Non-invasively differentiate non-alcoholic steatohepatitis by visualizing hepatic integrin αvβ3 expression with a targeted molecular imaging modality

BACKGROUND Non-invasive methods to diagnose non-alcoholic steatohepatitis(NASH),an inflammatory subtype of non-alcoholic fatty liver disease(NAFLD),are currently unavailable.AIM To develop an integrin αvβ3-targeted molecular imaging modality to differentiate NASH.METHODS Integrinαvβ3 expression was assessed in Human LO2 hepatocytes Scultured with palmitic and oleic acids(FFA).Hepatic integrinαvβ3 expression was analyzed in rabbits fed a high-fat diet(HFD)and in rats fed a high-fat,high-carbohydrate diet(HFCD).After synthesis,cyclic arginine-glycine-aspartic acid peptide(cRGD)was labeled with gadolinium(Gd)and used as a contrast agent in magnetic resonance imaging(MRI)performed on mice fed with HFCD.RESULTS Integrin αvβ3 was markedly expressed on FFA-cultured hepatocytes,unlike the control hepatocytes.Hepatic integrin αvβ3 expression significantly increased in both HFD-fed rabbits and HFCD-fed rats as simple fatty liver(FL)progressed to steatohepatitis.The distribution of integrinαvβ3 in the liver of NASH cases largely overlapped with albumin-positive staining areas.In comparison to mice with simple FL,the relative liver MRI-T1 signal value at 60 minutes post-injection of Gd-labeled cRGD was significantly increased in mice with steatohepatitis(P<0.05),showing a positive correlation with the NAFLD activity score(r=0.945;P<0.01).Hepatic integrin αvβ3 expression was significantly upregulated during NASH development,with hepatocytes being the primary cells expressing integrin αvβ3.CONCLUSION After using Gd-labeled cRGD as a tracer,NASH was successfully distinguished by visualizing hepatic integrin αvβ3 expression with MRI.

Implication of Expression of Osteopontin and Its Receptor Integrin αvβ3 in the Placenta in the Development of Preeclampsia

To investigate the expression of osteopontin （OPN） and its receptor integrin αvβ3 in the placental tissue from pregnant women complicated with preeclampsia, the expression of OPN and αvβ3 in the placenta of the pregnant women with preeclampsia and healthy pregnant women was detected by immunohistochemistry, Western blotting and RT-PCR. Our results showed that OPN and αvβ3 protein were expressed in the placenta from normal pregnant woman and those with preeclampsia. OPN was located in the placental syncytiotrophoblasts and the cytoplasm of capillary endothelial cells and integrin αvβ3 was mainly expressed on the surface of trophoblast cells. Expression of OPN and integrin αvβ3 in the placental tissue from preeclampsia subjects was significantly lower than that from the control group （P〈0.05）. Compared with the control group, expression of OPN in the placental tissue from preeclampsia group was significantly lower （P〈0.05） but there was no significant difference in the expression of αv and β3 between the preeclampsia group and the controls. It is concluded that OPN and its receptor integrin αvβ3 may be involved in the pathogenesis of preeclampsia.

Effects of Integrins and Integrin αvβ3 Inhibitor on Angiogenesis in Cerebral Ischemic Stroke

Summary： Integrins such as αvβ3, α5β31 play a key role in angiogenesis regulation, invasion and metastasis, inflammation, wound healing, etc. The up-regulation of integrin αvβ3 after cerebral ischemic stroke can promote angiogenesis, which in turn improves functional recovery. In addition, the integrin αvβ3 inhibitor can block the blood-brain barrier （BBB） leakage induced by vascular endothelial growth factor （VEGF） and also can reduce inflammatory reaction, decrease the deposition of fibrinogen. Other studies showed that integrin αvβ3 is not essential in revascularization. Therefore, the effect of integrin αvβ3 in the whole process of brain function recovery merits further study.

Effects of controlled superovulation on the expression of HOXA10,integrin αvβ3 and LIF in mice during peri-implantation

Objective:To observe the expression of endometrial homologous box gene A10(HOXA10),Integrinαvβ3 and leukemia suppressor factor(LIF)in peri-implantation mice,and to investigate the effect of controlled superovulation(COH)on endometrial receptivity in mice.Methods:After COH model preparation and blastocyst transplantation,kunming pure-bred mature mice were randomly divided into natural blastocyst+natural pseudocyst group,COH blastocyst+natural pseudocyst group,natural blastocyst+COH pseudocyst group,and COH blastocyst+COH pseudocyst group.The pregnancy rate and average number of beds in each group were observed on day 6 of conception.Western blot and RT-PCR were used to detect the expression of HOXA10,Integrinααvβ3,LIF protein and mRNA in the endometria of mice on day 5 of conception.Results:The pregnancy rate and average number of beds in the COH group were lower than those in the natural receptor group.The expression of HOXA10,Integrinααvβ3 and LIF in endometrium was significantly lower than that of the natural receptor mice(P<0.01).Conclusion:Superovulation drugs down-regulate the expression of HOXA10,Integrinαvβ3 and LIF in endometrium during the implantation window,reduce endometrial receptivity,and lead to low clinical pregnancy rate.

Integrin α_vβ_3靶向超声微泡的制备及体外寻靶实验

目的制备Integrinαvβ3靶向超声微泡,对其一般理化性质及体外寻靶能力进行检测。方法采用巯基-马来酰亚胺连接法制备携FITC标记iRGD肽的Integrinαvβ3靶向微泡(MBIntegrinαvβ3),并制备空白微泡(MBcontrol)。检测两组微泡的一般理化性质,在荧光显微镜下观察两组微泡荧光显像,流式细胞仪检测两组微泡FITC荧光含量。于光学显微镜及荧光显微镜下观察两组微泡与小鼠脑微血管内皮细胞(bEnd.3)的黏附情况。结果 1MBIntegrinαvβ3粒径为(0.84±0.26)μm,与MBcontrol粒径(0.79±0.19)μm差异无统计学意义(P>0.05);2荧光显微镜下观察,MBcontrol组微泡无荧光显示,MBIntegrinαvβ3组微泡表面显示绿色荧光;3流式细胞仪测得MBIntegrinαvβ3组微泡FITC荧光含量为98.4%,MBcontrol组微泡FITC荧光含量为2.5%;4体外寻靶实验显示,MBIntegrinαvβ3组微泡能聚集结合于bEnd.3细胞表面。结论成功制备了携iRGD肽的Integrinαvβ3靶向微泡,其具有良好的体外寻靶能力。

Effect of lentivirus-mediated integrin αVβ3-sh RNA on tumor growth of mice with lung cancer xenografts

Objective: To study the effect of lentivirus-mediated integrin αVβ3-sh RNA on tumor growth of mice with lung cancer xenograft. Methods: Lung cancer tissue, paracancer tissue and normal tissue were collected and integrin αVβ3 expression was detected; BALB/c nude mice were selected, divided into integrin αVβ3 knockdown group(KD group) and negative control group(NC group), and inoculated with cells stably infected by integrin αVβ3-sh RNA lentivirus and cells stably infected by negative control-sh RNA lentivirus respectively, the growth of tumor tissue was continuously observed, and the number of apoptosis cells as well as the expression of angiogenesis, apoptosis and invasion genes in tumor tissue were detected. Results: m RNA content and protein content of integrin αVβ3 in lung cancer tissue were significantly higher than those in paracancer tissue and normal tissue; increasing trend of tumor tissue volume of KD group was weaker than that of NC group, and tumor volume at various points in time of KD group was lower than that of NC group; m RNA contents and protein contents of VEGF, FGF, EGF, Bcl-2, MMP-9, MMP-12 and MMP-13 in tumor tissue of KD group were lower than those of NC group, and apoptosis index as well as m RNA content and protein content of Bax were higher than those of NC group. Conclusions: The expression of integrin αVβ3 increases in lung cancer tissue, and lentivirus-mediated integrin αVβ3-sh RNA can inhibit tumor growth of mice with lung cancer xenografts.

Ischemic preconditioning partially suppresses and postpones integrin α_Vβ_3 mRNA expression following transient global cerebral ischemia in C57BL/6 mice

Previous studies of integrin αvβ3 have focused on ischemic brain damage, although the role of integrin αvβ3 in ischemic preconditioning （IP） has rarely been reported. The present study analyzed the effects of IP on integrin αvβ3 mRNA expression following cerebral ischemia through the use of hematoxylin-eosin staining and real-time quantitative polymerase chain reaction techniques. Integrin avid3 mRNA expression in the ischemia group peaked at 24 hours after ischemia-reperfusion. In the IP ＋ ischemia group, integrin αvβ3 mRNA expression increased after 24 hours, but remained significantly less than the ischemia group, and expression continued to increase until 7 days after ischemiaJreperfusion. These results demonstrate that IP effectively attenuated upregulation of integrin αvβ3 mRNA expression at 24 hours after ischemia.

Effect of sirolimus on the expression of integrin αυβ3 mRNA in cardiac allograft in rats

Objective To investigate the effect of sirolimus ( SRL) on the expression of integrin αυβ3 mRNA in vascular smooth muscle cells of cardiac allografts in rats, and the possible mechanism of SRL in the prevention of cardiac allograft vasculopathy. Methods Heterotopic heart transplantation models were established Hearts

复发转移乳腺癌中FAK、Ezrin、Paxillin、Integrinβ3表达的意义及其与ERK/MAPK通路关系的研究

目的:探讨在复发转移乳腺癌中ERK/MAPK通路的激活情况以及FAK、Ezrin、Paxillin、Integrinβ3表达的意义及其与ERK/MAPK通路激活的关系。方法:收集1994～2006年住院接受根治手术的女性乳腺癌患者共100例,进行随访。分两组:实验组为术后复发转移者共46例,对照组为手术时间及发病年龄相近的术后未发生复发转移者共54例。常规病理蜡块切片,应用免疫组化、核酸原位杂交技术,检测ERK、FAK、Ezrin、Integrinβ3、Paxillin在两组中表达情况。结果:ERK、FAK、Ezrin、Integrinβ3在实验组中表达率均较对照组高(均P<0.05),实验组中ERK的表达分别与FAK、Ezrin呈正相关(均P<0.05)。多因素分析显示ERK、FAK、Ezrin是乳腺癌根治手术后复发转移的独立危险因素。结论:复发转移乳腺癌中,ERK/MAPK通路明显激活;Ezrin、FAK协同作用激活ERK/MAPK通路。FAK、Ezrin、Integrinβ3可作为乳腺癌术后复发转移的独立预测指标。

integrinβ3通过Src/ClC-3Cl-通道信号通路参与脑血管重构

脑血管重构是中风重要的危险因素之一,但发生机制不清楚.近期研究显示integrinβ3及ClC-3均参与血管增殖反应.本研究采用斑片钳单通道记录,结合分子生物学及免疫组化等技术在高血压脑血管重构的动物模型上,探讨integrinβ3与脑血管重构及其相关信号通路.研究发现,在自发性高血压大鼠,及双肾双夹高血压大鼠,以及血管紧张素Ⅱ（AngⅡ）引起的高血压小鼠上,integrinβ3的表达与脑血管重构呈良好正相关,敲除integrinβ3可抑制AngⅡ引起的脑血管平滑肌细胞的增殖,integrinβ3与Src及ClC-3有相互作用.integrinβ3能作用于ClC-3磷酸化的关键位点,286位酪氨酸磷酸化从而介导ClC-3的活性,ClC-3敲除可阻止integrinβ3的增殖作用及AngⅡ引起脑血管重构.研究结果表明,integrinβ3是通过Src/ClC-3信号通路参与脑血管重构.

整合素β3基因多态性蛋白质变异及其与疾病关系研究进展

整合素(ITG)β3基因位于17号染色体长臂2区1号3带(17q21.3),基因区间在47253827~47313743,全长59917 kb,编码区间在47253862~47310204,全长56343 kb,有15个外显子、14个内含子;编码1648个氨基酸残基,分子量为105 kDa。ITGβ3蛋白是一种二价离子依赖性细胞黏附分子,广泛存在于细胞膜中。其主要功能是介导细胞之间,细胞和细胞外基质之间的识别、连接和黏附,主导细胞内外跨膜接头蛋白连接的作用。近年来,关于ITG在细胞识别与黏附、细胞活化与分化、免疫反应、细胞信号传导、炎症反应、创伤修复与愈合、凝血与血栓形成、肿瘤形成与转移等方面发挥重要作用的研究取得许多进展,本文综述如下。

大肠癌中αvβ3 integrin表达与血管生成拟态的关系及分子机制的研究

目的:探讨大肠癌中是否存在血管生成拟态(VM),阐述VM存在的临床意义;分析VM与αvβ3 integrin表达的关系;研究大肠癌中αvβ3 integrin表达的临床意义及其与FAK、MMP-2、VEGF表达的相关性。方法:对227例大肠癌组织切片进行CD31免疫组织化学和PAS染色及αvβ3 integrin、FAK、VEGF、MMP-2免疫组织化学染色。结果:VM在大肠癌中阳性率为18.06%(41/227),αvβ3 integrin在大肠癌中阳性率为56.83%(129/227);VM阳性组、αvβ3 integrin阳性组生存时间均较阴性组短,差异均有统计学意义(P<0.05);VM阳性组的αvβ3 integrin阳性率较阴性组高,差异有统计学意义(P<0.05);αvβ3 integrin与FAK、VEGF、MMP-2表达呈正相关。结论:大肠癌中VM的存在、αvβ3 integrin的表达与肿瘤的不良预后有关;αvβ3 integrin可能通过与FAK、VEGF、MMP-2的相互作用,参与了大肠癌VM的形成。

整合素相关蛋白α2β与ανβ3在宫颈鳞癌组织中的表达及意义

目的 研究整合素相关蛋白α2β与ανβ3在宫颈鳞癌组织中的水平表达,并探讨其对疾病进展的意义。方法 收集2017年11月至2018年10月住院且手术切除的68例宫颈鳞癌患者的组织石蜡标本作为观察组,同时期住院行全子宫切除的52例子宫肌瘤患者的正常宫颈组织石蜡标本作为对照组。通过免疫组织化学法检测2组细胞膜上整合素相关蛋白α2β与ανβ3的表达水平,分析宫颈鳞状细胞癌的生物学行为与宫颈鳞状细胞癌细胞膜上α2β、ανβ3表达的关系。联合2个指标探讨整合素相关蛋白与宫颈癌临床分期及淋巴结转移的相关性。结果 观察组细胞膜上整合素α2β和ανβ3蛋白阳性表达率均显著高于对照组(P<0.05);2组细胞膜上整合素相关蛋白(α2β和ανβ3)表达在组织学分类、临床分期、淋巴结转移和浸润深度方面差异有统计学意义(P<0.05),在肿瘤大小方面差异无统计学意义(P>0.05)。α2β联合ανβ3阳性表达率与宫颈癌临床分期及淋巴结转移均呈正相关(r=0.941、0.931,P<0.05)。结论 整合素相关蛋白α2β与ανβ3表达在宫颈鳞状细胞癌组织患者标本中呈异常升高,其在疾病的发生发展中可能具有一定作用。同时,整合素相关蛋白与宫颈鳞状细胞癌的临床分期和淋巴结转移密切相关,可作为宫颈鳞癌局部浸润、远处转移的预测因素。

CXCR4通过integrinαVβ3调控头颈部鳞癌细胞迁移的机制研究

目的:研究C-X-C型趋化因子受体4(CXCR4)-整合素αVβ3(integrinαVβ3)介导头颈部鳞癌细胞迁移的分子生物学机制。方法:应用头颈部鳞癌细胞株HN-5,采用慢病毒转染方法构建CXCR4过表达的细胞株,用荧光显微镜观察基因表达情况;采用脂质体法将CXCR4-siRNA、integrinαVβ3-siRNA分别转染至HN-5细胞和CXCR4过表达的HN-5细胞中,Transwell小室实验检测HN-5细胞迁移能力,Western blot实验检测CXCR4、integrinαVβ3、基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶-2(MMP-2)和黏附斑激酶(FAK)-非受体型酪氨酸蛋白激酶(SRC)-细胞外信号调节激酶(ERK)信号通路相关蛋白的表达情况。结果:CXCR4过表达可上调HN-5细胞中integrinαVβ3表达水平,而抑制CXCR4表达可下调integrinαVβ3表达;CXCR4过表达可调控MMP-2和MMP-9促进癌细胞迁移,并激活FAK-SRC-ERK信号通路;而抑制integrinαVβ3表达可明显逆转CXCR4过表达对HN-5细胞的上述作用。结论:CXCR4过表达可通过integrinαVβ3促进头颈部鳞癌细胞迁移,其作用机制可能与激活FAK-SRC-ERK通路有关。

结直肠癌中αvβ3-integrin和MMP-11的表达及其临床意义

目的探讨结直肠癌中αvβ3-integrin和MMP-11的表达情况与各临床病理因素的关系,以及二者在结直肠癌中表达的相关性。方法采用原位杂交方法检测正常结直肠粘膜及结直肠癌组织中αvβ3-integrin和MMP-11的表达情况。结果结直肠癌组织中αvβ3-integrin和MMP-11的表达阳性率分别为57.5%和69.9%,与正常结直肠粘膜中的表达阳性率比较均有显著性差异(P<0.05);组织学分化差、淋巴结转移、临床分期高者αvβ3-integrin和MMP-11表达率增高,而且浸润较深者MMP-11表达率增高,与对应组比较均有显著性差异(P<0.05);在结直肠癌组织中,αvβ3-integrin表达和MMP-11表达呈正相关性(r=0.412,P<0.01)。结论提示αvβ3-integrin和MMP-11参与结直肠癌的发生、发展,可以作为结直肠癌的浸润和转移的预测指标。

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

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 MVD （P 〈 0.05）. The mean survival period in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥54.9/mm^2 was significantly shorter than that in patients with negative expression of integrin β3 mRNA （P 〈 0.05） and VEGF （P 〈 0.01）, and MVD 〈 54.9/mm^2 （P 〈 0.01）. Five-year survival rate in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥54.9/mm^2 was significantly lower than those with negative expression of integrin β3 mRNA （P 〈 0.05）, VEGF （P 〈 0.05）, and MVD 〈 54.9/mm^2 （P 〈 0.01）. CONCLUSION： Integrin β3 and VEGF expression can synergistically enhance tumor angiogenesis, and may play a crucial role in invasion and metastasis of gastric carcinoma. Therefore, they may be prognostic biomarkers and novel molecular therapeutic targets.

Tn-C、Integrin-α5β3在甲状腺滤泡状良、恶性肿瘤中的表达及意义

采用免疫组织化学法检测60份甲状腺滤泡状癌(FTC)组织(A组),60份甲状腺滤泡状腺瘤(FTA)组织(B组)及60份正常甲状腺组织(C组)标本中Tn-C、Integrin-α5β3的表达。结果A组Tn-C、integrin-α5β3阳性表达率显著高于B、C组(P均<0.05)。认为Tn-C、Integrin-α5β3可作为临床甲状腺滤泡状良、恶性肿瘤鉴别诊断的分子生化标记物,从而指导术式选择。

Ouabain inhibits anchorage-independent growth in human lung cancer cells via integrinαvβ3 reduction

Physiological effects of ouabain (Fig. 1A), a human endogenous hormone, have been intensively investigated nowadays.Recent studies demonstrate anticancer activity of ouabain in sensitization of apoptosis and suppression of migration in lung cancer cells (1,2)Focusing on metastatic process, the ability of cancer cells to grow in an anchorage-independent condition determines the success of cancer metastasis [3].

ANTI-HUMAN PLATELET TETRASPANIN(CD9)MONOCLONAL ANTIBODIES INDUCE PLATELET INTEGRIN αbβ3 ACTIVATION IN A Fc RECEPTOR INDEPENDENT FASHION

This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αbβ3 by the two mAbs. Results.HI117 and SJ9A4(10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets,suggesting that the two mAbs evoked activation of platelet integrin αbβ3.Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion.The anti platelet tetraspanin(CD9)mAbs,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors.Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process,with protein kinase C activation presumably being the common step of the three pathways.

基质金属蛋白酶-9(MMP-9)和整合素(Integrinβ3)在妊娠期高血压疾病患者胎盘组织中的表达及意义

目的研究基质金属蛋白酶-9(MMP-9)和整合素(Integrinβ3)在妊娠期高血压疾病患者胎盘组织中的表达情况,并探讨其与妊娠期高血压疾病发病的关系及两者的相关性。方法SP法分别检测Integrinβ3、MMP9在10例妊娠期高血压疾病患者及10例正常妊娠者胎盘组织中的表达。结果妊娠期高血压疾病组胎盘Integrinβ3表达明显高于正常妊娠组(P<0.01),妊娠期高血压疾病组胎盘MMP-9表达明显低于正常妊娠组(P<0.01)。结论本研究显示:与正常妊娠组比较妊娠期高血压疾病患者胎盘组织Integrinβ3表达明显增加、MMP9的表达明显减少,表明Integrinβ3、MMP9可能参与妊娠期高血压疾病的发生和发展。