RESUMO
Helicobacter pylori (H. pylori) infection plays a pivotal role in the development of gastric cancer (GC). However, the association between aberrant microRNAs (miRNAs/miRs) expression and H. pyloriinduced GC remains poorly understood. The present study reported that repeated infection of H. pylori caused the oncogenicity of GES1 cells in BALB/c Nude mice. miRNA sequencing revealed that both miR7 and miR153 were significantly decreased in the cytotoxinassociated gene A (CagA) positive GC tissues and this was further confirmed in a chronic infection model of GES1/HP cells. Further biological function experiments and in vivo experiments validated that miR7 and miR153 can promote apoptosis and autophagy, inhibit proliferation and inflammatory response in GES1/HP cells. All the associations between miR7/miR153 and their potential targets were revealed via bioinformatics prediction and dualluciferase reporter assay. Particularly, downregulation of both miR7 and miR153 obtained an improved sensitivity and specificity in diagnosing H. pylori (CagA+)induced GC. The present study identified that the combination of miR7 and miR153 may be regarded as novel therapeutic targets in H. pylori CagA (+)associated GC.
Assuntos
Infecções por Helicobacter , Helicobacter pylori , MicroRNAs , Neoplasias Gástricas , Animais , Camundongos , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Carcinogênese/genética , Regulação para Baixo , Infecções por Helicobacter/complicações , Infecções por Helicobacter/genética , Helicobacter pylori/genética , Camundongos Nus , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias Gástricas/metabolismo , HumanosRESUMO
BACKGROUND: Gastric cancer is heterogeneous cancer and the causes of this disease are complex. New diagnostic and therapeutic targets are urgently needed to explore. Huntingtin-associated protein 1 (HAP1) is directly related to Huntington's disease (HD). However, patients with Huntington's disease have a lower incidence of cancer. Therefore, we are committed to studying the correlation between HAP1 and gastric carcinogenesis and development. METHODS AND RESULTS: Immunohistochemical staining, western blot analysis, and RT-qPCR were conducted to explore the localization and expression of HAP1 in gastric cancer. To study the biological significance of HAP1, we overexpressed HAP1 in both MKN28 and AGS cell lines by lentivirus infection. To explore the role of HAP1 in cell proliferation, the cells counting assay, EdU incorporation assay, and colony formation assay were carried out. We performed the wound healing assay and transwell assay to study the cell migration and invasion. To further investigate whether HAP1 could regulate gastric cancer cell death during glucose deprivation, Annexin V-FITC/PI staining was performed. In our study, we elucidated that HAP1 was downregulated in gastric cancer. What's more, overexpressing HAP1 inhibited cell proliferation, cell migration and invasion, and triggered apoptosis during glucose deprivation. More importantly, the antitumor properties and mechanisms of HAP1 have been elucidated further in gastric cancer. CONCLUSIONS: Taken together, the available evidence implies that HAP1 may serve as a potential tumor suppressor, making it a significant target in preventing and treating gastric cancer. This research provides a theoretical basis for the early diagnosis, clinical targeted therapy, and prognosis evaluation of gastric cancer.
Assuntos
Doença de Huntington , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/genética , Proteínas do Tecido Nervoso/metabolismo , Doença de Huntington/metabolismo , Apoptose/genética , Morte Celular , Proliferação de Células/genética , Linhagem Celular TumoralAssuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Escarro/virologia , COVID-19 , Infecções por Coronavirus/terapia , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Faringe , Pneumonia Viral/terapia , RNA Viral/isolamento & purificação , SARS-CoV-2RESUMO
The aim of this study was to determine the biological function of hpsh4590 in Helicobacter pylori. After Hpsh4590 was expressed using a prokaryotic expression system, the cytotoxic effects and IL-8 production of Hpsh4590 were analyzed by co-culturing with GES-1 cells. Meanwhile, the antibody of rHpsh4590, produced by immunizing rabbit, was used for localization and protein interaction studies. Hpsh4590 fusion protein was expressed successfully in Escherichia coli Rosetta (DE3), and the polyclonal antibody was produced at high titers. The MTT assay showed that the inhibition ratio of GES-1 cells cultured with 0.1 µg/mL rHpsh4590 (3.02% ± 0.02%) was significantly lower than that of 20 µg/mL rHpsh4590 (57.57% ± 0.03%, p < 0.01), while DAPI staining showed the cytotoxic effects of rHpsh4590 for GES-1 cells. The up-regulation of cleaved caspase-3 and cleaved PARP was observed after GES-1 cells co-cultured with rHpsh4590 by Western blot. Co-culturing of GES-1 cells with rHpsh0459 (20 µg/mL) led to significant production of IL-8 at 12 h(1097.74 ± 212.37 pg/mL) and 24 h (1379.55 ± 209.58 pg/mL) then at 6 h(134.68 ± 14.64 pg/mL, p < 0.01). These observations suggest that the cytotoxicity of Hpsh4590 occurred in a concentration dependent manner, which is related with IL-8 secretion from gastric mucosal epithelial cells. Hpsh4590 was found localized in the membrane and the periplasm of H. pylori, interacted with zinc finger protein and methionine ABC transporter ATP-binding protein, and potentially regulates DNA uptake or transfer.
