Promoter Methylation-Mediated Silencing of β-Catenin Enhances Invasiveness of Non-Small Cell Lung Cancer and Predicts Adverse Prognosis

β-Catenin plays dual role in adhesion complex formation and the Wnt signaling pathway. Although β-catenin expression appears to be upregulated and Wnt signaling pathway is activated in the majority of cancers, its expression level seems to be lost in non-small cell lung cancer (NSCLC). We previously reported that the promoter of β-catenin was hypermethylated in two NSCLC cell lines. In the current study, we expanded our analysis for the methylation status of β-catenin promoter region and its protein expression in seven NSCLC cell lines and a series of 143 cases of primary human lung cancer with adjacent non-neoplastic tissues. Quantitative methylation specific PCR (qMSP) analysis showed methylation of β-catenin promoter region in five NSCLC cell lines, with increased β-catenin protein levels upon 5′-Aza-2′-deoxycytidine (5-aza-dC) treatment. The methylation status in SPC (methylated) and A549 (unmethylated) was confirmed by bisulfite sequencing PCR. 5-Aza-dC treatment inhibited invasiveness of SPC but not A549. Immunofluorescence analysis showed membranous β-catenin expression was lost in SPC and could be re-established by 5-aza-dC, while Wnt3a treatment led to nuclear translocation of β-catenin in both SPC and A549. Dual-luciferase assays indicated that 5-aza-dC treatment caused no significant increase in Wnt signaling activity compared with Wnt3a treatment. The effect of demethylation agent in SPC can be reversed by β-catenin depletion but not E-cadherin depletion which indicated that the methylation mediated β-catenin silencing might enhance NSCLC invasion and metastasis in an E-cadherin independent manner. Subsequent immunohistochemistry results further confirmed that β-catenin promoter hypermethylation correlated with loss of immunoreactive protein expression, positive lymph node metastasis, high TNM stage and poor prognosis. The present study implicates β-catenin promoter hypermethylation in the mechanism of epigenetic changes underlying NSCLC metastasis and progression, thus indicating the potential of β-catenin as a novel epigenetic target for the treatment of NSCLC patients.     

Pseudorabies virus tegument protein UL13 recruits RNF5 to inhibit STING-mediated antiviral immunity

Pseudorabies virus (PRV) has evolved various immune evasion mechanisms that target host antiviral immune responses. However, it is unclear whether and how PRV encoded proteins modulate the cGAS-STING axis for immune evasion. Here, we show that PRV tegument protein UL13 inhibits STING-mediated antiviral signaling via regulation of STING stability. Mechanistically, UL13 interacts with the CDN domain of STING and recruits the E3 ligase RING-finger protein 5 (RNF5) to promote K27-/K29-linked ubiquitination and degradation of STING. Consequently, deficiency of RNF5 enhances host antiviral immune responses triggered by PRV infection. In addition, mutant PRV lacking UL13 impaired in antagonism of STING-mediated production of type I IFNs and shows attenuated pathogenicity in mice. Our findings suggest that PRV UL13 functions as an antagonist of IFN signaling via a novel mechanism by targeting STING to persistently evade host antiviral responses.     

近交系小型猪线粒体基因组分析和家猪系统进化关系分析

广西巴马小型猪是长期近交培育而成的实验小型猪,遗传稳定.本研究拟通过线粒体基因组(mitochondrial DNA,mtDNA)分析来了解近交系的遗传背景,并从mtDNA角度进一步认识中国本土猪种的系统进化关系.实验设计29对特异性引物,PCR扩增和产物测序,拼接出广西巴马小型猪近交系7个世代猪的mtDNA序列;用4...     

白粉菌BGP(TG)在不同抗性草地早熟禾叶片上的侵染过程观察

【背景】草地早熟禾(Poa pratensis)白粉病是气传性病害,种植抗病品种是防治白粉病最经济、环保和有效的方法。【目的】明确禾布氏白粉菌BGP(TG)在3个不同抗性草地早熟禾叶片上的侵染过程和乳突在草地早熟禾抗白粉病中起的重要角色。【方法】利用考马斯亮蓝染色结合显微镜观察的方法,对不同抗性的3个草地早熟禾品种进行白粉菌侵染过程观察,统计分生孢子形成次生芽管的速度、吸器和乳突的形成情况,分析白粉菌对不同抗性草地早熟禾侵染过程的差异。【结果】‘探险家’草地早熟禾病症较其他2个品种更为严重,禾布氏白粉菌BGP(TG)侵染不同抗性品种的侵染过程基本一致,但‘探险家’叶片上初生吸器形成早于其他2个品种;接菌1-2 d后,在‘太行’叶表面分生孢子形成5级芽管的速度显著低于‘探险家’和‘黑杰克’;接种1d后,‘太行’草地早熟禾有效乳突形成率显著高于‘黑杰克’(P<0.05),‘黑杰克’有效乳突形成率显著高于‘探险家’(P<0.05)。【结论】明确了白粉菌在草地早熟禾叶片上的侵染过程和乳突在草地早熟禾抵御白粉菌侵入过程中起的重要作用,为草地早熟禾白粉病的预防和防治提供了理论基础和参考...     

Stearoyl-CoA Desaturase-1 dependent lipid droplets accumulation in cancer-associated fibroblasts facilitates the progression of lung cancer

Rationale: Cancer-associated fibroblasts (CAFs) are the main components in the tumor microenvironment (TME) and facilitate lung cancer progression. Studies have reported that metabolic reprogramming can regulate the function of CAFs, especially abnormal lipid metabolism. Lipid droplets (LDs) are ubiquitous organelles that store neutral lipids and have a crucial role in lipid metabolism. However, little is known about the synthesis and functions of LDs in lung CAFs.Methods: TetO-EGFR^L858R; CCSP-rtTA transgenic mouse model was used to establish a spontaneous pulmonary tumor model and investigate the accumulation of LDs in CAFs. The effect of LDs accumulation on the phenotype change of fibroblasts was estimated in vitro using mouse fibroblast cell lines. RNA sequencing, Western blotting, RT-PCR, and DNA-pull down were performed to determine the mechanism of LDs synthesis in fibroblasts.Results: We found that LDs were enriched in lung CAFs and induced the pro-tumoral phenotype of CAFs with increased expression of α-smooth muscle actin (α-SMA) and Collagen alpha-2 (I) chain (COL1A2). As the main regulator, hypoxia-inducible factor-1α (HIF-1α) was highly expressed in activated fibroblasts and increased the content of LDs. RNA-sequencing results showed that Stearoyl-CoA Desaturase1 (SCD1) was a downstream gene of HIF-1α, which upregulated the number of LDs in fibroblasts. Importantly, SCD1 inhibition reduced the growth of lung tumors, which was correlated with LDs decrease in CAFs. Analysis of human lung adenocarcinoma tissue chip revealed that CAFs with a high level of SCD1 were positively correlated with the expression of HIF-1α and poor survival in lung cancer patients.Conclusions: The HIF-1α/SCD1 axis regulates the accumulation of LDs in CAFs, which might represent a novel target for lung cancer therapy.     

Cullin 4b-RING ubiquitin ligase targets IRGM1 to regulate Wnt signaling and intestinal homeostasis

Hierarchical organization of intestine relies on the self-renewal and tightly regulated differentiation of intestinal stem cells (ISCs). Although signals like Wnt are known to sustain the continued intestinal renewal by maintaining ISCs activity and lineage commitment, molecular mechanisms underlying ISCs ‘stemness’ and supportive niche have not been well understood. Here, we found that CUL4B-RING ubiquitin ligase (CRL4B) regulates intestinal homeostasis by targeting immunity-related GTPase family M member 1 (IRGM1) for proteasomal degradation. CUL4B was mainly expressed at ISCs zone. Deletion of Cul4b led to reduced self-renewal of ISCs and a decreased lineage differentiation towards secretory progenitors through downregulated Wnt signals. Besides, Cul4b-null mice exhibited impaired Paneth cells number and structure. Mechanistically, CRL4B complex were associated with WD40 proteins and targeted IRGM1 at K270 for ubiquitination and proteosomal degradation. Impaired intestinal function caused by CUL4B deletion was rescued by down-regulation of its substrate IRGM1. Our results identified CUL4B as a novel regulator of ISCs and revealed a new 26 S proteasome degradation mechanism in intestine self-renewal and lineage commitment. Subject terms: Development, Ubiquitin ligases     

宁南山区不同乔木林地土壤干层分布及干燥化特征

以宁南山区典型坡面不同人工林地为研究对象,分层采集0~600 cm土样,分析杏树(Armeniaca vulgaris)、杨树(Populus simonii)、榆树(Ulmus pumila)、刺槐(Robinia pseudoacacia)和松树(Pinus sylvestris var.mongolica)及农田(对照)土壤含水量垂直剖面分布特征,采用干燥化指数(SID)、土壤水分相对亏缺指数(CSWDI)、干层起始深度(DSLFD)、干层厚度(DSLT)和干层内平均含水量(DSL-SWC)等指标对土壤干燥化强度和干层分布特征进行分析,利用冗余分析(RDA)确定相关环境因子对土壤干燥化和干层的影响,为黄土高原山区植被合理利用水土资源和生态恢复等提供数据支撑。结果表明:(1)杏树、杨树、榆树、刺槐、松树土壤水分均属中等变异,且0~600 cm平均土壤含水量与对照农田相比分别降低了31.17%、24.15%、23.19%、29.67%和18.35%。(2)不同人工林地干层起始深度、干层厚度及垂直剖面土壤水分分布明显不同,杏树、杨树、榆树、刺槐、松树和农田土壤干层起始深度分别为90、160、140、140、160和600 cm,干层厚度分别为510、460、480、480、460和0 cm。(3)杏树、杨树、榆树、刺槐和松树0~600 cm平均土壤干燥化指数分别为63.48%、91.88%、95.17%、73.97%和111.91%,土壤水分相对亏缺指数分别为0.68、0.59、0.52、0.63和0.41,其土壤水分干燥化强度及亏缺程度由高到低依次为杏树>刺槐>杨树>榆树>松树。(4)RDA分析结果表明,乔木类型、黏粒含量和地形条件是影响土壤干燥化的重要因素。     

基于VGPM模型和MODIS数据估算梅梁湾浮游植物初级生产力

基于MODIS影像数据反演的2009年2月份至12月份太湖梅梁湾水域表面叶绿素a、悬浮物浓度以及水温数据,结合初级生产力垂向归纳模型(Vertically Generalized Production Model:VGPM)估算获得梅梁湾2009年逐月平均日初级生产力时空分布。结果表明,梅梁湾2009年年平均日初级生产力及逐月平均日初级生产力空间分布差异显著,呈现从湾内向湾口逐渐递减的趋势。时间序列分析显示,梅梁湾初级生产力季节差异显著,夏季>秋季>春季>冬季,全年初级生产力主要集中在夏季,占47.4%。通过分析VGPM模型中几个输入参数对初级生产力的影响,发现悬浮物浓度与标准化初级生产力存在显著负冥函数关系,反映沉积物再悬浮引起的悬浮物浓度增加能降低水体初级生产力。温度对初级生产力也有一定的调控与制约,与初级生产力呈现正相关趋势,在低于21℃的温度范围内与最大光合作用速率呈现正相关。