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目的:探讨胃癌乳斑微转移模型在胃癌干细胞及前驱细胞研究中的意义。方法:胃癌细胞系MFC,1×10~4细胞数量注射到小鼠腹腔内,72 h后取大网膜,收集乳斑微转移灶内的胃癌细胞团;以不同细胞数量进行NOD/SCID小鼠皮下注射,观察细胞的致瘤能力,移植瘤HE染色,免疫组化分析CEA、CK20表达;免疫细胞化学分析乳斑微转移灶内的胃癌细胞团干细胞相关标志物的表达。结果:皮下注射16周后,微转移灶内的胃癌细胞团组中,皮下注射数量为5×10~2个细胞有1个部位产生肿瘤组织:注射数量为1×10~3个细胞时,5个部位均能产生肿瘤组织。对照组皮下注射为5×10~4个细胞时,仅1个部位出现肿瘤,而注射部位全部出现肿瘤组织,需要5×10~6个以上的细胞数量。两组皮下肿瘤组织分别经HE染色和免疫组化分析CEA、CK20表达,与对照组比较无统计学差异。微转移灶内细胞团高表达CD133和CD44,而CD324的表达则明显下降。结论:胃癌乳斑微转移模型内存在丰富的胃癌于细胞及前驱细胞;大网膜乳斑微环境具有富集胃癌干细胞及前驱细胞的作用。CD133~+、CD44~+和CD324~-可能是胃癌干细胞的表面标志物。
Objective: To investigate the significance of gastric mucosal micrometastasis in gastric cancer stem cells and precursor cells. Methods: The gastric cancer cell line MFC was injected into the peritoneal cavity of mice at a dose of 1 × 10 ~ 4 cells. After 72 hours, the omentum was taken and the gastric cancer cell mass in the milk spot micrometastasis was collected. The NOD / SCID The mice were injected subcutaneously to observe the tumorigenicity of the cells. HE staining was performed on the tumor cells. The expressions of CEA and CK20 were analyzed by immunohistochemistry. The expressions of stem cell markers in gastric cancer cells were analyzed by immunocytochemistry. Results: After subcutaneous injection for 16 weeks, the number of subcutaneous injection of 5 × 10 ~ 2 cells in one group of tumor cells in the group of gastric cancer cells in the micrometastasis: 1 × 10 ~ 3 cells, 5 parts can produce tumor tissue. Control group subcutaneous injection of 5 × 10 ~ 4 cells, only one tumor site, and the injection site of all tumor tissue, the need for more than 5 × 10 6 cells. The subcutaneous tumor tissues of the two groups were respectively analyzed by HE staining and immunohistochemistry for the expression of CEA and CK20, which showed no significant difference compared with the control group. Micrometastasis in the cell mass expression of CD133 and CD44, while the expression of CD324 decreased significantly. Conclusion: There are abundant gastric cancer cells and progenitor cells in gastric micrometastasis model; omental milky microenvironment has the function of enriching gastric cancer stem cells and precursor cells. CD133 ~ +, CD44 ~ + and CD324 ~ - may be the surface markers of gastric cancer stem cells.