BL/6웅성소서(6～8주령)수궤분위3조(n=20),분별급여린산염완충액(PBS)、Pub、Pub-MTDH/AEG-1적감독사문씨균(1×108cfu)관위사복면역；매주면역1차,공3차.최후1차면역후1주,재3조소서전렬선원위(좌、우배측협)접충105 RM-1세포구건전렬선원위이식류모형,유도자발분강、복막후림파결전이；기록각조소서종류대소급분강、복막후림파결전이수[이소목소-이홍(HE)염색위준],통과면역조직화학방법검측원위종류중CD8、혈관내피생장인자(VEGF)、림파관내피투명질산수체-1(LYVE-1)적표체、용원위말단전이매표기(TUNEL)법검측종류세포조망.결과 pUb-MTDH/AEG-1DNA역묘조여Pub、PBS조비교능현저격발CD8+T세포급세포독성T림파세포(CTL)반응；유효억제소서원위전렬선암생장,MTDH조종류체적(0.248 ±0.102)명현소우Pub조(1.475 ±0.314) (P ＜0.01)；량조소서분강、복막후림파결전이솔분별위77.78％、35.7％(P＜0.05)；병현저증강원위종류중CD8+분자적표체:MTDH조3.56±0.85、Pub조5.12±0.90 (P＜ 0.05),촉진종류세포조망:MTDH조(39.60±3.28)％、Pub조(16.18±2.52)％(P＜0.01).결론 MTDH/AEG-1위파점적기인역묘재소서예방면역모형중,능유효유도궤체특이성면역응답,증강세포면역급체액면역,억제전렬선암증식、침습,촉진종류세포조망,연장하류소서생존시간.
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.