胃癌灶原位移植腹膜转移癌裸小鼠模型的建立及生物学性状鉴定

Establishment and identification of an orthotopic transplanted model of gastric carcinoma with peri- toneal carcinomatosis in nude mice

目的建立胃癌灶原位移植腹膜转移癌裸小鼠模型，鉴定其侵袭转移生物学特点。方法取11只BALB／C裸小鼠，将人MGC-803胃癌组织原位移植于胃窦部，建立裸小鼠人胃癌灶原位移植腹膜转移癌模型。测定裸鼠体质量、血常规，并采用肿瘤组织形态学及免疫组织化学方法观察肿瘤增生、凋亡及侵袭转移相关分子标志[B淋巴细胞／白血病（bcl）-2、bcl-6、原位末端转移酶标记（TUNEL）、细胞核增殖抗原（Ki-67）、CD34、D2-40、血管内皮生长因子（VEGF）、基质金属蛋白酶（MMP）-9]。结果模型制作成功率100％（11／11），生存期内裸鼠体质量平缓增加。常规病理学检测瘤块重量为210．5—379．0mg（中位数267．5mg），癌细胞异质性明显，微血管／淋巴管中癌栓易见，可见胃周淋巴结及肝脏转移。肿瘤增殖相关分子检测显示bcl-2阳性率为0．0％～10．4％（中位数为3．3％），bcl-6阳性率为0．0％-11．1％（中位数为3．3％），Ki-67阳性率为40．5％-48．6％（中位数为45．4％），TUNEL阳性率为0．2％-3．1％（中位数为0．9％），肿瘤侵袭转移相关分子检测肿瘤组织血管标记CD34观察到1—11（中位数为3）个脉管／视野，通过淋巴管标记D2-40观察到0—5（中位数为2）个脉管／视野，分泌VEGF的阳性细胞为68．5％～95．8％（中位数82．4％），MMP-9阳性细胞数为13．5％-50．0％（中位数为23．5％）。结论成功建立胃癌灶原位移植腹膜转移癌裸小鼠模型，并表现出了低分化胃腺癌的典型生物学特点。

Objective To establish an orthotopic transplanted model of gastric cancer with peritoneal carcinomatosis in nude mice and identify the pathological features of the model. Methods The homogenate of MGC-803 gastric cancer tissue was planted in gastric antrum of 11 BALB/c nude mice to establish the model of gastric cancer with peritoneal carcinomatosis. The general status was recorded. The routine morphological examination and immunohistochemistry were performed to detect tumor biological fea- tures related to proliferation, apoptosis, invasion and metastasis [ B lymphocytes/leukemia-2 （bcl-2）, bcl-6, Ki-67, TdT-mediated dUTP nick end labeling （TUNEL）, vascular endothelial growth factor （VEGF）, matrix metalloproteinase （ MMP）-9, CD34, and D2-401. Results The animal models were constructed successfully （11/11 ）. The body weight of nude mice was slowly increased during the model development. The orthotopic tumor weight was （210. 5-379.0） mg （median 267. 5 mg）. Cancer cell hetero- geneity, and microvascular/lymphatic tumor thrombus were obviously observed as well as perigastric lymph nodes and liver metastases. Tumor proliferation-related molecule detection showed that bcl-2 positive rate was 0. 0%-10. 4% （median 3.3% ）, bcl-6 positive rate was 0. 0%-11.1% （median 3. 3% ）, Ki-67 positive rate was 40. 5%-48.6% （median 45.4% ）, and TUNEL positive rate was 0. 2%-3. 1% （median 0. 9% ）. The number of tumor blood vessels marked with CD34 was 1-11 （median 3）/400 x field of vision. The number of lymphatics marked with D2-40 was 0-5 （median 2）/400 ~ field of vision. VEGF positive rate was 68.5%- 95. 8% （ median 82. 4% ）, and MMP-9 positive rate was 13.5% -50. 0% （ median 23. 5% ）. Conclusion The orthotopic transplanted model of gastric cancer with peritoneal carcinomatosis in nude mice was successfully established and reproduced the typical features of poorly differentiated adenocarcinoma.