BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact mo...BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.展开更多
文摘目的 探讨miR-383在单钠尿酸盐(monosodium urate, MSU)诱导的非感染性类病原炎症反应中的调控作用,重点评估其在NLRP3炎性体活性调节及炎症自限性维持中的分子机制,为DAMPs介导的无菌性炎症调控提供理论基础。方法 构建miR-383基因敲除(miR-383-/-)及野生型(WT)小鼠的急性痛风性关节炎模型,分别于MSU注射后0、6、12、24、48 h测定足掌厚度。建立腹腔炎症模型并于刺激后6 h和12 h收集腹腔灌洗液,采用流式细胞术分析Ly6G+CD11b+中性粒细胞与F4/80+CD11b+巨噬细胞构成比,ELISA检测IL-1β与IL-6水平。另分离骨髓源性巨噬细胞(BMDMs),于MSU刺激(100μg/mL)后0、4、8 h收集RNA,利用RT-qPCR检测NLRP3、ASC、caspase-1与IL-1β mRNA表达水平。结果 miR-383-/-小鼠在MSU注射后6 h、12 h、24 h的足掌厚度较WT组显著增高(2.81±0.17 mm vs. 2.37±0.14 mm, 3.08±0.21 mm vs. 2.52±0.19 mm, 2.67±0.15 mm vs. 2.29±0.12 mm,均P<0.01)。12 h时腹腔中Ly6G+CD11b+中性粒细胞比例升高至72.34%±4.26%,显著高于WT组的60.91%±3.74%(P<0.01),F4/80+CD11b+巨噬细胞占比亦显著上升(18.46%±2.81%vs. 12.73%±2.57%,P<0.01)。ELISA结果显示miR-383-/-组IL-1β和IL-6峰值分别达641.5±43.6 pg/mL和984.2±51.8 pg/mL,均显著高于WT组(分别为438.7±39.4 pg/mL和732.4±44.9 pg/mL,均P<0.01)。BMDMs中,miR-383-/-组在刺激4 h时ASC mRNA表达为WT组的2.91倍(P<0.01),caspase-1和IL-1β表达分别升高至2.26倍(P<0.01)与3.42倍(P<0.01),NLRP3表达差异无统计学意义(P=0.117)。结论 miR-383通过负向调控ASC表达,从而限制NLRP3炎性体的激活及促炎因子的过度释放。其缺失导致炎症表型加重、免疫细胞过度募集及IL-1β、IL-6水平显著升高,提示miR-383在DAMPs介导的无菌性炎症应答中发挥关键负调控功能。miR-383-ASC信号轴的识别不仅丰富了炎性小体调控网络,也为痛风等炎症性疾病的分子干预提供潜在靶点。
基金Supported by the Haihe Laboratory of Cell Ecosystem Innovation Fund,No.22HHXBJC00001the Key Discipline Special Project of Tianjin Municipal Health Commission,No.TJWJ2022XK016.
文摘BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.
