幽门螺杆菌感染对人胃黏膜和胃癌细胞热休克蛋白70表达的影响

陶虹锦; 张少杰; 王炳智; 薛丽燕; 刘静; 王刚石

doi:10.3969/j.issn.2095-5227.2022.03.019

幽门螺杆菌感染对人胃黏膜和胃癌细胞热休克蛋白70表达的影响

Effects of Helicobacter pylori infection on expression of heat shock protein 70 in human gastric mucosa and gastric cancer cells

摘要

摘要:
背景热休克蛋白70(heat shock protein 70，HSP70)是细胞在不利应激下产生的保护蛋白，幽门螺杆菌(Helicobacter pylori，Hp)感染可能影响胃黏膜细胞HSP70的表达。
目的研究Hp感染对人胃黏膜和胃癌细胞HSP70表达的影响，探讨HSP70在人胃黏膜组织的表达特征。
方法将人胃黏膜细胞Ges-1和胃癌细胞BGC-823分别与Hp ATCC 26695共培养，采用Western-blot及RT-PCR/qPCR方法检测HSP70蛋白及mRNA在细胞水平表达的变化。利用免疫组织化学染色(IHC)检测人胃黏膜组织的Hp感染和HSP70表达，观察HSP70蛋白在胃癌组织和正常胃腺体组织的分布规律。
结果 Hp感染的Ges-1细胞和BGC-823细胞均出现了HSP70 mRNA表达一过性下调，Ges-1细胞和BGC-823细胞分别在与Hp共培养的12 h和24 h下调最为显著，至48 h则mRNA表达水平回升；而在蛋白水平，Ges-1细胞和BGC-823细胞内HSP70蛋白的表达在48 h内逐渐下降。IHC显示，组织切片中Hp长期感染的人胃黏膜腺体HSP70蛋白表达无明显下降；与正常胃腺体组织相比，胃癌组织的HSP70蛋白高表达；在正常胃腺体中，胃泌酸腺体和腺颈部增殖带的细胞也表现出一定程度的HSP70表达，而分化成熟的胃小凹上皮细胞无HSP70表达。
结论 Hp感染导致的正常胃黏膜细胞和胃癌细胞HSP70表达下调具有时间规律性，其中在体外试验中Ges-1细胞和BGC-823细胞的HSP70 mRNA水平在12 h和24 h下调到最低点，随后开始逐渐回升；HSP70蛋白表达在48 h内逐渐下调，而在体内长期感染Hp的胃黏膜组织中表达水平无下调趋势。因此，Hp感染导致的HSP70表达下调呈现出一过性的特点。

Abstract:
Background Heat shock protein 70 (HSP70) is a protective protein produced by cells under adverse stress, and its expression in gastric mucosa cells can be affected by Helicobacter pylori (Hp) infection.
Objective To investigate the effect of H.pylori infection on the expression of HSP70 in human gastric mucosa and gastric cancer cells, and explore the expression characteristics of HSP70 in human gastric mucosa.
Methods Human gastric mucosa cells (Ges-1) and gastric cancer cells (BGC-823) were co-cultured with Hp ATCC 26695 respectively. The expression of HSP70 protein and mRNA were detected by Western-blot and RT-PCR/qPCR. Immunohistochemical staining (IHC) was used to detect Hp infection and HSP70 expression in gastric mucosa, and the distribution patterns of HSP70 protein in gastric cancer and normal gastric gland tissues were also analyzed.
Results HSP70 mRNA levels in both Ges-1 and BGC-823 cells co-cultured with Hp were downregulated transiently. The downregulation was most significant at 12 h in Ges-1, and at 24 h in BGC-823 cells. However, their mRNA expression levels increased at 48 h, while the expression level of HSP70 protein in Ges-1 cells and BGC-823 cells decreased gradually within 48 h. IHC showed no significant decrease in HSP70 protein expression in long-term Hp infected human gastric mucosa glands in tissue sections. Compared with normal gastric glands, HSP70 protein was highly expressed in gastric cancer tissues. HSP70 expression was also found in the oxyntic glands and proliferative cells in the normal gastric mucosa, but not in the mature gastric pit epithelial cells.
Conclusion The downregulation of HSP70 expression in normal gastric mucosa cells and gastric cancer cells caused by Hp infection exhibits a time dependent rhythm. The HSP70 mRNA levels of Ges-1 and BGC-823 cells decrease to the lowest point at 12 h and 24 h in vitro, and then begin to rise gradually. The expression of HSP70 protein is down-regulated gradually within 48 h, but there is no down-regulation trend in gastric mucosa tissue with long-term infection in vivo. Therefore, the down-regulation of HSP70 expression caused by Hp infection shows transient characteristics.

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