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Roles of integrin β3 cytoplasmic tail in bidirectional signal transduction in a trans-dominant inhibition model

Frontiers of Medicine 2016年第3期10卷 311-319页

作者： Jiansong Huang Yulan Zhou Xiaoyu Su Yuanjing Lyu Lanlan Tao Xiaofeng Shi Ping Liu Zhangbiao Long Zheng Ruan Bing Xiao Wenda Xi Quansheng Zhou Jianhua Mao Xiaodong Xi State Key Laboratory of Medical Genomics Shanghai Institute of Hematology Collaborative Innovation Center of Hematology Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Institute of Hematology The First Affiliated Hospital School of Medicine Zhejiang University Hangzhou 310003 China Department of Hematology The First Affiliated Hospital of Nanchang University Nanchang 330006 China Department of Hematology Affiliated Hospital of Jiangsu University Zhenjiang 212001 China Department of Pediatrics Renji Hospital Shanghai Jiao Tong University School of Medicine Shanghai 200001 China Department of Hematology Peking Union Medical College Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing 100073 China Shanghai Institute of Hypertension Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Cyrus Tang Hematology Center Jiangsu Institute of Hematology 2011 Collaborative Innovation Center of Hematology Soochow University Suzhou 215123 China Sino-French Research Centre for Life Sciences and Genomics Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai 200025 China

We evaluated the roles of calpain cleavage-related mutations of the integrin β3 cytoplasmic tail in integrin αIIbβ3 bidirectional signaling using a trans-dominant inhibition model. Chimeric Tac-β3 proteins （i.e., Tac-β3, Tac-β3△741, Tac-β3△747, Tac-β3△754, Tac-β3△759, and Tac-β3ANITY） consisting of the extracellular and transmembrane domains of human IL-2 receptor （Tac） and the human integrin β3 cytoplasmic domain were stably expressed in the 123 CHO cells harboring human glycoprotein Ib-IX and wild-type integrin uIIbβ3. The different cells were assayed for stable adhesion and spreading on immobilized fibrinogen, and for binding soluble fibrinogen representing outside-in and inside-out signaling events, respectively. The chimeric protein Tac-β3 inhibited, and Tac-β3△NITY partially attenuated stable adhesion and spreading. Tac-β3, Tac-β3△759, Tac-β3ANITY, and Tac-β3△754, but not Tac-β3△747 or Tac-β3△741, impaired the soluble fibrinogen binding. Results indicated that the bidirectional signaling was significantly inhibited by Tac-β3△ and Tac-β3ANITY, albeit to a much lesser extent. Moreover, only inside-out signaling was impaired in the 123/Tac-β3△759 and 123/Tac-β3△754 cells in contrast to an intact bidirectional signaling in the 123/Tac-β3△747 and 123/Tac-β3△741 cells. In conclusion, the calpain cleavage of integrin β3 resulted in the regulatory effects on signaling by interrupting its interaction with cytoplasmic proteins rather than altering its conformation, and may thus regulate platelet function.

关键词： integrin β3 signal transduction tmns-dominant inhibition model

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Inhibitory Effects of Alpha-zearalenol on Angiotensin II-Induced integrin β_3 mRNA via Suppression of Nuclear Factor-κB

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Biomedical and Environmental Sciences 2005年第5期18卷 314-320页

作者： SU-MIN LI XIAO-MING WANG JIN QIU QIN SI HENG-YI GUO REN-YU SUN QI-XIA WU Department of Pathophysiology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences School of Basic Medicine Peking Union Medical College 5 Dong Dan San Tiao Beijing 100005 China

To investigate the effect of α-zearalenol on angiotensin If-induced β3 integrin mRNA expression in human umbilical vein endothelial cells （HUVECs）. Methods The mRNA level in integrin β3 was determined by reverse transcription-polymerase chain reaction. Endothelial NF-κB activity was determined by the luciferase activity assay of plasmid NF-κB-LUC. Results The angiotensin Ⅱ- induced β3 integrin mRNA expression was inhibited by α-zearalenol and 17β-estradiol （10nmol/L ^-1 μmol/L）, but not influenced by ICI 182, 780, a pure competitive antagonist for estrogen receptor or a nitric oxide inhibitor N^ω-Nitro-L-arginine methyl ester hydrochloride. Alpha-zearalenol and 17β-estradiol suppressed the angiotensin Ⅱ-induced activation of NF-κB in endothelial cells. Condusion Alpha-zearalenol inhibits angiotensin Ⅱ-induced integrin β3 mRNA expression by suppressing NF-κB activation in endothelial cells.

关键词： Alpha-zearalenol integrin β3 Endothelial cell NF-κB 17β-estradiol

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Correlation of integrin β3 mRNA and vascular endothelial growth factor protein expression profiles with the clinicopathological features and prognosis of gastric carcinoma

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World Journal of Gastroenterology 2008年第3期14卷 421-427页

作者： Shu-Guang Li Zai-Yuan Ye Zhong-Sheng zhao Hou-Quan Tao Yuan-Yu Wang Chun-Yu Niu Zhejiang Provincial People's Hospital Hangzhou 310014 Zhejiang Province China Zhejiang Chinese Medical University Hangzhou310053 Zhejiang Province China Hebei North University Zhangjiakou07500 Hebei Province China

AIM： To investigate integrin β3 mRNA and vascular endothelial growth factor （VEGF） protein expression in gastric carcinoma, and its correlation with microvascular density, growth-pattern, invasion, metastasis and prognosis. METHODS： In situ hybridization（ISH） of integrin β3 mRNA and immunohistochemistry of VEGF and CD34 protein were performed on samples from 118 patients with gastric cancer. RESULTS： The positive rate of integrin β3 mRNA in non-tumor gastric mucosa （20%） was significantly lower than that of the gastric cancer tissue （52.5%, X^2 = 10.20, P 〈 0.01）. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of integrin β3 mRNA were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of VEGF protein were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the mean MVD were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. It was found that the positive expression rate of integrin β3 mRNA was positively related to that of VEGF protein （P 〈 0.01） and MVD （P 〈 0.05）, meanwhile the positive expression rate of VEGF protein was positively related to MV

关键词： Stomach neoplasms integrin β3 Vascularendothelial growth factor Metastasis Prognosis

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Effects of Er'zhi Tiangui Granule (二至天癸颗粒) on Sequential Expressions of integrin β3 and Its Ligand Osteopontin in Mouse Endometrium during Controlled Ovarian Hyperstimulation

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Chinese Journal of Integrative Medicine 2012年第11期18卷 846-849页

作者：孙振高连方贾青孙金龙李婷婷郭颖张建伟张宁刘卉王利红 Reproductive Center the Affiliated Hospital of Shandong University of Traditional Chinese Medicine Basic Medicine Research Institution Shandong Academy of Medical Sciences Clinical College Shandong University of Traditional Chinese Medicine

To investigate the effects of ECzhi Tiangui Granule （二至天癸颗粒, ETG） on sequential expressions of integrin β 3 and its ligand osteopontin in the mouse endometrium during controlled ovarian hyperstimulation （COH） and implantation period. Methods： Seventy-five Mature female Kunming mice were randomly divided into 3 groups, a normal control group, a model group, and a treatment group administrated with ETG for 10 days, 25 in each group. After mated with male mice, every 5 mice were sacrified in each group at the 0, 2nd, 4th, 6th, and 8th days to take their endometrium. In-situ hybridization was used to detect the expressions of integrin β 3 and osteopontin in the endometrium. Results： mRNA expressions of integrin β 3 and osteopontin in the endometrium during implantation period showed similar time sequence rules in the treatment group to those in the normal control group; the peak values of them were a little lower in the treatment group than the normal control without significant differences. In the model group, integrin β 3 mRNA expression was higher at the 2nd day, obviously lower at the 4th and 6th days, and insignificantly lower at the 8th day; and osteopontin expression was remarkably lower at the 4th, 6th, and 8th days, compared with the normal control and the treatment groups （P〈0.05, P〈0.01）. Conclusions： COH might influence the sequential expressions of integrin β 3 and its ligand osteopontin, bring forward the integdn β 3 expression peak, impact on the cooperation of integrin β 3 and osteopontin, so as to damage the endometrial receptivity. ETG could regulate the sequential expressions of integrin β 3 and its ligand osteopontin to improve the mouse endometrial receptivity during COH.

关键词： Kidney-tonifying Er'zhi Tiangui Granule controlled ovarian hyperstimulation endometrialreceptivity integrin β3 osteopontin mouse

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integrin β3-associated microenvironment promotes invasion and angiogenesis in EGFRv III-expressing glioblastoma

Integrin β3-associated microenvironment promotes invasion a...

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第十届全国免疫学学术大会

作者： zhaoyu liu lei han yucui dong yanli tan yongsheng li manli zhao hui xie huanyu ju he wang yu zhao qifan zheng qixue wang jun su chuan fang tao jiang jiaren liu xia li chunsheng kang huan ren Harbin Medical University Tianjin Medical University General Hospital Hebei University Tianjin Neurological Institute The Oncology Hospital affiliated to Harbin Medical University The Hospital affiliated to Hebei University Beijing Tiantan Hospital Capital Medical University Boston Children’s Hospital and Harvard Medical School

Backgroud:Glioblastoma(GBM)is one of the most lethal brain tumors with a short survival *** of a variant type III mutant of the epidermal growth factor receptor(EGFRv III)renders tumor properties of the advanced stage,yet functional interactions of EGFRv III-expressing GBM and the tumor microenvironment,and targeted therapy remain unclear.

关键词： tumor microenvironment EGFRv III integrin β3 glioblastoma cilengitide

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乳腺癌中integrin β3通过调控PPARγ表达影响TAMs向M2极化的研究

乳腺癌中Integrin β3通过调控PPARγ表达影响TAMs向M2极化的研究

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作者：唐清南京大学

学位级别：硕士

细胞粘附分子整合素integrin β3广泛表达于多种肿瘤组织,具有促肿瘤侵袭、迁移以及血管生成的功能。临床数据显示,在乳腺癌中,integrin β3与患者的不良预后密切相关,显著影响患者无病生存期(DFS)和5年存活率。因此大量研究尝试靶向Int... 详细信息

细胞粘附分子整合素integrin β3广泛表达于多种肿瘤组织,具有促肿瘤侵袭、迁移以及血管生成的功能。临床数据显示,在乳腺癌中,integrin β3与患者的不良预后密切相关,显著影响患者无病生存期(DFS)和5年存活率。因此大量研究尝试靶向integrin β3以治疗肿瘤,并取得阶段性进展。其中integrin β3拮抗剂西仑吉肽(Cilengitide)在乳腺癌临床I期试验中展现了较好的疗效,其可通过抑制血管生成抑制肿瘤细胞生长;与紫杉醇联用,通过不同的肿瘤杀伤机制,发挥更强的治疗效果。上述报道显示,integrin β3在肿瘤临床治疗中具有较好的应用价值。但是,靶向integrin β3的治疗对肿瘤微环境影响的报道较少。integrinβ3是否可以通过影响肿瘤免疫微环境重塑而影响肿瘤进程,需要进一步了解其在肿瘤进程中的作用及其调控机制,这将有助于了解integrin β3抑制剂在肿瘤微环境中的作用,扩大其临床适用范围,为临床用药提供理论依据。肿瘤相关巨噬细胞(TAMs)是乳腺癌肿瘤免疫微环境中占比最大的免疫细胞,通常表现为M2-like表型,具有促肿瘤转移和介导免疫抑制的功能,其特征为高表达清道夫受体CD163,CD204及CD206;大量分泌免疫抑制性细胞因子,如:IL-10、TGF-β等;同时高表达精氨酸酶,调控精氨酸代谢产生多胺,在抑制T细胞的同时,促进乳腺癌细胞生长和增殖。因此,在乳腺癌中,TAMs大量浸润通常与不良临床预后密切相关。而乳腺癌中integrin β3对TAMs表型及功能的影响,同时阐明其中分子调控机制,对了解integrin β3在调控肿瘤免疫微环境中的作用具有重要意义。据此,我们结合细胞及动物模型,综合运用分子、细胞生物学手段,联合使用小分子抑制剂Cyclo(RGDyK)及CRISPR/Cas9基因编辑技术,深入探究integrin β3对乳腺癌TAMs功能的影响,阐释integrin β3调控巨噬细胞M2型极化的分子基础,为后续研究靶向integrin β3调控TAMs功能,重塑肿瘤免疫微环境而影响乳腺癌进展提供了扎实的实验基础。实验结果显示:1.在乳腺癌小鼠模型中,TAMs高表达integrin β3。2.进一步研究TAMs中integrin β3是否与促肿瘤的M2极化表型有关,在体外诱导TAMs模型中,加入Cyclo(RGDyK)处理,并不影响TAMs的促肿瘤相关基因,例如VEGF、MMP2等的表达,但M2表型相关基因:Arg1显著下调;IL-10部分降低;巨噬细胞抗原呈递能力增加。证明TAMs中integrin β3参与M2型极化有关。3.在IL-4诱导的M2型极化模型中,巨噬细胞integrin β3显著升高,Cyclo(RGDyK)处理后,M2表型相关基因:Arg1、CD206、Ym1、Fizz1等显著下调;巨噬细胞吞噬能力下降;抗原递呈能力提高。进一步验证巨噬细胞中integrin β3参与M2型极化有关。4.为了探讨巨噬细胞中integrin β3参与M2型极化的机制,我们敲除integrinβ3,发现integrin β3能够直接调控PPARy而不是STAT6信号介导巨噬细胞的M2型极化。*** β3还可调控PPARγ脂质合成相关功能,提示其可能通过影响脂质代谢而调控巨噬细胞表型。综上所述,本论文通过一系列体内、体外实验,确证了integrin β3在乳腺癌TAMs中高表达,并通过特异性调控PPARy而不是STAT6信号介导TAMs M2型极化。本研究探讨了 integrin β3在调控乳腺癌TAMs表型及功能中的作用及其机制,提示靶向TAMs中的integrin β3以及其下游效应分子可能成为一种新的基于免疫调控的乳腺癌治疗新策略。

关键词： integrin β3 肿瘤微环境 TAMs 巨噬细胞M2型极化 PPARγ

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miR-124-3p inhibits proliferation, migration and invasion by adjusting ITGB3 in gastric cancer cells

miR-124-3p inhibits proliferation, migration and invasion by...

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第一届中国临床分子诊断大会

作者：吴茜白浩钟慧钰温阳宋兴勃应斌武四川大学华西医院

OBJECTIVE: The aim of the present study was to investigate the underlying molecular mechanism by which miR-124-3 p suppresses gastric cancer cell in the proliferation, migration and invasion by adjusting ITGB3. MATERIALS AND METHODS: RT-PCR and western blot are used to detect the expression of miR-124-3 p, ITGB3 and their proteins intergrin β3 in three kinds of human gastric cancer cells MKN45, AGS, MGC-803 and normal human gastric epithelial cells GES-1, and further to identify the proliferation, migration and invasive capabilities of those cells. Select two kinds of GC cells with relatively high and low invasion and migration ability and divided into the blank control group, pEGFP-ITGB3 group, pEGFP-NC group, ITGB3 siRNA group, and control siRNA group. The wound healing, CCK-8 assay, transwell migration, and invasion assay was performed to test the effect of ITGB3 in cell proliferation, migration and invasion. Similarly, set the blank control group, miR-124-3 p mimics group, miR-NC group, miR-124-3 p inhibitor group and inhibitor-NC group, use the same method to detect the effect of mir-124-3 p in cell function, and RT-PCR and western blot are used to detect the expression of mir-124-3 p, ITGB3 and intergrin β3. RESULTS: The trend of ITGB3 expression level of four kinds of cell lines is MGC803 ＞ AGS ＞ MKN-45 ＞ GES-1 and the trend of miR-124-3 p expression level of cell lines is the opposite（p all ＜ 0.05）. CCK-8 assay, wound healing and transwell invasion and migration experiments showed that the cell lines with low expression of miR-124-3 p and high expression of ITGB3 gene as well as protein have high proliferation and invasion ability, vice versa. In GC cell lines MGC-803 and MKN45, the results of ITGB3 interference and overexpression experiment have shown that compared with control siRNA group, GC cell proliferation, invasion and metastasis ability has been restricted in ITGB3 siRNA group（ p ＜ 0.05）. Compared with p EGFP-NC group, GC cell proliferation, invasio

关键词： gastric cancer mir-124-3p ITGB3 integrin β3

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