HBV-ACLF患者血清miR-122和 HMGB1水平及其与病情、预后的关系

[摘要]　目的　探究HBV相关慢加急性肝衰竭（HBV-related acute-on-chronic liver failure, HBV-ACLF）患者血清中微小核糖核酸（microRNA, miR）-122和高迁移率族蛋白1（high-mobility group box-B1, HMGB1）水平及其与病情、预后的关系。方法　回顾性分析2016年1月—2018年1月我院收治的120例HBV-ACLF患者的一般及临床资料。根据临床结局，将患者分为存活组（53例）和死亡组（67例）。比较2组患者的一般资料、实验室检查指标及血清miR-122、HMGB1水平。多因素Logistic回归分析影响患者预后的因素。Pearson检验分析miR-122、HMGB1水平分别与TBIL、PA、终末期肝病评分模型（the model of end-stage liver disease score, MELD）评分的相关性。ROC曲线分析miR-122和HMGB1水平对患者的死亡预测价值，获得最佳临界值。根据临界值将患者分为A组、B组和C组，用Kaplan-Meier法绘制生存曲线，比较3组患者在3年随访期间的生存率。结果　存活组和死亡组患者的年龄、身体质量指数、并发症、病情分期、MELD评分、ALB、球蛋白、TBIL、ALT、AST、LDH、PT、PTA、HBV DNA、miR-122、HMGB1相比，差异均具有统计学意义（P均＜0.05）。年龄、并发症、病情分期、MELD评分、TBIL、PT、PTA、miR-122、HMGB1均是影响患者预后的危险因素（P均＜0.05）。miR-122、HMGB1水平分别与TBIL、MELD评分呈显著正相关，与PTA呈显著负相关（P均＜0.05）。miR-122和HMGB1预测患者死亡的最佳临界值分别为31.42和14.56 μg/L。A组患者预后3年内生存率显著高于B组和C组（P均＜0.05）。结论　miR-122和HMGB1水平与HBV-ACLF患者的病情和死亡预后密切相关，可间接反映患者的病情严重程度，在HBV-ACLF的诊断及预后中具有重要价值。     

LINC00649通过miR-424-5p/IGF1R轴调控内质网应激介导的宫颈癌细胞凋亡

目的探讨LINC00649/miR-424-5p/IGF1R对内质网应激(ERs)介导的宫颈癌(CC)细胞凋亡的影响。方法从GEO数据库中获取CC相关的数据,并分析差异表达的miRNAs。利用生物信息学数据库预测miR-424-5p的上、下游靶点,将LINC00649和IGF1R纳入研究,随后双荧光素酶实验进一步验证靶向关系。qRT-PCR检测LINC00649、miR-424-5p和IGF1R在CC组织和细胞中的表达水平。CCK-8和流式细胞术分别评估CC细胞增殖和凋亡变化。Western blot检测ERs相关蛋白GRP78、CHOP和Caspase-12的表达。结果与癌旁组织和H8细胞相比,LINC00649和IGF1R在CC组织和细胞中表达上调,而miR-424-5p下调(均P<0.05)。LINC00649的异常高表达与CC患者的预后不良有关,敲减LINC00649可通过促进ERs来抑制CC细胞活力,诱导细胞凋亡(均P<0.05)。LINC00649吸附miR-424-5p上调IGF1R的表达。miR-424-5p抑制剂或过表达IGF1R均可部分逆转敲减LINC00649对CC细胞的影响(均P<0.05)。结论 LINC00649能够通过miR-424-5p/IGF1R抑制CC细胞的ERs过程进而减少细胞凋亡,提高细胞活力。     

Dynamic changes in miR-124 levels in patients with acute cerebral infarction

Objective: To investigate the changes in serum miR-124 levels in patients with acute cerebral infarction (ACI) and elucidate the underlying mechanism by a dynamic monitor.

Methods: Fifty-four patients with ACI and 51 healthy controls were included in our study. Baseline characteristics and blood samples were collected for further analysis. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the serum miR-124 levels. The dual-luciferase reporter assay was used to evaluate the effect of miR-124 on iASPP, a protein that inhibits apoptosis stimulating proteins in the p53 family.

Results: Compared with normal controls, the miR-124 levels in the ACI group rapidly decreased at phase 1 (within 24?h after ischemia) (p?<?0.001) and then gradually increased at phase 2 (48?～?72?h after ischemia) (p?<?0.001) and phase 3 (the 7th day after ischemia) (p?<?0.001). The dual-luciferase reporter assay showed that miR-124 down-regulates iASPP expression in 293T cells.

Conclusion: The miR-124 levels are down-regulated in ACI patients. The dynamic changes of miR-124 might provide a possible method for the detection of ischemic stroke.

• Highlights

• The difference in miR-124 expression levels between ACI patients and normal controls.

• Dynamic changes of miR-124 expression levels in ACI patients.

• The down-regulation of miR-124 upon iASPP expression.

     

miR-30c Impedes Glioblastoma Cell Proliferation and Migration by Targeting SOX9

miR-30c has been acknowledged as a tumor suppressor in various human cancers, such as ovarian cancer, gastric cancer, and prostate cancer. However, the role of miR-30c in glioblastoma (GBM) needs to be investigated. In our study, we found that the expression of miR-30c was significantly downregulated in GBM tissues and cell lines. We found that overexpression of miR-30c inhibited cellular proliferation of GBM cells in vitro and in vivo. More GBM cells were arrested in the G₀ phase after miR-30c overexpression. Moreover, we showed that miR-30c overexpression suppressed the migration and invasion of GBM cells. Mechanistically, we found that SOX9 was a direct target of miR-30c in GBM cells. Overexpression of miR-30c inhibited the mRNA and protein levels of SOX9 in GBM cells. Moreover, there was a negative correlation between the expression of miR-30c and SOX9 in GBM tissues. Finally, we showed that restoration of SOX9 in GBM cells reversed the proliferation, migration, and invasion of GBM cells transfected with miR-30c mimic. Collectively, our results demonstrated that miR-30c suppressed the proliferation, migration, and invasion of GBM cells via targeting SOX9.     

MicroRNA-301b-3p accelerates the growth of gastric cancer cells by targeting zinc finger and BTB domain containing 4

MicroRNAs (miRNAs) have been found to be aberrantly expressed and exert essential roles in the tumorigenesis and progression of gastric cancer (GC). miR-301b-3p has been recognized as a cancer-related miRNA in lung cancer, bladder cancer and hepatocellular carcinoma. However, the function of miR-301b-3p in GC progression and its underlying mechanism have not been studied yet. In this study, we found that miR-301b-3p expression was up-regulated in GC tissues compared to adjacent noncancerous tissues. Furthermore, the elevated levels of miR-301b-3p were detected in GC cell lines (SGC-7901, AGS, MKN-45 and MGC-803) as compared with GES-1 cells. Interestingly, GC tissues from patients with tumor size ≥ 5 cm and advanced tumor stages showed obvious higher levels of miR-301b-3p compared to matched controls. Functionally, miR-301b-3p knockdown prominently inhibited cell proliferation, and induced cell cycle arrest at G1 phase and apoptosis in MGC-803 cells. Meanwhile, ectopic expression of miR-301b-3p conversely regulated these biological behaviors of MKN-45 cells. Next, we found that miR-301b-3p knockdown increased, whereas miR-301b-3p overexpression reduced the expression of zinc finger and BTB domain containing 4 (ZBTB4) in GC cells. Accordingly, luciferase reporter assay identified ZBTB4 as a direct target of miR-301b-3p. ZBTB4 overexpression markedly restrained the growth of MGC-803 cells. More importantly, ZBTB4 silencing partially reversed miR-301b-3p knockdown-induced tumor suppressive effects on MGC-803 cells. In conclusion, we firstly revealed that miR-301-3p was highly expressed in GC and contributed to tumor progression via attenuating ZBTB4, which might provide a novel molecular-targeted strategy for GC treatment.     

FBXL3 is regulated by miRNA-4735-3p and suppresses cell proliferation and migration in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is the most common type of primary lung cancer and regarded as cancer killer. The aim of this study was to discover the detailed function and molecular mechanism of F-box and leucine rich repeat protein 3 (FBXL3) in NSCLC. In this study, the expression level of FBXL3 in NSCLC tissues and cell lines was firstly examined and identified. Moreover, the relationship between FBXL3 and the overall survival rate of NSCLC patients was analyzed by Kaplan–Meier survival curve. Functionally, MTT, colony formation assay and transwell assays were performed to determine the role of FBXL3 in regulating NSCLC cell proliferation, migration and invasion. The proliferation and migration were suppressed by overexpression of FBXL3, indicating the potential tumor suppressive role of FBXL3 in NSCLC. In addition, the dual-luciferase reporter and RNA pull-down assays revealed that miR-4735-3p was a novel upstream modulator of FBXL3. Further study showed that miR-4735-3p was upregulated in NSCLC tissues and cell lines. Finally, rescue assays and function assays revealed that miR-4735-3p exerted oncogenic function in NSCLC, and this function can be attenuated by FBXL3. Taken together, FBXL3 was regulated by miR-4735-3p and suppressed cell proliferation and invasion in non-small cell lung cancer.     

miR-34c-3p与M2型肿瘤相关巨噬细胞在三阴性乳腺癌中的表达及意义

目的:检测miR-34c-3p与M2型肿瘤相关巨噬细胞（tumor-associated macrophage,TAM)在三阴性乳腺癌(triple negative breast cancer,TNBC)中的表达，并探讨其在乳腺癌发病中的意义。方法:选择2017年2月至2018年1月于西京医院就诊的68例TNBC患者作为实验组(A组)，以同期就诊的70例乳腺纤维腺瘤患者作为对照组(B组)。采用Real-time RT-PCR 检测组织标本中miR-34c-3p的表达；流式细胞检测M2型TAM的表达；ELISA法检测血清中IL-10的表达。并进一步分析miR-34c-3p的表达与乳腺癌M2型TAM及血清IL-10水平的相关性。结果:A、B组miR-34c-3p的相对表达量分别为0.84±0.08、1.29±0.71，A组明显低于B组，差异有统计学意义(P<0.05)。与B组比较，A组M2型TAM(CD68+CD163+)细胞比例显著升高(P<0.05)。A组血清IL-10的表达［(19.69±4.93) pg/ml］较B组［(17.26±3.51) pg/ml］显著升高，差异有统计学意义(P<0.05)。TNBC组织中miR-34c-3p的表达与M2型TAM比例及血清IL-10表达均呈显著负相关(r=-0.508，r=-0.656，P<0.05)。结论:TNBC组织中miR-34c-3p的表达下调，M2型TAM比例及血清IL-10表达均显著升高，促进TNBC进展。     

胶质瘤组织中miR-218的表达及其对血管生成的作用

目的 明确miR-218在胶质瘤组织中的表达，探讨其对胶质瘤血管生成的作用及机制。方法 采用荧光实时定量PCR法和免疫印迹法检测胶质瘤组织和细胞中miR-218、P70核糖体S6激酶1（p70s6k1）的表达情况，采用基质胶塞实验和小管形成实验分别在体内外检测miR-218对新血管生成的影响。结果 21例胶质瘤组织标本中miR-218表达较7例癌旁组织标本明显下调，且表达量与WHO分级有关。过表达miR-218能显著抑制血管生成。MiR-218直接靶向p70s6k1。过表达p70s6k1能部分逆转miR-218对血管新生的抑制作用。结论 MiR-218能通过靶向p70s6k1的表达调控胶质瘤血管生成，进而影响胶质瘤的进展。     

lncRNA MALAT1靶向miR-150对口腔鳞癌细胞增殖和凋亡的影响及分子机制

目的:研究MALAT1对口腔鳞癌细胞SCC-25增殖、凋亡的影响，并探讨其机制。方法:运用qRT-PCR检测人口腔鳞癌细胞SCC-25、人永生化口腔上皮细胞HIOEC中MALAT1和miR-150的mRNA表达；将si-con组（转染si-con）、si-MALAT1组（转染si-MALAT1）、miR-150组（转染miR-150 mimics）、miR-con组（转染miR-con）、si-MALAT1+anti-miR-con组（si-MALAT1和anti-miR-con共转染）、si-MALAT1+anti-miR-150组（si-MALAT1和anti-miR-150共转染），均以脂质体法转染至SCC-25细胞；MTT法检测各组细胞的增殖；流式细胞术检测各组细胞的凋亡；双荧光素酶报告基因检测实验检测各组细胞的荧光活性。结果:与人永生化口腔上皮HIOEC细胞（Normal组）相比，口腔鳞癌SCC-25细胞（Tumor组）中MALAT1表达显著上调，miR-150显著下调（P＜0.05）；敲减MALAT1、过表达miR-150均可抑制SCC-25细胞增殖，促进凋亡；MALAT1靶向miR-150。抑制miR-150逆转了敲减MALAT1对口腔鳞癌细胞的增殖抑制和凋亡促进作用。结论:MALAT1可促进口腔鳞癌细胞增殖并抑制凋亡，其机制可能与靶向miR-150有关，可为口腔鳞癌的治疗提供新靶点。