| 舒 雄,刘辉琦,潘韵芝.ENO1在胃癌干细胞中表达及其与胃癌侵袭转移的相关性研究[J].中国肿瘤,2019,28(2):143-149. |
| ENO1在胃癌干细胞中表达及其与胃癌侵袭转移的相关性研究 |
| Expression of ENO1 in Gastric Cancer Stem Cells and Its Relation to Invasion and Metastasis of Gastric Cancer |
| 投稿时间：2018-04-12 |
| DOI：10.11735/j.issn.1004-0242.2019.02.A013 |
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| 中文关键词: 胃癌干细胞 ENO1 CD44 侵袭 信号通路 |
| 英文关键词:gastric cancer stem cells enolase-1 CD44 invasion signal pathway |
| 基金项目:中国医学科学院医学与健康科技创新工程（2016-I2M-3-013）； |
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| 中文摘要: |
| 摘 要：［目的］ 探讨ENO1在胃癌干细胞中的表达及其对胃癌侵袭转移的影响。［方法］ 以胃癌SNU-5模型，实时荧光定量PCR和双色细胞免疫荧光检测ENO1在其分选的CD44+细胞中的表达情况。运用慢病毒载体稳定干扰SUN-5中ENO1的表达，并用实时荧光定量PCR和Western blot检验干扰效率。流式细胞术分选稳定干扰ENO1的SNU-5中CD44+细胞后（shRNA-ENO1），实时荧光定量PCR检测其Oct-4、Sox 2以及干性相关基因Nanog的表达，同时分别进行细胞成球实验、体外侵袭与体内致瘤的胃癌干细胞生物学特性研究，Western blot检测ENO1对胃癌干细胞侵袭转移影响的相关分子机制。［结果］ SNU-5的CD44+细胞中ENO1基因和蛋白表达明显高于CD44-细胞，并建立稳定干扰ENO1的SUN-5。shRNA-ENO1细胞中干性基因Oct-4、Sox 2和Nanog中mRNA的表达显著低于PLV-Ctr细胞（P<0.05）。与PLV-Ctr细胞相比较，shRNA-ENO1细胞的自我更新能力、侵袭能力和肿瘤重量显著降低。Western blot检测shRNA-ENO1细胞中Vimentin、Snail和N-cadherin下调表达，同时E-cadherin上调表达，并伴随AKT和PI3K的磷酸化水平降低，提示ENO1的作用可能通过PI3K/AKT信号通路激活。［结论］ ENO1在胃癌干细胞中高表达，其在调控胃癌侵袭转移能力中发挥重要作用。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the expression of enolase-1 (ENO1) in gastric cancer stem cells (GCSCs) and its relation to the invasion and metastasis of gastric cancer. ［Methods］ CD44+ cells were sorted from human gastric cancer SNU-5 cells，the expression of ENO1 mRNA and protein was detected by RT-PCR and two-color immunofluorescence method. SUN-5 cells and sorted CD44+ cells were transfected with short hairpin RNA (shRNA) to knock down the expression of ENO1. The expression of Oct-4，Sox 2 and stemness-related gene Nanog was detected by RT-PCR，and the biological characteristics of GCSCs were examined by sphere colony formation assay，invasion assay and in vivo tumorigenicity. ［Results］ The expression of ENO1 mRNA and protein in CD44+ cells from SNU-5 was significantly higher than that in CD44- cells. The mRNA expression of Oct-4，Sox 2 and Nanog from shRNA-ENO1 cells was significantly lower than that of control PLV-Ctr cells (P<0.05). Compared with the PLV-Ctr cells，the shRNA-ENO1 cells showed significantly decreased self-renewal capacity and invasion ability，and the tumor growth in vivo was inhibited. Western blot showed that Vimentin，snail and N-cadherin was up-regulated in shRNA-ENO1 cells，while E-cadherin was down-regulated and the phosphorylation of AKT and PI3K was decreased，suggesting that ENO1 may be activated by AKT/PI3K signaling pathway. ［Conclusion］ ENO1 is highly expressed in GCSCs，which may be involved in the invasion and metastasis of gastric cancer. |
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