| 张霹雲,刘梁英,万晓强,等.lncRNA-PVT1通过Hp/CagA介导的胃黏膜屏障损伤促进胃癌的发病机制研究[J].肿瘤学杂志,2022,28(10):841-846. |
| lncRNA-PVT1通过Hp/CagA介导的胃黏膜屏障损伤促进胃癌的发病机制研究 |
| lncRNA Promotes the Pathogenesis of Gastric Cancer Through Hp/CagA-mediated Gastric Mucosal Barrier Damage |
| 投稿时间：2022-02-21 |
| DOI：10.11735/j.issn.1671-170X.2022.10.B007 |
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| 中文关键词: 胃癌 lncRNA-PVT1 Hp/CagA 胃黏膜屏障 |
| 英文关键词:gastric cancer lncRNA-PVT1 Hp/CagA gastric mucosal barrier |
| 基金项目:重庆市技术创新与应用发展专项面上项目(cstc2019iscx-msxmX0202) |
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| 中文摘要: |
| 摘 要：［目的］ 探讨lncRNA-PVT1通过Hp/CagA介导的胃黏膜屏障损伤促进胃癌的发病机制。［方法］ 检测lncRNA-PVT1在CagA阳性幽门螺杆菌菌株（Hp-CagA+）和CagA基因敲除突变体（Hp-ΔCagA-）感染的人胃癌上皮细胞系AGS中的表达。过表达PVT1，分为对照组和PVT1组。qPCR和荧光原位杂交技术测定lncRNA-PVT1的表达，CCK8和流式细胞术检测细胞活力及凋亡率。检测细胞中活性氧（ROS）水平，透射电子显微镜（TEM）观察细胞中线粒体的超微结构；Western blot检测两组中细胞周期蛋白CDK4和cyclinD1，凋亡蛋白BAX、Bcl-2和裂解的caspase3，线粒体裂变相关蛋白DRP1和MFN2，紧密连接蛋白（occludin，claudin-1和ZO-1）的表达。 ［结果］ lncRNA-PVT1在Hp-CagA+感染AGS细胞中表达增加。过表达PVT1后增加Hp-CagA+感染AGS细胞中增殖，降低了其凋亡，调节Hp-CagA+感染AGS细胞中CDK4（1.22±0.02 vs 1.65±0.03）和cyclinD1（1.16±0.05 vs 1.53±0.01）的表达，增加了ROS生成的增加和线粒体膜电位（MMP）而升高了细胞的比例，增加了线粒体脊的肿胀畸形，降低了DRP1的磷酸化和MFN2（1.16±0.06 vs 0.59±0.03）和BAX（1.05±0.06 vs 1.40±0.03）的表达，升高了Bcl-2（1.66±0.03 vs 1.24±0.02）和裂解的caspase3（1.03±0.06 vs 1.71±0.02）的表达，增加了氧化应激反应诱导的线粒体功能障碍。过表达PVT1通过降低紧密连接蛋白occludin（4.13±0.05 vs 1.31±0.02）、 claudin-1（5.03±0.06 vs 1.33±0.03）和ZO-1（1.48±0.04 vs 0.60±0.01）表达增加了细胞屏障。 ［结论］ lncRNA-PVT1通过Hp/CagA介导的胃黏膜屏障损伤促进胃癌的发病，促进肿瘤形成和发展。 |
| 英文摘要: |
| Abstract：［Objective］ To explore the effect of lncRNA-PVT1 on pathogenesis of gastric cancer and its relation with Hp/CagA-mediated gastric mucosal barrier damage. ［Methods］ Human gastric cancer epithelial AGS cells were infected with CagA-positive Helicobacter pylori strain(Hp-CagA+) or CagA gene knockout mutant Hp strain(Hp-ΔCagA-)， respectively. The expression of lncRNA-PVT1 in AGS cells was detected. The expression of lncRNA-PVT1 was measured by qPCR and fluorescence in situ hybridization techniques. Cell viability and apoptosis rate were detected by CCK8 and flow cytometry. The level of reactive oxygen species(ROS) and transmission electron microscopy(TEM) were used to observe the ultrastructure of mitochondria in cells. The expressions of cyclins CDK4 and cyclinD1， apoptosis proteins BAX， Bcl-2 and cleaved caspase3 were measured by Western blot， mitochondrial fission-related proteins DRP1 and MFN2 were used to detect the expression of tight junction proteins occludin， claudin-1 and ZO-1. ［Results］ The expression of lncRNA-PVT1 increased in Hp-CagA+ infected AGS cells. Over expression of lncRNA-PVT1 in Hp-CagA+ infected AGS cells increased cell proliferation， reduces their apoptosis， increased the expression of CDK4(1.22±0.02 vs 1.65±0.03) and cyclinD1(1.16±0.05 vs 1.53±0.01)， increased the generation of ROS and the mitochondrial membrane potential(MMP)， increased the swelling deformity of mitochondrial ridges， decreased the phosphorylation of DRP1 and the expression of MFN2（1.16±0.06 vs 0.59±0.03） and BAX（1.05±0.06 vs 1.40±0.03）， increased the expression of Bcl-2（1.66±0.03 vs 1.24±0.02） and cleaved caspase3（1.03±0.06 vs 1.71±0.02）， and increased the mitochondrial dysfunction induced by oxidative stress. In addition， overexpression of PVT1 increased the cell barrier by reducing the tight junction proteins occludin（4.13±0.05 vs 1.31±0.02）， claudin-1（5.03±0.06 vs 1.33±0.03） and ZO-1（1.48±0.04 vs 0.60±0.01）. ［Conclusion］ The study indicates that lncRNA-PVT1 may promote the pathogenesis and development of gastric cancer through Hp/CagA-mediated gastric mucosal barrier damage. |
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