LncRNA HIF1A-AS2 accelerates malignant phenotypes of renal carcinoma by modulating miR-30a-5p/SOX4 axis as a ceRNA

Objective: Several reports have proposed that lncRNAs, as potential biomarkers, participate in the progression and growth of malignant tumors. HIF1A-AS2 is a novel lncRNA and potential biomarker, involved in the genesis and development of carcinomas. However, the molecular mechanism of HIF1A-AS2 in renal carcinoma is unclear.Methods:The relative expression levels of HIF1A-AS2 and miR-30a-5p were detected using RT-qPCR in renal carcinoma tissues and cell lines. Using loss-of-function and overexpression, the biological effects of HIF1A-AS2 and miR-30a-5p in kidney carcinoma progression were characterized. Dual luciferase reporter gene analysis and Western blot were used to detect the potential mechanism of HIF1A-AS2 in renal carcinomas.Results:HIF1A-AS2 was upregulated in kidney carcinoma tissues when compared with para-carcinoma tissues (P < 0.05). In addition, tumor size, tumor node mestastasis stage and differentiation were identified as being closely associated with HIF1A-AS2 expression (P < 0.05). Knockdown or overexpression of HIF1A-AS2 either restrained or promoted the malignant phenotype and WNT/β-catenin signaling in renal carcinoma cells (P < 0.05). MiR-30a-5p was downregulated in renal cancers and partially reversed HIF1A-AS2 functions in malignant renal tumor cells. HIF1A-AS2 acted as a microRNA sponge that actively regulated the relative expression of SOX4 in sponging miR-30a-5p and subsequently increased the malignant phenotypes of renal carcinomas. HIF1A-AS2 showed a carcinogenic effect and miR-30a-5p acted as an antagonist of the anti-oncogene effects in the pathogenesis of renal carcinomas.Conclusions:The HIF1A-AS2-miR-30a-5p-SOX4 axis was associated with the malignant progression and development of renal carcinoma. The relative expression of HIF1A-AS2 was negatively correlated with the expression of miR-30a-5p, and was closely correlated with SOX4 mRNA levels in renal cancers.     

Diagnostic exome sequencing of syndromic epilepsy patients in clinical practice

Although genetic revolution of recent years has vastly expanded a list of genes implicated in epilepsies, complex architecture of epilepsy genetics is still largely unknown, consequently, universally accepted workflows for epilepsy genetic testing in a clinical practice are missing. We present a comprehensive NGS‐based diagnostic approach addressing both the clinical and genetic heterogeneity of disorders involving epilepsy or seizures. A bioinformatic panel of 862 epilepsy‐ or seizure‐associated genes was applied to Mendeliome (4813 genes) or whole‐exome sequencing data as a first stage, while the second stage included untargeted variant interpretation. Eighty‐six consecutive patients with epilepsy or seizures associated with neurodevelopmental disorders and/or congenital malformations were investigated. Of the 86 probands, 42 harbored pathogenic and likely pathogenic variants, giving a diagnostic yield of 49%. Two patients were diagnosed with pathogenic copy number variations and 2 had causative mitochondrial DNA variants. Eleven patients (13%) were diagnosed with diseases with specific treatments. Besides, genomic approach in diagnostics had multiple additional benefits due to mostly non‐specific, overlapping, not full‐blown phenotypes and abilities to diagnose novel and ultra rare epilepsy‐associated diseases. Likely pathogenic variants were identified in SOX5 gene, not previously associated with epilepsy, and UBA5, a recently associated with epilepsy gene.     

SOX7 interferes with β‐catenin activity to promote neuronal apoptosis

SOX7 mediates various developmental processes. However, its role in neuronal apoptosis remains unclear. In the present study, we investigated the expression pattern and role of SOX7 in potassium deprivation‐induced rat cerebellar granule neuron apoptosis. Our results showed that both mRNA and protein levels of SOX7 were upregulated when potassium was deprived. SOX7 overexpression promoted neuronal apoptosis, whereas knockdown of SOX7 protected neurons against apoptosis. Moreover, we found that β‐catenin activity was suppressed during apoptosis and that β‐catenin inhibition was crucial for potassium deprivation‐induced neuronal apoptosis. This suppression was mediated by an interaction between SOX7 and β‐catenin but not by protein degradation. Lastly, we showed that β‐catenin inhibition mediated the pro‐apoptotic effect of SOX7. Together, our findings demonstrated that SOX7 interfered with β‐catenin activity to promote neuronal apoptosis, which acted as a novel signaling mechanism in neuronal cell death.     

Hypotrichosis‐lymphedema‐telangiectasia‐renal defect associated with a truncating mutation in the SOX18 gene

SOX18 mutations in humans are associated with both recessive and dominant hypotrichosis–lymphedema–telangiectasia syndrome (HLTS). We report two families with affected children carrying a SOX18 mutation: a living patient and his stillborn brother from Canada and a Belgian patient. The two living patients were diagnosed with HLTS and DNA analysis for the SOX18 gene showed that both had the identical heterozygous C > A transversion, resulting in a pre‐mature truncation of the protein, lacking the transactivation domain. Both living patients developed renal failure with severe hypertension in childhood for which both underwent renal transplantation. To our best knowledge this is the first report of renal failure associated with heterozygous mutations in the SOX18 gene. We conclude that this specific mutation results in a new, autosomal dominant condition and propose the acronym HLT‐renal defect syndrome for HLTRS.     

Regulation of SOX10 stability via ubiquitination-mediated degradation by Fbxw7α modulates melanoma cell migration

Dysregulation of SOX10 was reported to be correlated with the progression of multiple cancer types, including melanocytic tumors and tumors of the nervous system. However, the mechanisms by which SOX10 is dysregulated in these tumors are poorly understood. In this study, we report that SOX10 is a direct substrate of Fbxw7α E3 ubiquitin ligase, a tumor suppressor in multiple cancers. Fbxw7α promotes SOX10 ubiquitination-mediated turnover through CPD domain of SOX10. Besides, GSK3β phosphorylates SOX10 at CPD domain and facilitates Fbxw7α-mediated SOX10 degradation. Moreover, SOX10 protein levels were inversely correlated with Fbxw7α in melanoma cells, and modulation of Fbxw7α levels regulated the expression of SOX10 and its downstream gene MIA. More importantly, SOX10 reversed Fbxw7α-mediated suppression of melanoma cell migration. This study provides evidence that the tumor suppressor Fbxw7α is the E3 ubiquitin ligase responsible for the degradation of SOX10, and suggests that reduced Fbxw7α might contribute to the upregulation of SOX10 in melanoma cells.     

不同波段电磁辐射致大鼠Sertoli细胞SOX9和WT1表达的影响

目的 探讨不同波段电磁辐射对大鼠睾丸支持细胞(Sertoli细胞)性别决定相关基因SOX9和WT1表达的影响。方法 原代分离3周Wistar大鼠的Sertoli细胞,应用WT1免疫细胞化学染色鉴定细胞纯度;Sertoli细胞经平均功率密度为100 mW/cm²的S波段微波和X波段微波及场强6×10⁴ V/m的电磁脉冲辐射4 min。应用实时RT-PCR和Western blot方法,检测SOX9、WT1的mRNA和蛋白质的变化。结果 3种波段电磁辐射后Sertoli细胞SOX9 mRNA和蛋白表达在辐射后6和12 h及辐射后的6和24 h均见不同程度的降低(F=15.20和4.84, P<0.05;F=8.46和7.47,P<0.05);WT1 mRNA和蛋白表达在辐射后6和12 h及辐射后的24 h出现不同程度降低(F=13.46、5.08和10.26,P<0.05)。结论 3种波段电磁辐射后Sertoli细胞性别决定相关基因(SOX9、WT1)表达呈不同程度的降低。Sertoli细胞SOX9、WT1的变化可能参与了电磁辐射致生殖危害的病理生理过程。     

应用引物原位标记技术快速检测SOX2基因

目的 应用引物原位标记(primed in situ labeling,PRINS)代替荧光原位杂交技术对SOX2基因进行快速检测.方法 常规制备人外周血染色体并制片,利用SOX2基因特异性引物在染色体上原位扩增包含生物素标记的探针.用酪胺信号放大(tyramide signal amplification,TSA)生物素系统处理经标记的探针.最后用链霉亲和素-德克萨斯红(streptavidin-Texas red)对生物素信号进行荧光染色,用DAPI染料对染色体进行复染.作为对照,MCF-10F细胞用慢病毒载体转染导人SOX2基因,使用相同的方法检测结合位点.结果 VideoTesT-FISH软件分析提示,实验组3号染色体长臂端有特异红色荧光信号表达,空白组与未经TSA信号处理的对照组则无特异性的红色荧光信号.经过转染的MCF-10F细胞则表现出不同的SOX2基因整合效果.结论 PRINS技术利用高敏感度的原位PCR技术(in situ PCR)结合荧光标记的脱氧核苷酸可在细胞内原位合成形成探针,可大大减少探针的费用与检测的时间,提高检测效率.在诱导多能干细胞的鉴定与分类中具有较为广泛的应用前景.