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Cytosolic phospholipase A2α modulates cell-matrix adhesion via the FAK/paxillin pathway in hepatocellular carcinoma

Cancer Biology & Medicine 2019年第2期16卷 377-390页

作者： Piao Guo Yuchao He Lu Chen Lisha Qi Dongming Liu Ziye Chen Manyu Xiao Liwei Chen Yi Luo Ning Zhang Hua Guo Department of Tumor Cell Biology Department of Hepatobiliary Cancer Department of Pathology Tianjin Medical University Cancer Institute and Hospital National Clinical Research Center for Cancer/ Key Laboratory of Cancer Prevention and Therapy Tianjin Tianjin's Clinical Research Center for Cancer Tianjin 300060 China

Objective: To explore the effect of cytosolic phospholipase A2α(cPLA2α) on hepatocellular carcinoma(HCC) cell adhesion and the underlying ***: Cell adhesion, detachment, and hanging-drop assays were utilized to examine the effect of cPLA2α on the cell-matrix and cell-cell adhesion. Downstream substrates and effectors of cPLA2α were screened via a phospho-antibody *** signaling pathways were identified by the functional annotation tool DAVID. Candidate proteins were verified using Western blot and colocalization was investigated via immunofluorescence. Western blot and immunohistochemistry were used to detect protein expression in HCC tissues. Prognosis evaluation was conducted using Kaplan-Meier and Cox-proportional hazards regression ***: Our findings showed that cPLA2α knockdown decreases cell-matrix adhesion but increases cell-cell adhesion in HepG2 cells. Microarray analysis revealed that phosphorylation of multiple proteins at specific sites were regulated by cPLA2α. These phosphorylated proteins were involved in various biological processes. In addition, our results indicated that the focal adhesion pathway was highly enriched in the cPLA2α-relevant signaling pathway. Furthermore, cPLA2α was found to elevate phosphorylation levels of FAK and paxillin, two crucial components of focal adhesion. Moreover, localization of p-FAK to focal adhesions in the plasma membrane was significantly reduced with the downregulation of cPLA2α. Clinically, cPLA2α expression was positively correlated with p-FAK levels. Additionally, high expression of both cPLA2α and p-FAK predicted the worst prognoses for HCC ***: Our study indicated that cPLA2α may promote cell-matrix adhesion via the FAK/paxillin pathway, which partly explains the malignant cPLA2α phenotype seen in HCC.

关键词： Hepatocellular carcinoma cytosolic phospholipase A2α cell-matrix adhesion FAK paxillin

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paxillin serine 178 phosphorylation in control of cell migration and metastasis formation through regulation of EGFR expression in breast cancer

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Journal of Cancer Metastasis and Treatment 2019年第6期4卷 71-86页

作者： Saertje Verkoeijen Ya-Feng Ma Wies van Roosmalen Reshma Lalai Martine H.A.M.van Miltenburg Marjo de Graauw Bob van de Water Sylvia E.Le Dévédec Institute for Life Sciences&Chemistry Hogeschool Utrecht HUUtrecht 3501 AAthe Netherlands Division of Drug Discovery and Safety Leiden Academic Centre for Drug ResearchLeiden UniversityLeiden 2300 RAthe Netherlands

Aim:paxillin is a well-known multidomain scaffold protein that is involved in the regulation of cell-matrix adhesion dynamics,a process required for the tumor cell migration and *** of the serine residue 178 requires c-Jun NH2-terminal kinase(JNK)activation,which occurs downstream of epidermal growth factor receptor(EGFR)-mediated signaling and drives cell *** this study,we investigated the significance of paxillin Ser178 phosphorylation in breast cancer ***:We employed the rat mammary carcinoma MTLn3 cell line with which we established stabile variants of both wild type and mutant GFP-paxillin *** those,we next performed several in vitro assays including cell proliferation,migration and focal adhesion ***,we monitored the metastatic spread of both cell line variants in an othrotopic mouse model for breast ***:Here we show that expression of the phospho-defective mutant paxillinS178A in the metastatic mammary adenocarcinoma MTLn3 cell-line significantly decreased EGF-induced cell migration,which was correlated with impaired focal adhesion ***,paxillinS178A attenuated lung metastasis formation in an orthotopic in vivo mammary gland tumor/metastasis model,demonstrating the importance of JNK-mediated paxillin phosphorylation in breast cancer *** of paxillinS178A caused a decrease in EGFR expression, ;while re-expression of EGFR in MTLn3-paxillinS178A cells fully restored EGF-driven cell motility and focal adhesion ***,re-expression of EGFR in MTLn3-paxillinS178A rescued spontaneous metastasis from breast to ***:Overall our data show an important role for JNK-mediated paxillin Ser178 phosphorylation in the regulation of EGFR expression and thereby,in EGF-driven cell migration and metastasis formation.

关键词： paxillin c-Jun NH2-terminal kinase focal adhesion epidermal growth factor receptor cell migration metastasis breast cancer

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paxillin在高、低转移潜能乳腺癌中的表达及意义

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四川大学学报（医学版） 2010年第1期41卷 91-94页

作者：蔡辉张涛汤为学李少林重庆医科大学基础医学院放射医学教研室重庆400016 重庆医科大学附属第一医院肿瘤科重庆400016 重庆医科大学基础医学院病理生理学教研室重庆400016

目的研究paxillin在人高、低转移潜能乳腺癌细胞亚系中的表达以及对肿瘤转移、黏附的影响。方法以乳腺癌细胞系MDA-MB-435S为研究对象,利用细胞穿透人工基底膜能力的差异筛选出高、低转移潜能的乳腺癌细胞系,利用免疫组化、免疫印迹(Wes... 详细信息

目的研究paxillin在人高、低转移潜能乳腺癌细胞亚系中的表达以及对肿瘤转移、黏附的影响。方法以乳腺癌细胞系MDA-MB-435S为研究对象,利用细胞穿透人工基底膜能力的差异筛选出高、低转移潜能的乳腺癌细胞系,利用免疫组化、免疫印迹(Western blot)、逆转录聚合酶链反应(RT-PCR)检测paxillin在高、低转移潜能乳腺癌细胞中的表达。采用四甲基偶氮唑蓝(MTT)法检测细胞黏附率。结果免疫组化、Western blot、RT-PCR检测显示在高转移潜能乳腺癌细胞中paxillin在基因和蛋白水平的表达均高于低转移潜能乳腺癌细胞,差异具有统计学意义(P均paxillin在人乳腺癌转移和黏附过程中发挥重要作用,可能成为抑制肿瘤增殖和转移的关键性分子。

关键词：乳腺癌 paxillin 肿瘤转移

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Expression and localization of paxillin in rat pancreas during development

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World Journal of Gastroenterology 2011年第40期17卷 4479-4487页

作者： Jing Guo Li-Jie Liu Li Yuan, Ning Wang Wei De Department of Biochemistry and Molecular Biology Nanjing Medical University Nanjing 210029 Jiangsu Province China Department of Physiology Southeast University Nanjing 210009 Jiangsu Province China

AIM:To investigate the expression and localization of paxillin in rat pancreas during development. METHODS:Pancreata from Sprague Dawley rat fetuses, embryos, young animals, and adult animals were used in this study. Expression levels of paxillin in pancreata of different development stages were detected by reverse transcription polymerase chain reaction and Western blotting. To identify the cell location of paxillin in the developing rat pancreas, immunohistochemistry and double-immunofluorescent staining were performed using antibodies for specific cell markers and paxillin, respectively. RESULTS:The highest paxillin mRNA level was detected at E15.5 (embryo day 15.5) following a decrease in the later developmental periods (P paxillin protein expression through the transition from E15.5 to adult was detected. The paxillin positive staining was mainly localized in rat islets of Langerhans at each stage tested during pancreas development. CONCLUSION:The dynamic expression of paxillin in rat pancreas from different stages indicates that paxillin might be involved in some aspects of pancreatic development.

关键词： Pancreas development Islet remodeling paxillin Cell adhesion Migration

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Role of paxillin in colorectal carcinoma and its relationship to clinicopathological features

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Chinese Medical Journal 2014年第3期127卷 423-429页

作者： Yin Hongfang Zhang Quanwen Wang Xin Li Ting Wan Yuanlian Liu Yucun Zhu Jing Department of Pathology Peking University First Hospital Beijing 100034 China Department of General Surgery Peking University First Hospital Beijing 100034 China Surgical Laboratory Peking University First Hospital Beijing 100034 China

Background Colorectal carcinoma is one of the most common malignant *** advances in therapy,mortality is still very *** aim of this study was to evaluate the expression of paxillin in the human colon adenocarcinoma cell line SW480 and its role in cell cycle and *** also investigated the expression of paxillin in colorectal carcinoma tissues and its relationship to clinicopathological features and *** paxillin short hairpin RNA （shRNA） was constructed and transfected into the colon adenocarcinoma cell line *** influence of paxillin shRNA on the cell cycle and cell apoptosis was analyzed by flow *** staining was used to assess the expression of paxillin and its association with the expression of carcinoembryonic antigen （CEA）,carbohydrate antigen （CA） 19-9,p53 and Bcl-2 in 102 patients with primary colorectal *** blotting was also used to investigate the expression of *** records were reviewed and a clinicopathological analysis was *** In vitro,the percentage of cells in S phase was （45.23±1.05）％,（43.53±1.23）％,and （36.13±0.57）％ in the blank control group,negative control group,and paxillin shRNA group *** was significantly decreased in the paxillin shRNA group （P=0.000）.The early apoptosis index of the paxillin shRNA group （17.2±1.18％） was significantly increased compared to the control shRNA group （（13.17±1.15）％,P=0.013）.paxillin was positive in 71 （69.6％） patients,and it was found to be overexpressed in tumor tissues compared with normal adjacent *** positive rate was higher in patients who are less than 50-years old （100.0％ vs.65.6％,P=0.016）.paxillin expression was associated with a high histologic grade of carcinoma （81.4％ vs.61.0％,P=0.031）,a high rate of regional lymph node metastasis （22.5％ vs.13.0％,P=0.031）,mesenteric artery lymph node metastasis （100.0％ vs.64.8％,P=0.008）,distant metastasis （94.1％ vs.64.7％,P=-0.016） and a high

关键词： cell cycle apoptosis colorectal neoplasms paxillin clinicopathology

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Clinicopathological significance of expression of paxillin, syndecan-1 and EMMPRIN in hepatocellular carcinoma

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World Journal of Gastroenterology 2005年第10期11卷 1445-1451页

作者： Hai-GangLi De-RongXie Xi-MingShen Hong-HaoLi HongZeng Yun-JieZeng DepartmentofPathology TheSecondAffiliatedHospitaltoSunYat-SenUniversityGuangzhou510120GuangdongProvinceChina DepartmentofOncology TheSecondAftiliatedHospitaltoSunYat-SenUniversityGuangzhou510120GuangdongProvinceChina DepartmentofSurgery TheSecondAffiliatedHospitaltoSunYat-SenUniversityGuangzhou510120GuangdongProvinceChina

AIM: To evaluate the relationship of expression of paxillin,syndecan-1 and EMMPRIN proteins with clinicopathological features in hepatocellular carcinoma (HCC).METHODS: Fifty-one patients who underwent HCC resection were recruited in the study. paxillin, syndecan1 and EMMPRIN proteins in HCC tissues were detected with immunohistochemical ***: Of 51 cases of HCC, 23 (45%) exhibited paxillin protein positive expression. Of 42 cases of adjacent nontumor liver tissues, 24 (57%) exhibited positive *** paxillin protein expression was associated with low differentiation (r= 0.406, P= 0.004), with the presence of portal vein thrombosis (r = 0.325, P = 0.021), with extra-hepatic metastasis (r = 0.346, P = 0.014). Of 51cases of HCC, 28 (55%) exhibited syndecan-1 protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 23 (55%) exhibited positive *** snydecan-1 protein expression was associated with well differentiation (r = 0.491, P = 0.001), with no extra-hepatic metastasis (r = 0.346, P = 0.014). Of 51cases of HCC, 28 (55%) exhibited EMMPRIN protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 21 (50%) exhibited positive *** of EMMPRIN protein was not associated with serum AFP level, HBsAg status, presence of microsatellite nodule, tumor size, presence of cirrhosis and necrosis,differentiation, presence of portal vein thrombosis, extrahepatic metastasis, disease-free survival and overall survival (P＞0.05). Expression of paxillin protein was correlated conversely with the expression of syndecan-1protein in HCC (r = -0.366, P = 0.010).CONCLUSION: Expression of paxillin and syndecan-1proteins in HCC may affect its invasive and metastatic ability of the tumor. There may be a converse correlation between the expression of paxillin and syndecan-1 protein in HCC. Expression of EMMPRIN protein may be detected in HCC, but it may play little role in the invasion and metastasis

关键词： Hepatocellular carcinoma paxillin Syndecan-1 EMMPRIN Immunohistochemistry

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paxillin对骨髓间充质干细胞向类髓核细胞分化的影响

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中国脊柱脊髓杂志 2016年第5期26卷 453-458页

作者：刘汝银岳宗进彭晓艳王新立冯仲锴河南省中医院脊柱科郑州市450002 河南省中医院护理部郑州市450002

目的 :探讨paxillin对骨髓间充质干细胞(BMSCs)向类髓核细胞分化的影响。方法 :取3周龄健康SD大鼠骨髓,采用全骨髓贴壁法培养BMSCs,传至第3代,用流式细胞仪对其表面标志分子进行鉴定。利用pc DNA3.1(-)/Myc-His B质粒构建真核表达载体pc... 详细信息

目的 :探讨paxillin对骨髓间充质干细胞(BMSCs)向类髓核细胞分化的影响。方法 :取3周龄健康SD大鼠骨髓,采用全骨髓贴壁法培养BMSCs,传至第3代,用流式细胞仪对其表面标志分子进行鉴定。利用pc DNA3.1(-)/Myc-His B质粒构建真核表达载体pc DNA3.1-paxillin。取第3代BMSCs分为3组:实验组,转染pc DNA3.1-paxillin;阴性对照组,转染pc DNA3.1(-)/Myc-His B;空白对照组,不转染质粒。分别培养36h后,实验组和阴性对照经G418筛选,取3组BMSCs采用实时荧光定量PCR方法(q RT-PCR)检测各组paxillin、蛋白多糖(Aggrecan)、Ⅱ型胶原α1(collagen typeⅡα1,COL2α1)、性别决定区Y框蛋白9(sex determining region Y box protein 9,SOX9)、角蛋白19(keratin19,KRT19)、配对盒1(paired box 1,PAX1)和叉头蛋白(forkhead box F1,FOXF1)的m RNA表达量;免疫印迹法(Western blot)检测paxillin、KRT19、PAX1和FOXF1蛋白表达水平。结果:BMSCs原代培养至第3代的细胞形态大体一致,呈长梭形,细胞连接紧密且均匀分布,CD44、CD90呈现高表达(95.6%和96.8%),CD45、CD11则呈现低表达(1.82%和2.16%)。实验组中paxillin的m RNA和蛋白表达量较阴性对照组和空白对照组升高,差异具有显著性(Ppaxillin可以促进BMSCs向类髓核细胞分化,为椎间盘退变性疾病的细胞移植治疗提供基础。

关键词：骨髓间充质干细胞 paxillin 类髓核细胞真核表达载体

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paxillin和p130Cas蛋白在肝癌中的表达及意义

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中华外科杂志 2006年第23期44卷 1648-1649页

作者：潘胜利陈孝平张万广李高鹏华中科技大学同济医学院附属同济医院肝脏外科中心

肝癌复发和转移是一个复杂的生物学过程,涉及多个基因和多种分子变化.研究表明,细胞的黏附动力学和形态改变在肿瘤的侵袭与转移中起重要作用.paxillin(桩蛋白)和p130Cas是两个黏着斑相关蛋白, 具有接头蛋白的活性,在细胞骨架与细胞外基... 详细信息

肝癌复发和转移是一个复杂的生物学过程,涉及多个基因和多种分子变化.研究表明,细胞的黏附动力学和形态改变在肿瘤的侵袭与转移中起重要作用.paxillin(桩蛋白)和p130Cas是两个黏着斑相关蛋白, 具有接头蛋白的活性,在细胞骨架与细胞外基质的黏附及细胞信号传导中起重要作用, 是很多不同癌蛋白的共同靶点[1-4].我们采用免疫组化和Western蛋白印迹的方法对原发性肝癌组织中paxillin和p130Cas蛋白表达水平进行分析,并探讨其与肝癌生物学行为的关系。

关键词： paxillin 肝癌复发 paxillin s蛋白细胞信号传导侵袭与转移蛋白表达水平癌生物学行为

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非小细胞肺癌组织中CD44v6和paxillin的表达及其与预后的关系

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四川大学学报（医学版） 2003年第3期34卷 484-485,488页

作者：左万里李海刚江门市中心医院内科江门529000 中山医科大学第二附属医院病理科

目的　研究细胞粘附分子在非小细胞肺癌组织中的表达及其与转移和预后的关系。方法　通过免疫组化的方法检测 79例非小肺癌组织中 CD44 v6和 paxillin表达并分析其与肿瘤的组织类型、分化程度、淋巴结转移与预后的关系。结果　肺癌组织... 详细信息

目的　研究细胞粘附分子在非小细胞肺癌组织中的表达及其与转移和预后的关系。方法　通过免疫组化的方法检测 79例非小肺癌组织中 CD44 v6和 paxillin表达并分析其与肿瘤的组织类型、分化程度、淋巴结转移与预后的关系。结果　肺癌组织中 CD44 v6和 paxillin的阳性率分别为 60 .8%和 3 4.1%。CD44 v6的表达与肺癌淋巴结转移呈正相关 ,与 2年生存率呈负相关 ( P均 paxillin的表达与肺癌淋巴结转移呈正相关 ( P均 <0 .0 5 )。结论　 CD44 v6和

关键词：非小细胞肺癌 CD44v6 paxillin 表达预后癌组织

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paxillin在涎腺腺样囊性癌中的表达

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口腔医学研究 2011年第12期27卷 1098-1100,1104页

作者：王姗赵尔杨方明施磊徐欣哈尔滨医科大学附第一临床医学院.口腔医学院口腔病理教研室黑龙江哈尔滨150001 哈尔滨医科大学研究生学院黑龙江高等护理专科学校

目的:探讨paxillin与涎腺腺样囊性癌(salivary adenoid cystic carcinoma,SACC)预后的关系。方法:采用免疫组化分析47例SACC的paxillin表达,并统计学分析paxillin与SACC病理学分型、临床分期、局部复发、远处转移和患者生存率的关系。结... 详细信息

目的:探讨paxillin与涎腺腺样囊性癌(salivary adenoid cystic carcinoma,SACC)预后的关系。方法:采用免疫组化分析47例SACC的paxillin表达,并统计学分析paxillin与SACC病理学分型、临床分期、局部复发、远处转移和患者生存率的关系。结果:paxillin表达与SACC病理学分型、局部复发和患者生存率无明显相关,而与临床分期和远处转移呈正相关关系(Ppaxillin的过度表达在SACC中常见,其过度表达与信号传导通路的改变可能与SACC的细胞增殖及癌转移相关,从而促进SACC的远处转移;paxillin可作为预后指标来预测SACC的远处转移,指导临床治疗。

关键词：涎腺肿瘤癌腺样囊性 paxillin 免疫组织化学

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