Gastric cancer stem cells in tumor invasion and metastasis and its influence on angiogenesis ability

doi:10.3969/j.issn.2095-4344.2016.32.004

Abstract

Abstract:

BACKGROUND: Gastric cancer stem cells involved in the neoadjuvant chemotherapy and conventional treatment are closely associated with relapse of gastric cancer. However, this conclusion has not yet been confirmed.
OBJECTIVE: To investigate the effects of gastric cancer stem cells in tumor invasion and metastasis and its effect on angiogenesis ability.
METHODS: We prepared nude mouse models of gastric cancer to isolate and culture gastric cancer stem cells. Harvested gastric cancer stem cells were detected in cell scratch test, ring test, inhibition rate test, cell migration test and tumorigenicity test.
RESULTS AND CONCLUSION: After 7 days of culture, the cells exhibited adherent growth but a lack of regularity that most cells were in a tadpole shape. In the cell scratch test, the scratch width was significantly different at 0 and 24 hours (P < 0.05). Under an inverted microscope, the cells were found to form a ring in the ring test. The 50% inhibiting concentration of gastric cancer stem cells induced by oxaliplatin was significantly lower than that induced by 5-fluorouracil (P < 0.05). The number of cells passing through the basilar membrane was significantly increased after cell migration (P < 0.05). After implantation of gastric cancer stem cells, the gastric tissue quality was significantly higher than that in normal nude mice of gastric cancer (P < 0.05). These findings indicate that gastric cancer stem cells responsible for tumor invasion and migration have stronger angiogenesis ability.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Tissue Engineering, Stomach Neoplasms

CLC Number:

R394.2

Zhang Li-xian, Zhang Ning, Yuan Shuang-zhen, Chen Yu-mei, Su Xin-ai, Sun Jian-shun, Wang Li-hua, Lin Mei. Gastric cancer stem cells in tumor invasion and metastasis and its influence on angiogenesis ability[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(32): 4738-4744.

Figures/Tables 1

2.1 实验动物数量分析裸鼠30只全部进入结果分析数量，无脱落。 2.2 胃癌干细胞形态观察胃癌干细胞分离后进行培养，细胞悬浮生长，培养7 d后贴壁生长，悬浮生长细胞相对较少，多为圆形、椭圆形；贴壁生长细胞相对较多，且缺乏规律性，细胞多为蝌蚪形，见图2。 2.3 细胞划痕实验结果胃癌干细胞在划痕后0，24 h采用显微镜观察划痕的宽度，结果显示：植入胃癌干细胞前划痕宽度为(0.750±0.023) mm，24 h后划痕宽度为(0.310±0.016) mm，不同时间段划痕宽度差异有显著性意义(P < 0.05)，倒置显微镜下可见胃癌干细胞位置发生率了明显的变化，见图3。 2.4 胃癌干细胞成环实验结果调整胃癌干细胞浓度为1×104个接种在96孔板中，培养24 h后观察成环数量，结果显示：胃癌干细胞成环数量为(7.20±1.47)个，通过倒置显微镜观察成环形态，胃癌干细胞注射后能形成完整的环，见图4。 2.5 细胞抑制率实验结果采用MTS方法检测奥沙利铂以及5-氟尿嘧啶对胃癌干细胞细胞活性的影响。结果显示：奥沙利铂为胃癌干细胞的半抑制浓度为(0.67±0.17) mg/L，而5-氟尿嘧啶的胃癌干细胞半抑制浓度为(2.23±0.56) mg/L，两者相比较差异有显著性意义(P < 0.05)。 2.6 胃癌肿瘤干细胞转移性实验结果结晶紫染色结果显示，胃癌肿瘤干细胞穿过基底膜细胞数为(172.60±12.28)个，细胞转移前为(85.20±6.57)个，两者比较差异有显著性意义(P < 0.05)，见图5。 2.7 胃癌肿瘤干细胞成瘤实验结果将1×106个细胞接种在裸鼠背部，在同等条件下喂养4周后麻醉处死裸鼠，测量胃癌组织质量发现，胃癌干细胞植入后胃癌组织质量为(0.624±0.036) g，普通胃癌裸鼠为(0.165± 0.011) g，两者差异有显著性意义(P < 0.05)，胃癌转移瘤组织变化见图6。"

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