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Pien Tze Huang Inhibits Migration and Invasion of Hepatocellular Carcinoma Cells by Repressing PDGFRB/YAP/CCN2 Axis Activity

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Chinese Journal of Integrative Medicine 2024年第2期30卷 115-124页

作者： LUO Zhi-yi TIAN Qi CHENG Niang-mei LIU Wen-han YANG Ye CHEN Wei ZHANG Xiang-zhi ZHENG Xiao-yuan CHEN Ming-sheng ZHUANG Qiu-yu ZHAO Bi-xing LIU Cong-sheng LIU Xiao-long LI Qin WANG Ying-chao The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province Mengchao Hepatobiliary Hospital of Fujian Medical UniversityFuzhou(350025)China Fujian Pien Tze Huang Enterprise Key Laboratory of Natural Medicine Research and Development Zhangzhou Pien Tze Huang Pharmaceutical Co.Ltd.ZhangzhouFujian Province(363099)China College of Biological Science and Engineering and Mengchao Med-X Center Fuzhou UniversityFuzhou(350116)China Department of Internal Medicine Mengchao Hepatobiliary Hospital of Fujian Medical UniversityFuzhou(350025)China

Objective:To investigate the effects of Pien Tze Huang(PZH) on the migration and invasion of HCC cells and underlying molecular ***:Cell counting kit-8(CCK-8) was applied to evaluate the cell viabilities of SMMC-7721,SK-Hep-1,C3A and HL-7702(6 × 10^(3)cells/well) co-incubated with different concentrations of PZH(0,0.2,0.4,0.6,0.8 mg/mL) for 24 ***,wound healing assay,CCK-8and Annexin V-FITC/PI staining were conducted to investigate the effects of PZH on the migration,invasion,proliferation and apoptosis of SK-Hep-1 and SMMC-7721 cells(650 μg/mL for SK-Hep-1 cells and 330 μg/mL for SMMC-7721 cells),*** vivo,lung metastasis mouse model constructed by tail vein injection of HCC cells was used for evaluating the anti-metastasis function of ***-Hep-1 cells(10^(6)cells/200 μL per mice) were injected into B-NDG mice via tail *** 8 mice were randomly divided into PZH and control groups,4 mice in each *** 2-d inoculation,mice in the PZH group were administered with PZH(250 mg/kg,daily) and mice in the control group received only vehicle(PBS) from the 2nd day after xenograft to day *** analysis based on RNA-seq was subsequently used for deciphering anti-tumor mechanism of *** real-time polymerase chain reaction(qRT-PCR) and Western blot were applied to verify RNA-seq *** reporter assay was performed to examine the transcriptional activity of yes-associated protein(YAP).Results:PZH treatment significantly inhibited the migration,invasion,proliferation and promoted the apoptosis of HCC cells in vitro and in vivo(Passociated with anti-metastasis function of *** study showed PZH significantly inhibited the expressions of platelet derived growth factor receptor beta(PDGFRB),YAP,connective tissue growth factor(CCN2),N-cadherin,vimentin and matrix metallopeptidase 2(MMP2,P<0.01).Meanwhile,the phosphorylatio

关键词： PienTzeHuang hepatocellular carcinoma RNA-seq Hippo yes-associated protein

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Inhibitory effects of dobutamine on human gastric adenocarcinoma

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World Journal of Gastroenterology 2014年第45期20卷 17092-17099页

作者： Hui-Xia Zheng Li-Na Wu Hong Xiao Qian Du Jian-Fang Liang Department of Pathology The First Hospital Shanxi Medical University

AIM: To explore the inhibitory effects of dobutamine on gastric adenocarcinoma cells.

关键词： Dobutamine Gastric adenocarcinoma cells yes-associated protein Hippo pathway Human gastric adenocarcinoma cell line SGC-7901 Therapy

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Adaptable hydrogel with reversible linkages for regenerative medicine:Dynamic mechanical microenvironment for cells

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Bioactive Materials 2021年第5期6卷 1375-1387页

作者： Zongrui Tong Lulu Jin Joaquim Miguel Oliveira Rui LReis Qi Zhong Zhengwei Mao Changyou Gao MOE Key Laboratory of Macromolecular Synthesis and Functionalization Department of Polymer Science and EngineeringZhejiang UniversityHangzhou310027China 3B’s Research Group I3Bs-Research Institute on BiomaterialsBiodegradables and BiomimeticsUniversity of MinhoHeadquarters of the European Institute of Excellence on Tissue Engineering and Regenerative MedicineAveParkZona Industrial da Gandra4805-017Barco GMRPortugal ICVS/3B’s-PT Government Associate Laboratory Braga/GuimarãesPortugal The Discoveries Centre for Regenerative and Precision Medicine Headquarters at University of MinhoAvepark4805-017BarcoGuimarãesPortugal Key Laboratory of Advanced Textile Materials&Manufacturing Technology Ministry of EducationNational Base for International Science and Technology Cooperation in Textiles and Consumer-Goods ChemistryZhejiang Sci-Tech University310018HangzhouChina

Hydrogels are three-dimensional platforms that serve as substitutes for native extracellular *** materials are starting to play important roles in regenerative medicine because of their similarities to native matrix in water content and *** would be very advantagoues for researchers to be able to regulate cell behavior and fate with specific hydrogels that have tunable mechanical properties as biophysical *** developments in dynamic chemistry have yielded designs of adaptable hydrogels that mimic dynamic nature of extracellular *** current review provides a comprehensive overview for adaptable hydrogel in regenerative medicine as ***,we outline strategies to design adaptable hydrogel network with reversible linkages according to previous findings in supramolecular chemistry and dynamic covalent ***,we describe the mechanism of dynamic mechanical microenvironment influence cell behaviors and fate,including how stress relaxation influences on cell behavior and how mechanosignals regulate matrix ***,we highlight techniques such as bioprinting which utilize adaptable hydrogel in regenerative *** conclude by discussing the limitations and challenges for adaptable hydrogel,and we present perspectives for future studies.

关键词： Adaptable hydrogel Dynamic mechanical microenvironment Supramolecular chemistry Dynamic covalent chemistry yes-associated protein

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Melatonin-based priming of stem cells to alleviate oxidative stress

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World Journal of Stem Cells 2024年第11期16卷 985-989页

作者： Khawaja Husnain Haider Department of Basic Sciences Sulaiman Al Rajhi UniversityAl Bukairiyah 51941AlQaseemSaudi Arabia

Stem cell expansion in vitro and transplantation in the cytokine-rich proinflammatory milieu in the injured tissue generate immense oxidative stress that interferes with the cells’survival,stemness,and *** cell priming has gained popularity to overcome these *** melatonin’s oxidative-scavenging properties,Gu et al have used periodontal ligament stem cells cultured under oxidative stress as an in vitro model to study the cytoprotective effects of *** letter to the editor delves into melatonin-induced stem cell priming and the underlying molecular mechanism,focusing on the intriguing role of yesassociated protein signaling in alleviating oxidative *** stress the importance of understanding the distinction between in vitro and in vivo oxidative stress conditions,a crucial aspect of stem cell research that invokes a sense of critical thinking in the *** study by Gu et al presents a novel approach to oxidative stress management,offering exciting possibilities for future research and applications.

关键词： Melatonin Oxidative stress Priming Stem cells yes-associated protein

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Molecular diagnosis of hepatocellular carcinoma:trends in biomarkers combination to enhance early cancer detection

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Hepatoma Research 2019年第3期5卷 20-33页

作者： Tiong Sun Chia Kwong Fai Wong John M.Luk Arbele Limited Hong Kong Science&Technology ParkShatinNew TerritoriesHong Kong

Despite of the advances in clinical imaging and applied research in proteomic biomarkers,liver cancer,especially hepatocellular carcinoma remains detected at the very late and advanced stages when curable treatments are unavailable and *** this regard,there are still huge unmet medical needs in developing and clinically validating those high-potential protein biomarkers preferably in liquid biopsy *** review provides a glimpse of emerging biomarkers together with detection tools and techniques which are potentially commercially available to the *** also discuss several diagnostic biomarkers having therapeutic potential for developing first-in-class medicines.

关键词： Hepatocellular carcinoma biomarkers targets α-fetoprotein cadherin-17 yes-associated protein AXL Trop2

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Resveratrol ameliorates lipopolysaccharide-induced anxiety-like behavior by attenuating YAP-mediated neuroinflammation and promoting hippocampal autophagy in mice

Resveratrol ameliorates lipopolysaccharide-induced anxiety-l...

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第七届全国呼吸系统重大疾病转化医学学术论坛

作者： Junming Fan Hui Guang Qiuyun Tian Yongyu Wang Xiaofang Fan Yongsheng Gong Institute of Hypoxia Medicine Wenzhou Medical University

AIM:Resveratrol（RSV） has been well documented to protect against inflammatory responses and exert neuroprotective ***-associated protein（YAP） is a major downstream effector of hippo signaling pathway that plays critical roles in ***,little is known about the role of RSV on YAP-mediated neuroinflammation associated with mental *** present study was sought to explore whether the anxiolytic effect of RSV was in relation to suppression of YAP pathway and identify the potential *** and RESULTS:We found that RSV significantly attenuated the anxietylike behavior deficits induced by lipopolysaccharide（LPS）,which was blocked by EX-527,a sirtuin 1（SIRT1） *** anxiolytic effect of RSV was accompanied by a marked decrease in the expression of YAP,IL-1β,and Iba-1, whereas RSV treatment can reverse the LPS-induced a decrease in autophagic proteins *** assay illustrated that SIRT1 functioned by counteracting with YAP *** study using XMU,a YAP activator,showed that activating YAP pathway potentiated LPS-induced anxiety-like behavior,while blockade of YAP activation with verteporfin,a YAP inhibitor,attenuated LPS-induced anxiety-like behavior by abrogating neuroinflammation and alleviating hippocampal autophagy caused by *** finally found that rapamycin-mediated promotion of autophagy attenuated LPS-induced anxiety-like behavior,whereas chloroquine,a lysosomal inhibitor,augmented autophagic flux and ameliorated LPS-induced anxietylike ***:Taken together,these results suggest that the amelioration of RSV in LPS-induced anxiety-like behavior is through attenuating YAP-mediated neuroinflammation and promoting hippocampal autophagy,and indicate the concept that inhibiting YAP pathway could be a potential therapeutic target for anxiety-like behavior during the process of neuroinflammation.

关键词： Resveratrol Lipopolysaccharide Anxiety-like behavior Autophagy yes-associated protein

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维替泊芬增强肺癌铂类耐药细胞A549/DDP顺铂敏感性的作用及机制研究

维替泊芬增强肺癌铂类耐药细胞A549/DDP顺铂敏感性的作用及机制研...

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作者：肖丹川北医学院

学位级别：硕士

研究背景:全球范围内,肺癌发病率及癌症相关死亡率均位居首位。肺癌主要以非小细胞肺癌(non-small cell lung cancer,NSCLC)为主,约占85%。尽管靶向药物已广泛应用于肺癌治疗,但对于无EGFR等突变的患者来说,铂类化疗药物如顺铂(Cisplati... 详细信息

研究背景:全球范围内,肺癌发病率及癌症相关死亡率均位居首位。肺癌主要以非小细胞肺癌(non-small cell lung cancer,NSCLC)为主,约占85%。尽管靶向药物已广泛应用于肺癌治疗,但对于无EGFR等突变的患者来说,铂类化疗药物如顺铂(Cisplatin,DDP),依然为一线化疗药物。但大量临床实践表明,长期铂类化疗会导致肺癌耐药,最终化疗失败、肿瘤进展。因此,探寻安全有效的顺铂化疗增敏剂并探明其作用机制,对克服NSCLC顺铂耐药并改善临床治疗效果极为重要。YAP(yes-associated protein)是控制组织器官大小的高保守Hippo信号通路的核心成员。研究表明,YAP在多种癌症中过表达,并与癌症干细胞自我更新、异常增殖、迁移侵袭以及预后不良相关,具有致癌基因作用。近来研究表明,YAP过度活化参与驱动铂类药物化疗耐药。抑制YAP可增强子宫颈癌、口腔鳞状细胞癌以及神经母细胞瘤对顺铂的敏感性。因此,靶向抑制YAP可能成为增强NSCLC顺铂化疗敏感性的潜在靶点。自噬(Autophagy)是将细胞内变性、衰老、受损的细胞器以及蛋白运送至溶酶体降解,并用以细胞循环再利用。正常情况下,自噬广泛存在于真核细胞中,对维持细胞稳态至关重要。在肿瘤发生发展过程中,自噬是一把双刃剑,可能促进肿瘤,也可能抑制肿瘤。近来研究表明,自噬可能是癌症化疗的保护机制,自噬抑制剂联合化疗药物抗肿瘤已成为新的研究热点。抑制自噬可增强肺癌、胃癌、结直肠癌以及卵巢癌对顺铂的敏感性。通过抑制自噬可能成为逆转NSCLC顺铂化疗耐药的潜在策略。维替泊芬(Verteporfin,VP)是由FDA批准的临床上用于治疗黄斑变性的第二代卟啉类光敏剂。最近研究表明,VP在抗肿瘤、逆转肿瘤耐药方面发挥着重要作用:VP直接抑制YAP表达,进而抑制卵巢癌增殖、膀胱癌侵袭转移、诱导胰腺癌细胞凋亡以及增强尿路上皮细胞癌细胞对顺铂的敏感性;VP通过抑制自噬增强胰腺导管腺癌对吉西他滨的敏感性;VP通过损伤溶酶体,阻断自噬流,增强肝细胞癌对索拉非尼的敏感性;此外,VP在结直肠癌中诱导p62形成寡聚体,而肿瘤细胞对高分子寡聚蛋白清除障碍,导致其具有肿瘤选择性蛋白毒性。那么,VP是否能增强肺癌铂类耐药细胞A549/DDP对顺铂的敏感性呢?其是否通过抑制YAP以及自噬实现逆转NSCLC顺铂耐药呢?因此,本研究旨在探讨VP对NSCLC A549/DDP细胞顺铂耐药性影响及其可能的分子机制,以期为维替泊芬作为非小细胞肺癌顺铂化疗增敏剂运用于临床提供实验室依据。研究目的:1.明确维替泊芬增强肺癌铂类耐药细胞A549/DDP顺铂敏感性的作用;2.探讨维替泊芬增强A549/DDP细胞顺铂化疗敏感性的潜在机制。研究方法:1.利用MTS、Western Blot、Real-time PCR检测维替泊芬联合顺铂处理A549/DDP细胞后,细胞增殖、凋亡相关蛋白Cleaved-Caspase3、Cleaved-PARP以及促凋亡基因PUMA、BIM的变化;2.利用细胞划痕、Transwell实验检测维替泊芬及顺铂处理A549/DDP细胞后,细胞迁移、侵袭能力的变化;*** Blot检测维替泊芬处理A549/DDP细胞后EMT相关蛋白E-cadherin、Vimentin、Snail 表达变化;*** Blot检测维替泊芬处理A549/DDP细胞后转运蛋白ABCG2、ABCG5表达变化。*** Blot检测A549、A549/DDP细胞中YAP蛋白的表达;维替泊芬处理A549/DDP细胞后YAP蛋白的表达变化;构建慢病毒感染稳定敲减YAP基因的A549/DDP细胞(A549/DDP-shYAP);分别应用Real-time PCR、Western Blot以及细胞免疫荧光检测shYAP对A549/DDP细胞中YAP的敲减效率;6.利用MTS、流式细胞仪检测A549/DDP-shYAP细胞顺铂处理后,细胞增殖、凋亡的变化;7.利用细胞划痕、Transwell实验检测A549/DDP-shYAP细胞顺铂处理后,细胞迁移、侵袭能力的变化;8.瞬转 pYAP 质粒在 A549 中过表达 YAP(A549-YAP),Western Blot验证过表达效果;*** Blot检测A549以及A549/DDP细胞EMT相关蛋白E-cadherin、Vimentin、Snail表达情况;分别检测A549细胞过表达YAP前后以及A549/DDP细胞敲减YAP前后EMT相关蛋白表达变化;*** Blot 检测 A549 以及 A549/DDP 细胞转运蛋白 ABCG2、ABCG5表达情况;分别检测A549细胞过表达YAP前后以及A549/DDP细胞敲减Y

关键词：非小细胞肺癌顺铂耐药维替泊芬 yes-associated protein 自噬

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Senolytics对人牙周膜干细胞干性维持、成骨分化和复制性衰老的作用及机制初探

Senolytics对人牙周膜干细胞干性维持、成骨分化和复制性衰老的作...

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作者：邱韵歌山东大学

学位级别：硕士

实验目的:本课题旨在探究Senolytics(槲皮素联合达沙替尼)对hPDLSCs干性维持、成骨分化和复制性衰老的调控作用,遴选槲皮素(Quercetin,Q)及达沙替尼(Dasatinib,D)联合用药的最佳作用浓度及配比,并且初步研讨Senolytics调控hPDLSCs生物... 详细信息

实验目的:本课题旨在探究Senolytics(槲皮素联合达沙替尼)对hPDLSCs干性维持、成骨分化和复制性衰老的调控作用,遴选槲皮素(Quercetin,Q)及达沙替尼(Dasatinib,D)联合用药的最佳作用浓度及配比,并且初步研讨Senolytics调控hPDLSCs生物学特性的可能作用机制。实验方法:选用组织块贴壁法获取hPDLSCs原代细胞;体外培养hPDLSCs并进行成骨、成脂诱导,利用碱性磷酸酶、茜素红、油红O染色,鉴定其多向分化潜能;结晶紫染色鉴定克隆及增殖能力;流式细胞术检测表面间充质干细胞标志物。细胞活性及细胞毒性检测技术(CCK8)分别检测Q、D对细胞的毒性作用,通过ALP(Alkaline phosphatase)活性检测、qRT-PCR检测成骨相关基因表达,筛选槲皮素最佳作用浓度,达沙替尼最佳作用浓度区间;ALP、茜素红染色鉴定Senolytics(D+Q)最佳配比浓度。通过Western Blot对成骨相关蛋白(COL]、ALP、RUNX2)的检测及ALP活性检测、ALP染色、茜素红染色验证Senolytics对于成骨分化的影响。通过Western Blot对干性维持相关蛋白(C-MYC、NANOG、OCT-4)的检测,验证Senolytics对于hPDLSCs干性维持的影响。选择生长状态良好的hPDLSCs在体外扩增到第20代(P20-hPDLSCs),建立自然衰老模型,取第5代hPDLSCs(P5-hPDLSCs)作为对照,进行衰老相关β-半乳糖苷酶(senescence associated β-galactosidase,SA-β-Gal)染色、qRT-PCR 检测衰老相关基因 p16、p21的表达情况,验证Senolytics对于hPDLSCs自然衰老的影响。通过Western Blot检测Senolytics对于YAP/TAZ的影响;通过慢病毒转染建立特异性敲低TAZ模型,ALP活性检测、ALP染色、茜素红染色、Western Blot检测成骨相关蛋白表达验证TAZ在Senolytics促进hPDLSCs成骨中的作用。使用YAP抑制剂维替泊芬(Verteporfin)抑制hPDLSCs中YAP的表达,Western Blot检测干性相关蛋白表达,验证YAP在Senolytics促进hPDLSCs干性维持中的作用。实验结果:成功分离提取符合后续实验要求的hPDLSCs,并在体外有效扩增。一定浓度区间内的槲皮素能有效促进hPDLSCs的成骨分化,最佳单药浓度为2.5 uM;浓度≤1 nM达沙替尼对hPDLSCs成骨有促进作用,浓度>1 nM对成骨有抑制作用,最佳单药浓度为1 nM。Senolytics联合用药与单一用药相比,增强了槲皮素的促成骨作用,并改变了高浓度达沙替尼抑制成骨的现象,最终选择配比浓度D(1 nM)+Q(2.5 uM)以及D(10 nM)+Q(2.5 uM)用于后续实验。Senolytics促进相关成骨蛋白的表达,促进ALP活性、ALP染色及茜素红矿化结节的形成。Senolytics促进相关干性维持蛋白的表达。Senolytics通过清除衰老细胞,有效下调衰老相关基因表达及β-半乳糖苷酶染色阳性细胞比例。Senolytics上调了 YAP/TAZ的蛋白表达;敲低TAZ后,部分逆转了 Senolytics对于hPDLSCs的成骨促进作用;抑制YAP表达后部分逆转了 Senolytics对于hPDLSCs的干性维持的正向调控作用。实验结论:Senolytics(D+Q)对于hPDLSCs的成骨分化及干性维持有正向调控作用,并且可以一定程度抵抗hPDLSCs的复制性衰老;Hippo-YAP/TAZ参与调控Senolytics对于hPDLSCs生物学特性的影响。

关键词：人牙周膜干细胞 Senolytics 槲皮素达沙替尼成骨分化干性维持衰老 yes-associated protein transcriptional coactivator with PDZ binding motif

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