长链非编码RNA浆细胞瘤多样异位基因1对幽门螺旋杆菌感染胃上皮细胞系引起的炎症反应和细胞迁移的影响  

Effects of Long Non-coding RNA Plasmacytoma Variant Translocation 1 Gene on Inflammatory Response and Cell Migration in Helicobacter Pylori Infected Gastric Epithelial Cell Line

作　　者：竟晓慧 李凌雪 韩涛涛 史娟[1] JING Xiaohui;LI Lingxue;HAN Taotao;SHI Juan(National Laboratory of Medical Molecular Biology,Institute of Basic Medical Sciences,CAMS and PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所,北京协和医学院基础学院,医学分子生物学国家重点实验室,北京100005

出　　处：《中国医学科学院学报》2020年第2期228-235,共8页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(81572755)。

摘　　要：目的研究长链非编码RNA浆细胞瘤多样异位基因1(PVT1)在幽门螺旋杆菌(HP)感染相关胃癌中的作用。方法在HP感染的胃黏膜正常上皮细胞(GES-1)中,用实时定量PCR(qRT-PCR)检测PVT1的表达;通过瞬时转染小干扰RNA,在胃癌细胞系SGC-7901中敲低PVT1,然后与HP共培养,用qRT-PCR检测炎症相关细胞因子肿瘤坏死因子α(TNF-α)、白介素(IL)-1β、IL-6及IL-8的表达情况;在敲低PVT1后,通过细胞划痕实验,检测PVT1对胃癌细胞增殖及迁移能力的影响。RNA下拉联合质谱分析技术检测能够与PVT1相互结合的蛋白,并用RNA下拉实验联合Western blot验证质谱分析结果的准确性。结果在HP感染的胃黏膜正常上皮细胞GES-1中,PVT1明显上调(t=7.160,P=0.019)。在胃癌细胞中敲低PVT1,再用HP感染,发现炎症因子TNF-α(t=3.899,P=0.011)、IL-1β(t=14.610,P=0.000)、IL-8(t=6.557,P=0.001)的表达受到明显抑制。PVT1敲低后,对胃癌细胞的增殖能力无影响,但抑制细胞的迁移能力。PVT1可能与RPS8蛋白相互作用。结论PVT1可能作为促炎因子促进胃癌细胞迁移,在HP感染相关的胃癌中发挥调节作用。Objective To investigate the mechanism of long non-coding RNA plasmacytoma variant translocation 1(PVT1)in gastric cancer caused by helicobacter pylori(HP)infection.Methods The expression of PVT1 was detected by quantitative real-time polymerase chain reaction in HP-infected normal gastric epithelial cells GES-1.Gastric cancer cell line SGC-7901 was transfected with PVT1 small interfering RNA and co-cultured with HP,and then the inflammatory cytokines such as tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6 and IL-8 were detected.After PVT1 was knocked down,the effects of PVT1 on the proliferation and migration of gastric cancer cells were examined by cell scratch assay.RNA-pulldown combined with mass spectrometry was used to detect the protein binding to PVT1,and the result of mass spectrometry was verified by RNA-pulldown combined with Western blot.Results In HP-infected normal gastric epithelial cells GES-1,quantitative real-time polymerase chain reaction showed that PVT1 was significantly up-regulated(t=7.160,P=0.019).PVT1 was knocked down in gastric cancer cells,and then infected with HP.The expressions of inflammatory factors including TNF-α(t=3.899,P=0.011),IL-1β(t=14.610,P=0.000),and IL-8(t=6.557,P=0.001)were significantly inhibited.Although PVT1 knockdown had no significant effect on the proliferation ability of gastric cancer cells,it inhibited the migration of cells.PVT1 might interact with RPS8 protein.Conclusion PVT1 may act as a pro-inflammatory factor and regulate gastric cancer caused by HP infection.

关 键 词：幽门螺旋杆菌 长链非编码RNA 浆细胞瘤多样异位基因1 炎症相关因子 胃癌 

分 类 号：Q74[生物学—分子生物学]