Hippo信号通路在后发性白内障中的研究进展

后发性白内障又称后囊膜混浊(posterior capsular opacification,PCO)是白内障摘除术后最为常见的并发症,严重影响患者的生活质量。目前尚无防止其发生的有效措施,临床上多用手术或Nd：YAG激光治疗,现在亟需新的治疗方案。Hippo信号通路参与了哺乳动物多种细胞和器官的稳态调控,最近研究表明Hippo信号通路可调控晶状体上皮细胞(LECs)增殖、凋亡、分化等行为,Hippo信号通路可能为后发性白内障治疗提供新的靶点,本文论述了Hippo信号通路的组成、调节机制及其在后发性白内障中与其他信号通路的相互关系及相关药物治疗,以期为后发性白内障防治提供更广阔的思路。     

siRNA沉默YAP1对宫颈癌Hela细胞增殖和侵袭的影响

目的:用小干扰RNA(siRNA)沉默宫颈癌Hela细胞中YAP1基因的表达,观察YAP1表达降低对细胞增殖和侵袭能力的影响,初步探讨YAP1在宫颈癌中的作用。方法:qRT-PCR和Western blot方法检测宫颈癌Hela细胞及正常宫颈上皮细胞H8中YAP1的表达,siRNA YAP1转染Hela细胞沉默YAP1表达,并以转染control siRNA的细胞作为对照,转染48h后,分别用qRT-PCR和Western blot方法检测沉默效果。用MTT法检测沉默后细胞增殖情况。Transwell 检测细胞侵袭能力。结果:YAP1在宫颈癌细胞中高表达。siRNA能够有效沉默Hela细胞中YAP1的mRNA 及蛋白的表达。沉默YAP1表达能够明显抑制Hela细胞的增殖及侵袭能力。结论:YAP1蛋白在宫颈癌细胞中高表达并具有促进细胞增殖及侵袭的能力。     

RNA干扰YAP基因对人膀胱癌T24细胞株增殖和迁移能力的影响

背景与目的：膀胱尿路上皮癌是泌尿统系最常见的肿瘤,术后易复发及转移,Yes相关蛋白(Yes associated protein,YAP)基因与膀胱癌关系密切,本研究探讨通过RNA干扰技术沉默膀胱癌T24细胞中YAP基因的表达,观察YAP基因沉默后对人膀胱癌T24细胞增殖、迁移能力的影响。方法：用阳离子脂质体转染试剂LipofectamineTM2000将靶向沉默YAP基因的小干扰RNA(small interfering RNA,siRNA)序列转染至膀胱癌T24细胞株中,采用实时定量逆转录聚合酶链反应(quantitative real time-polymerase chain reaction,qRT-PCR)、蛋白质印迹法(Western blot)检测转染后T24细胞中YAP基因及蛋白的表达水平,细胞增殖活性检测试剂盒(cell counting kit-8,CCK-8)、Transwell迁移试验以及划痕实验观察siRNA在体外对人膀胱癌T24细胞增殖、迁移能力的影响。结果：转染siRNA后,YAP RNA和蛋白表达量同空白对照以及阴性对照组相比被显著抑制(RNA：F=93.91,P<0.000 1;蛋白：F=4.62,P<0.05),CCK-8增殖活性实验结果显示,RNAi干扰YAP表达可以显著抑制膀胱癌T24细胞的增殖活性(12 h：F=6.00,P=0.037;24 h：F=41.72,P=0.000 3;36 h：F=462.8,P<0.000 1;48 h：F=236.6,P<0.000 1;72 h：F=140.5,P<0.000 1),通过Transwell实验和划痕试验发现,RNAi干扰YAP表达显著抑制膀胱癌T24细胞的迁移能力(Transwell：F=43.55,P<0.05;划痕：F=43.55,P<0.05)。结论：YAP基因是膀胱肿瘤增殖及迁移的重要调控因子。     

Interactions between Src family protein tyrosine kinases and PSD-95

Five members of the Src family of non-receptor protein tyrosine kinases--Lck, Lyn, Fyn, Src, and Yes--are known to be expressed in the central nervous system. Src and Fyn have been shown to play important roles in synaptic transmission and plasticity at excitatory synapses. Here we investigate the subcellular distribution and potential binding partners of Src family protein tyrosine kinases in brain, focusing on the lesser studied kinases Lck, Lyn, and Yes. We find that Lck, Lyn, and Yes are localized to the postsynaptic density (PSD), the primary structural component of excitatory synapses. Lyn and Yes, as well as Src, but not Lck physically associate with the prominent PSD scaffolding protein PSD-95 in co-immunoprecipitation experiments. Further, we demonstrate that PSD-95 GST fusion proteins bind directly to purified recombinant Lyn, Src, and Yes in vitro. In addition, we show that PSD-95 is unique among PSD-95 family members in that the other members, PSD-93, SAP97, and SAP102, do not physically associate with Lyn, Src, or Yes. Together our results suggest that PSD-95 may be important for localizing and/or regulating multiple Src protein tyrosine kinases at the NMDA receptor multiprotein complex.     

Association between YAP expression in neoplastic and non‐neoplastic breast tissue with arsenic urinary levels

The Hippo pathway regulates cell proliferation and apoptosis and it has been noted that loss of critical components of this pathway can lead to uncontrolled cell growth. Yes‐associated protein (YAP) is an important component of this Hippo pathway because YAP is the nuclear effector of the Hippo tumor suppressor pathway and it is crucial for the response to oxidative stress induced by cellular process and by different xenobiotics, including arsenic. It has been proposed that YAP dysregulation can contribute to a malignant cellular phenotype acting as both a tumor suppressor and an oncogene. The aim of the study was to assess and compare the expression of YAP in neoplastic and non‐neoplastic breast tissue of women chronically exposed to arsenic through drinking water. YAP expression was assessed by immunohistochemistry in 120 breast biopsies from women with breast cancer and from women with other non‐neoplastic breast pathologies. Arsenic concentration was quantified in urine. The results disclosed a significant lower percentage of cytoplasm YAP expression in cases and that YAP high‐intensity staining in the cytoplasm but not in the nucleus decreases the risk for breast cancer. In conclusion, our overall data suggest that YAP may act as a tumor suppressor protein because their reduced expression in cases, which can induce an environment favorable for inhibition of apoptosis and promoting cellular proliferation by increasing genetic instability of cells, which might contribute to the pathogenesis of cancer. Copyright © 2017 John Wiley & Sons, Ltd.     

沉默Yes相关蛋白1对人非小细胞肺癌细胞增殖和侵袭的影响

目的 研究Yes相关蛋白1(YAP1)在非小细胞肺癌细胞增殖、细胞周期、迁移、侵袭中的作用.方法 通过慢病毒介导的小干扰RNA靶向抑制人非小细胞肺癌细胞株A549中YAP1基因的表达,采用CCK-8法、流式细胞术分别检测沉默YAP1对A549细胞增殖及凋亡周期的影响,Transwell实验观察沉默YAP1对A549细胞迁移、侵袭能力的影响.结果 沉默YAP1后,A549细胞YAP1 mRNA和蛋白表达均下调(P＜0.01),CCK-8实验结果显示沉默YAP1抑制A549的增殖、增加G0/G期的细胞比例(P＜0.01),Transwell实验结果示沉默YAP1抑制A549细胞的迁移、侵袭(P＜0.01).结论 沉默YAP1基因对非小细胞肺癌细胞的恶性生物学特征具有抑制作用,YAP1可能成为肺癌治疗的潜在靶标.