miR-3133 inhibits gastrointestinal cancer progression through activation of Hippo and p53 signalling pathways via multi-targets

Background

Malignant cell growth and chemoresistance, the main obstacles in treating gastrointestinal cancer (GIC), rely on the Hippo and p53 signalling pathways. However, the upstream regulatory mechanisms of these pathways remain complex and poorly understood.

Methods

Immunohistochemistry (IHC), western blot and RT-qPCR were used to analyse the expression of RNF146, miR-3133 and key components of Hippo and p53 pathway. CCK-8, colony formation, drug sensitivity assays and murine xenograft models were used to investigate the effect of RNF146 and miR-3133 in GIC. Further exploration of the upstream regulatory mechanism was performed using bioinformatics analysis, dual-luciferase reporter gene, immunoprecipitation assays and bisulfite sequencing PCR (BSP).

Results

Clinical samples, in vitro and in vivo experiments demonstrated that RNF146 exerts oncogenic effects in GIC by regulating the Hippo pathway. Bioinformatics analysis identified a novel miRNA, miR-3133, as an upstream regulatory factor of RNF146. fluorescence in situ hybridization and RT-qPCR assays revealed that miR-3133 was less expressed in gastrointestinal tumour tissues and was associated with adverse pathological features. Functional assays and animal models showed that miR-3133 promoted the proliferation and chemotherapy sensitivity of GIC cells. miR-3133 affected YAP1 protein expression by targeting RNF146, AGK and CUL4A, thus activating the Hippo pathway. miR-3133 inhibited p53 protein degradation and extended p53's half-life by targeting USP15, SPIN1. BSP experiments confirmed that miR-3133 promoter methylation is an important reason for its low expression.

Conclusion

miR-3133 inhibits GIC progression by activating the Hippo and p53 signalling pathways via multi-targets, including RNF146, thereby providing prognostic factors and valuable potential therapeutic targets for GIC.     

Cloning of cDNAs for human phosphoribosylpyrophosphate synthetases 1 and 2 and X chromosome localization of PRPS1 and PRPS2 genes

Cloned cDNAs representing the entire, homologous (80%) translated sequences of human phosphoribosylpyrophosphate synthetase (PRS) 1 and PRS 2 cDNAs were utilized as probes to localize the corresponding human PRPS1 and PRPS2 genes, previously reported to be X chromosome linked. PRPS1 and PRPS2 loci mapped to the intervals Xq22-q24 and Xp22.2-p22.3, respectively, using a combination of in situ chromosomal hybridization and human x rodent somatic cell panel genomic DNA hybridization analyses. A PRPS1-related gene or pseudogene (PRPS1L2) was also identified using in situ chromosomal hybridization at 9q33-q34. Human HPRT and PRPS1 loci are not closely linked. Despite marked cDNA and deduced amino acid sequence homology, human PRS 1 and PRS 2 isoforms are encoded by genes widely separated on the X chromosome.     

Antipurinergic Therapy Corrects the Autism-Like Features in the Poly(IC) Mouse Model

Background

Autism spectrum disorders (ASDs) are caused by both genetic and environmental factors. Mitochondria act to connect genes and environment by regulating gene-encoded metabolic networks according to changes in the chemistry of the cell and its environment. Mitochondrial ATP and other metabolites are mitokines—signaling molecules made in mitochondria—that undergo regulated release from cells to communicate cellular health and danger to neighboring cells via purinergic signaling. The role of purinergic signaling has not yet been explored in autism spectrum disorders.

Objectives and Methods

We used the maternal immune activation (MIA) mouse model of gestational poly(IC) exposure and treatment with the non-selective purinergic antagonist suramin to test the role of purinergic signaling in C57BL/6J mice.

Results

We found that antipurinergic therapy (APT) corrected 16 multisystem abnormalities that defined the ASD-like phenotype in this model. These included correction of the core social deficits and sensorimotor coordination abnormalities, prevention of cerebellar Purkinje cell loss, correction of the ultrastructural synaptic dysmorphology, and correction of the hypothermia, metabolic, mitochondrial, P2Y2 and P2X7 purinergic receptor expression, and ERK1/2 and CAMKII signal transduction abnormalities.

Conclusions

Hyperpurinergia is a fundamental and treatable feature of the multisystem abnormalities in the poly(IC) mouse model of autism spectrum disorders. Antipurinergic therapy provides a new tool for refining current concepts of pathogenesis in autism and related spectrum disorders, and represents a fresh path forward for new drug development.     

Genetic Polymorphisms at TIMP3 Are Associated with Survival of Adenocarcinoma of the Gastroesophageal Junction

The poor survival of adenocarcinomas of the gastroesophageal junction (GEJ) makes them clinically important. Discovery of host genetic factors that affect outcome may guide more individualized treatment. This study tests whether constitutional genetic variants in matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) genes are associated with outcome of GEJ adenocarcinoma. Single nucleotide polymorphisms (SNPs) at four TIMP (TIMP1-4) and three MMP genes (MMP2, MMP7 and MMP9) were genotyped in DNA samples from a prospective cohort of patients with primary adenocarcinoma of the GEJ admitted to the British Columbia Cancer Agency. Cox proportional hazards regression, with adjustment for patient, disease and treatment variables, was used to estimate the association of SNPs with survival. Genotypes for 85 samples and 48 SNPs were analyzed. Four SNPs across TIMP3, (rs130274, rs715572, rs1962223 and rs5754312) were associated with survival. Interaction analyses revealed that the survival associations with rs715572 and rs5754312 are specific and significant for 5FU+cisplatin treated patients. Sanger sequencing of the TIMP3 coding and promoter regions revealed an additional SNP, rs9862, also associated with survival. TIMP3 genetic variants are associated with survival and may be potentially useful in optimizing treatment strategies for individual patients.     

玉米花生间作对玉米光合特性及产量形成的影响

为了进一步揭示玉米花生间作体系中玉米间作产量优势的光合机理,于2010—2011年在河南科技大学试验农场研究了间作玉米功能叶的光-光响应曲线和光-CO2响应曲线特点、荧光参数、叶绿素含量与构成、干物质积累及灌浆速率。结果表明:间作提高了玉米功能叶片的叶绿素含量,改变了叶绿素构成,显著提高了净光合速率,延缓衰老;间作提高了玉米光补偿点、光饱和点、光饱和时的最大净光合速率、表观量子效率和羧化效率,显著降低了CO2补偿点;PSⅡ的实际光化学效率、PSⅡ的最大光化学效率和光化学猝灭系数变化不明显。间作明显提高玉米生育后期单株干物质,主要在于促进了籽粒的生长,显著提高玉米产量,偏土地当量(PLER-M)高于其所占面积比例的106.6%—120.3%,表现出明显的间作产量优势。这说明间作玉米产量间作优势主要来源于其生育后期净光合速率的提高,促进光合物质向籽粒的分配,净光合速率的提高是通过羧化效率和表观量子效率的提高,增强CO2的固定能力实现的,而非是光能传递、转化效率的提高。     

碎米荠超氧化物歧化酶基因家族成员的鉴定和分析

为了解碎米荠(Cardamine hirsuta)的SOD基因特征,对SOD家族成员的基因结构、染色体定位、系统进化关系进行了分析,对顺式作用元件和蛋白结构进行了预测,并利用qRT-PCR技术检测各家族成员的组织表达模式。结果表明,碎米荠基因组中共有10个SOD基因(ChSODs),包括6个Cu/Zn-SOD、3个Fe-SOD和1个Mn-SOD。编码的ChSODs蛋白有57~ 324个氨基酸,分子量为6 419.41~34 659.01 kDa,理论等电点为4.92~9.60;系统进化树分析表明,碎米荠的ChSOD与拟南芥的AtSOD的同源性较高;ChSODs在根、茎、叶中均有表达,且在叶中高表达,其中CARHR085500和CARHR256690在叶和茎中表达量较高;顺式作用元件预测表明,碎米荠SOD响应多种非生物胁迫,其中对ABA和低温胁迫较为敏感; ChSODs蛋白质的二级和三级结构具有差异性。这表明碎米荠SOD基因在抗氧化过程中发挥重要作用。     

喀斯特洞穴小型土壤节肢动物多样性

研究洞穴土壤节肢动物,有助于了解土壤节肢动物对特殊环境的响应,对于深入认识喀斯特生态过程具有重要意义。以喀斯特洞穴生态系统小型土壤节肢动物为研究对象,采用主成分分析、重复测量方差分析、相关性分析和冗余分析等方法探讨了小型土壤节肢动物与环境因子的相互作用关系。调查共获得小型土壤节肢动物2399个,隶属7纲15目121科。其中,自然林优势类群为等节跳科(Isotomidae),洞穴优势类群为奥甲螨科(Oppiidae)。PCA分析显示,洞穴与自然林小型土壤节肢动物群落组成差异明显。洞穴小型土壤节肢动物类群数、密度和Shannon-Wiener指数(H′)显著低于自然林(P<0.05),自然林小型土壤节肢动物类群数、密度和Shannon-Wiener指数(H′)季节差异显著(P<0.05),洞穴类群数、密度和Shannon-Wiener指数(H′)季节差异不明显。相关性分析结果表明,小型土壤节肢动物类群数和Shannon-Wiener指数(H′)与pH值、全磷和光照强度显著相关(P<0.05),密度与pH值、全磷、有机质和光照强度显著相关(P<0.05)。RDA分析表...     

Influence of number and map distribution of AFLP markers on similarity estimates in carrot

When genetic diversity among organisms was measured with molecular markers, the question of genome coverage was currently stressed out. In order to check if well-distributed, mapped AFLP markers were more efficient in assessing varietal identification of carrot accessions than randomly chosen markers, nine closely related genotypes were analysed. A software was developed to realise 1,000 random choices of 20 to 70 mapped or unmapped markers, offering numerous genome coverages. We statistically showed that taking into account marker position does not provide a better estimation of genetic distances. Moreover, in the case of carrot, we concluded that 60 AFLP markers offer the best compromise between the level of precision and minimal expense.