| 曾茹,毛山山.miR-7对Hp诱导胃上皮细胞自噬及EMT的影响.[J].中南医学科学杂志.,2024,(2):177-181. |
| miR-7对Hp诱导胃上皮细胞自噬及EMT的影响 |
| Effects of miR-7 on Hp-induced autophagy and EMT in gastric epithelial cells |
| 投稿时间：2023-05-23 修订日期：2023-09-11 |
| DOI：10.15972/j.cnki.43-1509/r.2024.02.005 |
| 中文关键词: 微小RNA-7 幽门螺杆菌 胃上皮细胞GES-1 自噬 [ |
| 英文关键词:micro RNA-7 helicobacter pylori gastric epithelial cell GES-1 autophagy |
| 基金项目:海南省自然科学基金高层次人才项目(822RC862) |
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| 中文摘要: |
| 目的探讨微小RNA-7(miR-7)对幽门螺杆菌(Hp)诱导的胃上皮细胞(GES-1)自噬及上皮细胞间质转化(EMT)的影响。 方法体外将Hp悉尼株(SS1)与GES-1细胞共培养建立GES-1细胞转化模型；实验分为空白对照组(NC组)、Hp组、miR-7模拟物(mimics)组、Hp+miR-7 mimics组。qRT-PCR验证转染效果；MTT法、划痕实验、Transwell实验检测各组GES-1细胞增殖能力、迁移能力和侵袭能力；透射电镜下观察细胞自噬体；免疫印迹法检测各组GES-1细胞自噬相关蛋白[RM-9106微管相关蛋白1轻链3-Ⅰ(LC3-Ⅰ)、LC3-Ⅱ]及EMT相关蛋白[E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)]表达情况。 结果与NC组比较,Hp组GES-1转化细胞增殖抑制率、p62蛋白、E-Cadherin蛋白水平降低(P＜0.05),迁移率、侵袭数、自噬体数量及自噬泡/胞质总面积、LC3-Ⅱ/LC3-Ⅰ比值、N-Cadherin和Vimentin蛋白水平均升高(P＜0.05),miR-7 mimics组GES-1细胞增殖抑制率、p62蛋白、E-Cadherin蛋白水平升高(P＜0.05),迁移率、侵袭数、自噬体数量及自噬泡/胞质总面积、LC3-Ⅱ/LC3-Ⅰ比值、N-Cadherin和Vimentin蛋白水平均降低(P＜0.05)；与miR-7 mimics组比较,Hp+miR-7 mimics组转化细胞增殖抑制率、p62蛋白、E-Cadherin蛋白水平降低(P＜0.05),迁移率、侵袭数、自噬体数量及自噬泡/胞质总面积、自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值、N-Cadherin和Vimentin蛋白水平均升高(P＜0.05)。 结论miR-7下调Hp诱导的GES-1细胞自噬,降低细胞侵袭及迁移能力,抑制EMT。 |
| 英文摘要: |
| AimTo investigate the effect of microRNA-7 (miR-7) on Helicobacter pylori (Hp)-induced autophagy and epithelial mesenchymal transition (EMT) in gastric epithelial cells (GES-1). MethodsThe Hp Sydney strain (SS1) was co cultured with GES-1 cells in vitro to establish a GES-1 cell transformation model. The experiment was divided into blank control group (NC group), Hp group, miR-7 mimics group, and Hp+miR-7 mimics group. The transfection effect was verified by qRT-PCR. MTT assay, scratch assay and Transwell assay were used to detect the proliferation, migration and invasion of GES-1 cells in each group. Western blotting was used to detect the expression of autophagy related proteins (RM-9106 microtubule related protein 1 light chain 3-Ⅰ (LC3-Ⅰ), LC3-Ⅱ) and epithelial mesenchymal transformation (EMT) related proteins (E-cadherin, N-cadherin, Vimentin) in GES-1 cells in each group. ResultsCompared with the NC group, the Hp group showed a decrease in the proliferation inhibition rate, p62 protein, and E-Cadherin protein levels of GES-1 transformed cells (P＜0.05), while the migration rate, invasion number, number of autophagosomes, total area of autophagic vesicles/cytoplasm, ratio of LC3-Ⅱ/LC3-Ⅰ, N-Cadherin and Vimentin protein levels increased (P＜0.05). The miR-7 mimics group also showed an increase in the proliferation inhibition rate, p62 protein, and E-Cadherin protein levels of GES-1 cells (P＜0.05), and the migration rate The number of invasions, the number of autophagosomes, the total area of autophagic vesicles/cytoplasm, the ratio of LC3-Ⅱ/LC3-Ⅰ, and the levels of N-Cadherin and Vimentin proteins all decreased (P＜0.05); Compared with the miR-7 mimics group, the Hp+miR-7 mimics group showed a decrease in the inhibition rate of transformed cell proliferation, p62 protein levels, and E-Cadherin protein levels (P＜0.05), while the migration rate, invasion number, number of autophagosomes, total area of autophagic vesicles/cytoplasm, ratio of autophagy related protein LC3-Ⅱ/LC3-Ⅰ, and levels of N-Cadherin and Vimentin proteins all increased (P＜0.05). ConclusionmiR-7 down-regulates Hp-induced GES-1 autophagy, reduces cell invasion and migration, and inhibits EMT. |
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