| 摘要: |
| 目的研究转化生长因子-茁1（TGF-茁1）促进胃癌侵袭能力，诱导胃癌MKN-45细胞发生上皮间质转化（EMT）的机制及其与PI3K/Akt信号通路调控肿瘤起始细胞标志物CD133表达的关系。方法设空白对照，应用TGF-茁1处理MKN-45细胞(TGF-茁1处理组)，观察其对细胞形态学的影响；Transwell检测细胞侵袭能力的改变；RT-PCR及Westernblot检测细胞EMT相关因子Snail、E-cadherin、N-cadherin、p-Akt及CD133的表达水平；PI3K特异性抑制剂LY294002预处理后再应用TGF-茁1处理细胞（TGF-茁1+LY294002处理组），检测p-Akt与CD133表达水平的变化；免疫磁珠分选CD133+与CD133-亚群细胞，检测CD133+组与CD133-组细胞EMT相关因子的表达差异。结果TGF-茁1处理72h后，TGF-茁1处理组细胞由上皮形态转化为间质形态。TGF-茁1处理组Snail、N-cadherinmRNA、蛋白表达水平均高于对照组（均P＜0.05）；而E-cadherinmRNA、蛋白表达水平均低于对照组（均P＜0.05）；Transwell检测发现TGF-茁1处理组穿膜细胞数高于对照组（P＜0.05）；TGF-茁1处理组p-Akt蛋白与CD133mRNA、蛋白表达水平均高于对照组（均P＜0.05）；TGF-茁1+LY294002处理组p-Akt蛋白与CD133mRNA、蛋白表达水平均低于TGF-茁1处理组（均P＜0.05）。CD133+组Snail、N-cadherinmRNA、蛋白表达水平均高于CD133-组（均P＜0.05），而E-cadherinmRNA、蛋白表达水平均低于CD133-组（均P＜0.05）。结论TGF-茁1诱导胃癌细胞发生EMT，并通过PI3K/Akt信号通路调控CD133表达从而增强胃癌MKN-45细胞的侵袭能力。 |
| 关键词: TGF-茁1 胃 新生物 / 癌 MKN-45 上皮间质转化 CD133 PI3K/Akt |
| DOI： |
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| TGF-茁1 induces epithelial-mesenchymal transition in gastric cancer cells and its mechanism |
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ZHONG Zhifeng, DU Jinlin, WANG Jianping, JIN Xihan, DAI Jian
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Jinhua Municipal Central Hospital
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| Abstract: |
| Objective To investigate the effect of TGF-茁 1 on the epithelial-mesenchymal transition (EMT) in gastric
cancer cells and its mechanism. Methods Human gastric cancer MKN-45 cells were treated with TGF-茁 1. The micromorphological changes were observed and the invasive potency of MKN-45 cells were examined with Transwell method; the mRNA and protein expressions of Snail, E-cadherin, N-cadherin, p-Akt and CD133 were detected by RT-PCR and Western blot, respectively. The MKN-45 cells were pretreated with PI3K special inhibitor LY294002, then treated with TGF-茁1, and the expressions of p-Akt and CD133 were detected. CD133+ and CD133- cells were sorted by MACS and the expression of EMT-related proteins were measured and compared between two subsets of MKN-45 cells. Results The micromorphology of culture cells was changed to mesenchymal profiles 72h after TGF-茁1 treatment; and the mRNA and protein expression levels of Snail and N-cadherin were higher than those in control group (P ＜0.05), the mRNA and protein expression levels of E-cadherin were significantly lower than those in control group (P＜0.05). The invasion ability of TGF-茁 1 treated group was higher than that of control group (P＜0.05). The relative protein expression levels of p-Akt and CD133 in TGF-茁1 treated group were higher than those in control group(P＜0.05). In LY294002 pretreated cells, the p-Akt and CD133were down-regulated(P＜ 0.05). The relative mRNA and protein expression levels of Snail, N-cadherin in CD133+ group were higher than those in CD133- group (P＜0.05), the relative mRNA and protein expression levels of E-cadherin in CD133+ group were lower than those in CD133- group(P＜0.05). Conclusion TGF-茁1 can induce EMT in MKN-45 cells, enhance the invasion ability of MKN-45 cells and up-regulate the expression of CD133 via PI3K/Akt pathway. |
| Key words: TGF-茁1 Stomach Neoplasma/cancer MKN-45 EMT CD133 PI3K/Akt |
