shRNA沉默干扰HMGA2基因对胃癌细胞株MKN-45的增殖与凋亡的影响

目的:观察小分子RNA(shRNA)沉默后HMG-A2基因在胃癌细胞株MKN-45的表达,并探讨HMGA2基因对胃癌细胞的增殖与凋亡的影响.方法:构建针对人HMGA2基因的shRNA真核表达载体,瞬时转染人胃癌细胞株MKN-45.用细胞免疫组化的方法观察转染72h后HMGA2的蛋白表达水平,以MTT比色法、流式细胞术检测转染后MKN-45细胞的生长增殖、凋亡的情况.结果:转染HMGA2-shRNA组的蛋白表达强度(171.34±19.61)明显弱于scrambled组(143.48±19.04)和空白对照组(141.79±18.09,P<0.05),较之scrambled组(5.66%±0.63%)和空白对照组,HMGA2-shRNA组(39.32%±2.37%)能明显抑制细胞的增殖(P<0.05),HMGA2-shRNA组的凋亡率(39.67%±2.35%)与scrambled组(4.29%±1.33%)和空白对照组(5.05%±1.84%)相比明显增加(P<0.05).结论:靶向HMGA2基因的shRNA可以有效抑制人胃癌MKN-45细胞的生长,并促进细胞的凋亡,HMGA2可能是胃癌治疗的一个潜在靶点.     

RNAi沉默Sp3基因对肝癌HepG2细胞增殖的影响

目的:采用基因沉默和慢病毒转染技术,沉默人肝癌细胞HepG2中特异蛋白3(specificity protein 3,Sp3)的表达,观察沉默Sp3基因后的肝细胞的增殖能力变化.方法:采用Sp3-siRNA慢病毒转染人肝癌细胞HepG2,通过免疫印迹法和PCR技术检测Sp3的表达以验证转染效果,通过MTT检测细胞生长曲线和流式细胞技术测定细胞周期.结果:Western blot实验表明,实验组的Sp3表达量明显少于空白组和阴性对照组(0.37±0.08 vs 0.83±0.17,0.66±0.13,F=8.442,均P<0.05).RT-PCR也得到相同的结果(0.47±0.05 vs 0.74±0.08,0.70±0.16,F=7.322,均P<0.05).MTT实验结果显示,与空白对照和阴性对照组相比,实验组细胞在48、72和96h时的增殖明显受到抑制(0.28±0.18 vs 0.34±0.19,0.35±0.07,F=3.888;0.57±0.11 vs 0.84±0.05,0.74±0.08,F=12.721;0.72±18.1 vs 0.98±0.05,0.93±0.9,F=6.342,均P<0.05).流式细胞术结果显示实验组细胞主要分布在G1期.结论:RNAi沉默Sp3基因导致人肝癌HepG2细胞体外增殖能力下降.     

Perspective: physiological role(s) of the vascular myogenic response

The vascular myogenic response is an inherent property of VSM in the walls of small arteries and arterioles, allowing these principal resistance segments of the microcirculation to respond to changes in transmural pressure. Elevated intraluminal pressure leads to myogenic constriction, whereas reduced pressure leads to myogenic dilation. This review focuses on the physiological significance of the myogenic response in microvascular networks. First, historical concepts related to the detection of stretch by the vessel wall are reviewed, including the wall tension hypothesis, and the implications of the proposal that the arteriolar network responds to Pp changes as a system of series-coupled myogenic effectors. Next, the role of the myogenic response in the local regulation of blood flow and/or Pc is examined. Finally, the interaction of myogenic constriction and dilation with other local control mechanisms, including metabolic, neural and shear-dependent mechanisms, is discussed. Throughout the review, an attempt is made to integrate historical and current literature with an emphasis on the physiological role, rather than the underlying signaling mechanisms, of this important component of vascular control.     

环状非编码RNA在眼部疾病中表达研究进展

环状非编码RNA（circRNA）是指一类不具有5’末端帽子和3’末端poly（A）尾巴的一类非编码RNA,其通过共价键形成环形结构,它们可以通过几种不同的机制调节蛋白质的编码。越来越多的研究发现,circRNA在细胞增殖、分化、凋亡、血管生成、免疫调节等多种细胞功能中发挥一定的作用,从而导致多种疾病（如恶性肿瘤、炎症性疾病、神经系统疾病等）的发生。本文综述了circRNA在眼科疾病中的潜在作用和可能机制,以期能够为相关眼科疾病的早期诊断和不同阶段的治疗提供新的思路。     

Single-cell RNA sequencing in vision research: Insights into human retinal health and disease

Gene expression provides valuable insight into cell function. As such, vision researchers have frequently employed gene expression studies to better understand retinal physiology and disease. With the advent of single-cell RNA sequencing, expression experiments provide an unparalleled resolution of information. Instead of studying aggregated gene expression across all cells in a heterogenous tissue, single-cell technology maps RNA to an individual cell, which facilitates grouping of retinal and choroidal cell types for further study. Single-cell RNA sequencing has been quickly adopted by both basic and translational vision researchers, and single-cell level gene expression has been studied in the visual systems of animal models, retinal organoids, and primary human retina, RPE, and choroid. These experiments have generated detailed atlases of gene expression and identified new retinal cell types. Likewise, single-cell RNA sequencing investigations have characterized how gene expression changes in the setting of many retinal diseases, including how choroidal endothelial cells are altered in age-related macular degeneration. In addition, this technology has allowed vision researchers to discover drivers of retinal development and model rare retinal diseases with induced pluripotent stem cells. In this review, we will overview the growing number of single-cell RNA sequencing studies in the field of vision research. We will summarize experimental considerations for designing single-cell RNA sequencing experiments and highlight important advancements in retinal, RPE, choroidal, and retinal organoid biology driven by this technology. Finally, we generalize these findings to genes involved in retinal degeneration and outline the future of single-cell expression experiments in studying retinal disease.     

靶向干扰细胞周期检验点激酶2增强5-氟尿嘧啶对鼻咽癌细胞SUNE-1的毒性作用研究

目的探讨靶向抑制细胞周期检验点激酶2（Chk2）对5-氟尿嘧啶（5-FU）抗鼻咽癌鳞状上皮细胞SUNE-1的增敏作用。方法根据Chk2基因设计siRNA干扰序列,在mRNA和蛋白水平检测其干扰效果;通过细胞生长曲线观察Chk2siRNA对SUNE-1细胞增殖能力的影响;通过细胞毒性试验检测Chk2siRNA是否影响SUNE-1细胞对代谢类抗肿瘤药5-FU的敏感性;采用AnnexinV／PI双染方法检测细胞凋亡的变化情况。结果所采用的siRNA在mRNA和蛋白水平均显著降低SUNE-1细胞中Chk2表达,验证了该siRNA的干扰效果;Chk2siRNA对SUNE-1细胞增殖能力无显著影响,但能显著增强5-Fu的细胞毒作用（P〈0．05）;细胞凋亡检测结果显示,Chk2siRNA显著增加5-FU引起的SUNE-1细胞凋亡水平。结论尽管Chk2对SUNE-1细胞生存和增殖不发挥关键作用,但在5-Fu引起的细胞损伤中对细胞起保护作用,该作用与其抑制5-FU引起的细胞凋亡有关,因此Chk2可成为在鼻咽癌治疗中增强5-FU的疗效的靶点。     

角膜新生血管中细胞与分子的研究进展

多种眼部损伤可诱导角膜新生血管形成,促进疾病发展,造成角膜水肿、视力受损,甚至失明,因此抑制角膜新生血管有助于延缓疾病进程并降低角膜损伤,具有十分重要的临床意义。本文将对参与角膜新生血管形成的细胞及分子作最新的系统论述,并分析可能的抑制靶点,以期为科研及临床提供参考。     

微小RNA和衰老

微小RNA（miRNA）是一类非编码的单链小分子RNA,不仅是机体有目的编码RNA的重要调控分子,并且参与生长、发育、衰老的调控。本文主要描述了miRNA参与的有关衰老调控的一些可能通路和相关机制,如调控端粒酶的表达、p53信号通路、Rb通路;另外,本文还综述了通过调控miRNA合成的酶而影响衰老的一些特殊途径,以阐明当前miRNA与衰老关系的研究成果。