Silencing KLF16 inhibits oral squamous cell carcinoma cell proliferation by arresting the cell cycle and inducing apoptosis

Krüppel-like factor 16 (KLF16), a member of the Krüppel-like factor (KLF) family, has been extensively investigated in multiple cancer types. However, the role of KLF16 in oral squamous cell carcinoma (OSCC) remains unknown. Thus, we conducted this study to investigate its related mechanism. KLF16 expression in OSCC cell lines was quantified by western blotting. Then, OECM1 and OC3 cells were divided into Blank, siCtrl, siKLF16#1 and siKLF16#2 groups. Subsequently, cell proliferation was detected using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assays, cell migration and invasion were detected with wound healing and Transwell assays, and cell cycle distribution and cell apoptosis were detected via flow cytometry. KLF16, p21, CDK4, Cyclin D1 and p-Rb expression was detected by western blotting. Finally, xenograft models were established in nude mice to observe the in vivo effects of KLF16 on OSCC. KLF16 protein expression was upregulated in OSCC cells. Compared to the cells in the Blank group, the OECM1 and OC3 cells in the siKLF16#1 group and siKLF16#2 group exhibited a sharp decrease in proliferation but a remarkable increase in apoptosis. Moreover, the proportion of cells in the G0/G1 phase notably increased and that in the S phase decreased, with evident decreases in cell invasion and migration. Moreover, KLF16, cyclin-dependent kinase 4 (CDK4), Cyclin D1 and p-Rb protein expression was upregulated, but p21 expression was downregulated. The mice in the siKLF16#1 and siKLF16#2 xenograft model groups exhibited slower tumour growth and smaller tumours with evident downregulation of Ki67 expression compared to the mice in the Blank group. KLF16 expression was upregulated in OSCC cells, and interfering with KLF16 led to cell cycle arrest, inhibited OSCC cell growth and promoted cell apoptosis.     

Endostar,an Antiangiogenesis Inhibitor,Combined With Chemoradiotherapy for Locally Advanced Cervical Cancer

This phase II randomized clinical trial aimed to assess the efficacy and toxicity of Endostar, an antiangiogenesis inhibitor, combined with concurrent chemoradiotherapy (CCRT) for locally advanced cervical cancer (LACC). Patients with LACC were randomly assigned to either CCRT plus Endostar (CCRT+E arm) or CCRT alone (CCRT arm). All patients received pelvic intensity-modulated radiation therapy (IMRT) and brachytherapy. Weekly cisplatin was administered concurrently with IMRT. Patients in the CCRT+E arm also received concurrent Endostar every 3 weeks for two cycles. The primary endpoint was progression-free survival (PFS) and acute toxicities. The exploratory endpoint was the impact of vascular endothelial growth factor receptor-2 (VEGFR2) expression on long-term survival. A total of 116 patients were enrolled. Patients in the CCRT+E arm and in the CCRT arm had similar acute and late toxicity profile. The 1- and 2-year PFS were 91.4% versus 82.1% and 80.8% versus 63.5% (p=0.091), respectively. The 1- and 2-year distance metastasis-free survival (DMFS) were 92.7% versus 81.1% and 86.0% versus 65.1% (p=0.031), respectively. Patients with positive VEGFR2 expression had significant longer PFS and overall survival (OS) compared with those with negative VEGFR2 expression. Patients in the CCRT+E arm had significantly longer PFS, OS, and DMFS than those in the CCRT arm when VEGFR2 expression was positive. In conclusion, CCRT plus Endostar significantly improved DMFS but not PFS over CCRT alone. The addition of Endostar could significantly improve survival for patients with positive VEGFR2 expression.     

A novel lncRNA-LINC01116 regulates tumorigenesis of glioma by targeting VEGFA

Brain glioma is the most common malignant tumor of the central nervous system, and one of the leading causes of death in patients with intracranial tumors. The clinical outcome of glioma is usually poor due to abundant vascularity, fast growth and susceptibility of invasion to normal brain tissues. Our microarray study showed that lncRNA-LINC01116 was significantly upregulated in glioma tissues and played an important role in cell proliferation, cycle, migration, invasion and angiogenesis. In addition, vascular endothelial growth factor (VEGFA) may be the major target genes in the downstream of lncRNA-LINC01116. Dual luciferase assay showed that LINC01116 and VEGFA both contained a miR-31-5p binding site, and LINC01116 could regulate the expression of VEGFA through competitive absorption of miR-31-5p. RNA immunoprecipitation indicated that LINC01116 and VEGFA were present in the miR-31-5p-RISC complex, and biotinylated miR-31-5p pull-down assay suggested that there was a competitive relationship between LINC01116 and VEGFA to bind with miR-31-5p. Collectively, our study has identified a novel lncRNA-LINC01116 and clarified the role and mechanism of LINC01116 in the tumorigenesis of glioma. LINC01116 may prove to be a potential target for the clinical diagnosis and treatment of glioma.     

同型半胱氨酸对高血压患者脑卒中复发风险的影响

目的探讨高血压合并脑卒中患者的血浆同型半胱氨酸（Hcy）水平与其他危险因素对于脑卒中复发的影响。 方法分析徐州市中心医院心内科和徐州医科大学附属医院神经外科自2012年5月至2013年12月收治的1623例高血压脑卒中患者的基线资料，中位随访4.9年，根据随访事件中是否发生脑卒中分为复发组（312例）与未复发组（1311例）。Kaplan-Meier生存分析比较不同危险因素脑卒中复发率的差异，单因素与多因素Cox回归模型分析影响脑卒中复发的独立危险因素，以及危险因素之间的交互作用。 结果复发组年龄、空腹血糖、Lg Hcy的水平，以及糖尿病、房颤的患病率均高于未复发组（P<0.05）。Kaplan-Meier生存分析显示，糖尿病、房颤、年龄≥60岁、空腹血糖≥7.0 mmol/L、Hcy≥15 μmol/L的脑卒中复发率明显升高（Log-rank检验，P<0.05）。多因素Cox回归模型分析显示，高龄、Lg Hcy水平升高，以及房颤、糖尿病是脑卒中复发的独立危险因素。Lg Hcy分别与糖尿病、空腹血糖、年龄存在交互作用。 结论血浆Hcy水平升高既是高血压合并脑卒中患者卒中复发的独立危险因素，又通过与糖尿病、高龄、空腹血糖水平升高的交互作用显著增加脑卒中复发风险。     

The Chinese National Twin Registry: a ‘gold mine’ for scientific research

The Chinese National Twin Registry (CNTR) currently includes data from 61 566 twin pair from 11 provinces or cities in China. Of these, 31 705, 15 060 and 13 531 pairs are monozygotic, same‐sex dizygotic and opposite‐sex dizygotic pairs, respectively, determined by opposite sex or intrapair similarity. Since its establishment in 2001, the CNTR has provided an important resource for analysing genetic and environmental influences on chronic diseases especially cardiovascular diseases. Recently, the CNTR has focused on collecting biologic specimens from disease‐concordant or disease‐discordant twin pairs or from twin pairs reared apart. More than 8000 pairs of these twins have been registered, and blood samples have been collected from more than 1500 pairs. In this review, we summarize the main findings from univariate and multivariate genetic effects analyses, gene–environment interaction studies, omics studies exploring DNA methylation and metabolomic markers associated with phenotypes. There remains further scope for CNTR research and data mining. The plan for future development of the CNTR is described. The CNTR welcomes worldwide collaboration.     

RBMS3抑制宫颈癌细胞增殖和转移的作用机制研究

目的 研究RNA结合基序单链相互作用蛋白3(RBMS3)对宫颈癌细胞增殖和转移的影响,并探讨其发挥作用的可能机制.方法 采用GEPIA网站分析RBMS3在宫颈癌组织中的表达情况.qRT-PCR和Western blotting检测RBMS3 mRNA和蛋白在宫颈癌组织和细胞系中的表达水平.构建RBMS3过表达慢病毒,感染宫颈癌细胞系Caski,分为NC组和oe-RBMS3组,采用MTS检测各组细胞增殖能力,Transwell实验检测各组细胞转移能力,移植瘤实验检测各组细胞体内成瘤能力.Western blotting检测各组细胞中Wnt/β-catenin信号通路关键蛋白wnt3a、β-catenin及其下游靶蛋白cMYC、cyclinD1、MMP-2的表达.结果 GEPIA网站分析结果显示RBMS3 mRNA在宫颈癌组织中的表达显著低于在正常宫颈组织中的表达.qRT-PCR和Western blotting结果均显示RBMS3 mRNA和蛋白在宫颈癌组织中的表达显著低于其在癌旁组织中的表达,RBMS3 mRNA和蛋白在宫颈癌细胞系中的表达显著低于其在人鳞状上皮永生化细胞H8中的表达.与NC组相比,oe-RBMS3组Caski细胞增殖、转移和体内成瘤能力均降低,细胞中wnt3a、β-catenin、cMYC、cyclinD1、MMP-2蛋白表达均降低.结论 RBMS3可能通过调控Wnt/β-catenin信号通路抑制宫颈癌细胞增殖和转移能力.     

Structural insights into membrane remodeling by SNX1

The sorting nexin (SNX) family of proteins deform the membrane to generate transport carriers in endosomal pathways. Here, we elucidate how a prototypic member, SNX1, acts in this process. Performing cryoelectron microscopy, we find that SNX1 assembles into a protein lattice that consists of helical rows of SNX1 dimers wrapped around tubular membranes in a crosslinked fashion. We also visualize the details of this structure, which provides a molecular understanding of how various parts of SNX1 contribute to its ability to deform the membrane. Moreover, we have compared the SNX1 structure with a previously elucidated structure of an endosomal coat complex formed by retromer coupled to a SNX, which reveals how the molecular organization of the SNX in this coat complex is affected by retromer. The comparison also suggests insight into intermediary stages of assembly that results in the formation of the retromer-SNX coat complex on the membrane.

Sorting nexins (SNXs) exist as a large family of proteins defined by the presence of a PX (phox homology) domain (1, 2). Members of this family have been found to act as coat proteins in endosomal pathways that include recycling from endosomes to the plasma membrane and retrieval from endosomes to the Golgi complex (3, 4). Defects in these transport processes is associated with various neurologic disorders including Alzheimer’s disease, Parkinson’s disease, and Down’s syndrome (5, 6).Coat proteins assemble into complexes on the membrane to initiate intracellular transport pathways by coupling two main functions: bending the membrane to generate transport carriers and binding to cargoes for their sorting into these carriers (7). Retromer, a trimeric complex consisting of Vps26, Vps29, and Vps35, has been found to couple with different SNXs to form multiple endosomal coat complexes, in which select members of the SNX family act in membrane deformation while retromer acts in cargo recognition (8–17). Recently, a detailed molecular view of this functional cooperation has been achieved by elucidating the structure of a retromer-SNX complex on the membrane (18).Notably, it has been further discovered recently that an endosomal coat complex can be formed with only SNX members. SNX1/2 have been found to heterodimerize with SNX5/6 to form the endosomal SNX–BAR sorting complex for promoting exit 1 (ESCPE-1) complex, in which SNX1/2 are proposed to act in membrane deformation while SNX5/6 act in cargo recognition (19). As such, a key question has become whether SNX that acts in membrane deformation in this type of coat complex would be organized similarly on the membrane, as previously elucidated for SNX in the context of a retromer-SNX complex (18).One of the best characterized mechanisms of membrane deformation involves proteins that possess the BAR (Bin/Amphiphysin/Rvs) domain. This domain has been shown to undergo homodimerization to form a banana-shaped structure, which can impart membrane curvature through a scaffolding mechanism that involves electrostatic interactions between the positive charges lining the concave side of the curved BAR dimer and the negative charges that line the surface of the membrane bilayer. In some cases, the BAR domain can deform the membrane through a second mechanism, which involves the formation of an amphipathic helix that inserts into one leaflet of the membrane bilayer to generate bilayer asymmetry in driving membrane curvature (20, 21).Besides the PX domain, SNX1 also possesses a BAR domain. However, studies have found that its BAR domain is not sufficient in driving membrane deformation. Instead, the PX domain as well as the linker region between the BAR and PX domains are also needed (22, 23). As such, a key goal has been to achieve a better understanding of how the various parts of SNX1 contribute to its ability to deform the membrane.Structural studies, such as those involving crystallography and single-particle electron microscopy (EM), have been advancing a molecular understanding of coat proteins (24), including components of endosomal coats (17, 19, 22, 25–27). Notably, however, these approaches solve protein structures in solution, but the functional form of coat proteins involves their association with the membrane. In this study, we have pursued cryo-EM to reveal how SNX1 is organized on the membrane to explain its ability to deform the membrane. The result advances a molecular understanding of how an endosomal coat that contains only SNXs generates transport carriers. Moreover, by comparing our SNX1 structure to the previously solved retromer-SNX structure (18), we delineate the extent to which the molecular organization of SNX on the membrane is affected by the presence of retromer. This comparison also suggests insight into intermediary stages of coat assembly that form the retromer-SNX complex on the membrane.