RESUMO
OBJECTIVE: Gastric cancer is the second highest mortality tumor and the fourth most common cancer worldwide that has high aggressiveness. MicroRNA-181d (miR-181d) has been established to be a tumor suppressor, by suppressing cell proliferation, cell cycle, and promoting apoptosis in several cancers. The purpose of this study is to explore the great roles of miR-181d in gastric cancer. PATIENTS AND METHODS: The Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) and Western blot were applied to calculate the mRNA and protein levels of miR-181d and genes. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and transwell assays were utilized to measure the proliferative and invasive abilities. The Kaplan-Meier method was conducted to calculate the overall survival of gastric cancer patients. RESULTS: MiR-181d was detected to be downregulated in gastric cancer tissues and cell lines compared to the peritumoral normal tissues and normal cell line. Downregulation of miR-181d predicted poor prognosis of gastric cancer patients. Cylindromatosis gene (CYLD) was overexpressed in gastric cancer tissues, which was confirmed to be a target gene of miR-181d in gastric cancer cell line HGC-27. Moreover, miR-181d inhibited the proliferation through CYLD/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway and inhibited the invasion-mediated epithelial-mesenchymal transition (EMT) in HGC-27 cells. In addition, overexpression of miR-181d suppressed tumor growth and xenograft tumorigenesis of HGC-27 cells in vivo. CONCLUSIONS: MiR-181d functioned as a tumor suppressor by inhibiting the proliferation via PI3K/AKT pathway in vitro and in vivo and inhibiting invasion-mediated epithelial-mesenchymal transition (EMT) by targeting CYLD in gastric cancer. The newly identified miR-181d/CYLD axis provides novel insight into the pathogenesis of gastric cancer.
Assuntos
Proliferação de Células , MicroRNAs/metabolismo , Transdução de Sinais , Neoplasias Gástricas/patologia , Regiões 3' não Traduzidas , Animais , Antagomirs/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Enzima Desubiquitinante CYLD/química , Enzima Desubiquitinante CYLD/genética , Enzima Desubiquitinante CYLD/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Fosfatidilinositol 3-Quinase/metabolismo , Prognóstico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/mortalidade , Taxa de Sobrevida , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: Successful cryopreservation of gametophytic material of bryophytes requires pretreatment with sucrose or abscisic acid. Compared to gametophyte materials, spore and gemmae cryopreservation may be more efficient, simple and stable systems for storing large amounts of genetic diversity of bryophytes within a small space. However, there has still been no attempt at cryopreserving bryophyte gemmae. OBJECTIVE: The aim of this study is to determine whether bryophyte gemmae with differing levels of desiccation tolerance could survive and germinate after cryopreservation without prior encapsulation and pretreatment. MATERIALS AND METHODS: Gemmae of Marchantia polymorpha L. were dried with silica gel for different times and then rapidly cooled in liquid nitrogen. RESULTS: The germination level of fresh gemmae was 95 % After 3 h predrying and 1 d in LN, germination was 68 % and was still up to 59 % after storage for 75 days. CONCLUSION: We conclude that the natural desiccation tolerance of bryophyte gemmae permits cryopreservation without prior pretreatment other than drying.
