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A novel mutation in ror2 led to the loss of function of ror2 and inhibited the osteogenic differentiation capability of bone marrow mesenchymal stem cells(BMSCs)

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BIOCELL 2023年第7期47卷 1561-1569页

作者： WENQI CHEN XIAOYANG CHU YANG ZENG YOUSHENG YAN YIPENG WANG DONGLAN SUN DONGLIANG ZHANG JING ZHANG KAI YANG Prenatal Diagnosis Center Shijiazhuang Obstetrics and Gynecology HospitalKey Laboratory of Maternal and Fetal Medicine of Hebei ProvinceShijiazhuang050011China Department of Stomatology Fifth Medical Center of Chinese PLA General HospitalBeijing100039China Institute of Hematology Fifth Medical Center of Chinese PLA General HospitalBeijing100039China Prenatal Diagnosis Center Beijing Obstetrics and Gynecology HospitalBeijing Maternal and Child Health Care HospitalCapital Medical UniversityBeijing100026China Department of Orthodontics Beijing Stomatological HospitalCapital Medical University School of StomatologyCapital Medical UniversityBeijing100050China

Receptor tyrosine kinase-like orphan receptor 2(ror2)has a vital role in osteogenesis.However,the mechanism underlying the regulation of ror2 in osteogenic differentiation is still poorly comprehended.A previous study by our research group showed that a novel compound heterozygous ror2 variation accounted for the autosomal recessive Robinow syndrome(ARRS).This study attempted to explore the impact of the ror2:c.904C>T variant specifically on the osteogenic differentiation of BMSCs.Methods:Coimmunoprecipitation(CoIP)-western blotting was carried out to identify the interaction between ror2 and Wnt5a.Double-immunofluorescence staining was used for determining the expressions and co-localization of ror2 and Wnt5a in bone marrow mesenchymal stem cells(BMSCs).Western blot(WB)analysis and quantitative reverse transcription polymerase chain reaction(RT-qPCR)were conducted to identify the expression levels of ror2 in the BMSCs transfected with LV-shror2 or LV-ror2-c.904C>T.The alkaline phosphatase(ALP)activity was detected,and Alizarin Red S staining was done for evaluating the osteogenic differentiation of BMSCs.RT-qPCR was employed to identify the expression of the sphingomyelin synthase 1(SMS1)mRNA in the BMSCs transfected with LV-shror2 or LV-ror2-c.904C>T and the mRNA expression levels of Runt-related transcription factor 2(RUNX2),osteocalcin(OCN),and osteopontin(OPN).WB was performed to confirm the protein expressions of extracellular regulated protein kinases1(ERK),P-ERK,Smad family member1/5/8(Smad1/5/8),P-Smad1/5/8,P-P38,P38,RUNX2,OCN,and OPN in the BMSCs transfected with LV-shror2/LV-ror2-c.904C>T and sphingomyelin(SM).Results:The ror2:c.904C>T mutant altered the subcellular localization of the ror2 protein,which caused an impaired interaction between ror2 and Wnt5a.The depletion of ror2 restricted the osteogenic differentiation capability of BMSCs and downregulated the expression of SMS1.SM treatment could reverse the inhibition of osteoblastic differentiation in ror2-depleted BMSCs.Conclusion:The findings of

关键词： Bone marrow mesenchymal stem cells ror2 Wnt5a Osteogenic differentiation Sphingomyelin

Comparative study of ror2 and WNT5a expression in squamous/adenosquamous carcinoma and adenocarcinoma of the gallbladder

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World Journal of Gastroenterology 2017年第14期23卷 2601-2612页

作者： Zheng-Chun Wu Li Xiong Ling-Xiang Wang Xiong-Ying Miao Zi-Ru Liu Dai-Qiang Li Qiong Zou Kui-Jie Liu Hua Zhao Zhu-Lin Yang General Surgery Department Second Xiangya HospitalCentral South University Research Laboratory of Hepatobiliary Diseases Second Xiangya HospitalCentral South University Department of Pathology Second Xiangya HospitalCentral South University Department of Pathology Third Xiangya HospitalCentral South University

AIM To investigate the expression and clinical pathological significance of ror2 and WNT5 a in gallbladder squamous/adenosquamous carcinoma(SC/ASC) and adenocarcinoma(AC). METHODS En Vision immunohistochemistry was used to stain for ror2 and WNT5 a in 46 SC/ASC patients and 80 AC patients. RESULTS Poorly differentiated AC among AC patients aged > 45 years were significantly more frequent compared with SC/ASC patients, while tumors with a maximal diameter > 3 cm in the SC/ASC group were significantly more frequent compared with the AC group. Positive ror2 and WNT5 a expression was significantly lower in SC/ASC or AC with a maximal mass diameter ≤ 3 cm, a TNM stage of?Ⅰ?+ Ⅱ, no lymph node metastasis, no surrounding invasion, and radical resection than in patients with a maximal mass diameter > 3 cm, TNM stage Ⅳ, lymph node metastasis, surrounding invasion, and no resection. Positive ror2 expression in patients with highly differentiated SC/ASC was significantly lower than in patients with poorly differentiated SC/ASC. Positive ror2 and WNT5 a expression levels in highly differentiated AC were significantly lower than in poorly differentiated AC. Kaplan-Meier survival analysis showed that differentiation degree, maximal mass diameter, TNM stage, lymph node metastasis, surrounding invasion, surgical procedure and the ror2 and WNT5 a expression levels were closely related to average survival of SC/ASC or AC. The survival of SC/ASC or AC patients with positive expression of ror2 and WNT5 a was significantly shorter than that of patients with negative expression results. Cox multivariate analysis revealed that poor differentiation, a maximal diameter of the mass ≥ 3 cm, TNM stage Ⅲ or Ⅳ, lymph node metastasis, surrounding invasion, unresected surgery and positive ror2 or WNT5 a expression in the SC/ASC or AC patients were negatively correlated with the postoperative survival rate and positively correlated with mortality, which are risk factors and independent prognostic predictors. CONCLUSION SC/ASC or AC patients with

关键词：胆囊 adenosquamous 癌胆囊有鳞的癌 ror2 胆囊腺癌 WNT5a Immunohistochemistry

ror2 regulates the survival of murine osteosarcoma cells in lung capillaries

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Journal of Cancer Metastasis and Treatment 2020年第1期6卷 611-621页

作者： Diem Thi Phuong Tran Takahiro Kuchimaru Mongkol Pongsuchart Kha The Nguyen John Clyde Co Soriano Tetsuya Kadonosono Shinae Kizaka-Kondoh School of Life Science and Technology Tokyo Institute of TechnologyYokohama 226-8501Japan Center for Molecular Medicine Jichi Medical UniversityTochigi 329-0498Japan

Aim:Lung metastasis is a leading cause of death in patients with osteosarcoma(OS).No effective therapy exists that improves the five-year overall survival rate of OS patients with metastasis.Therefore,finding novel therapeutic targets will help develop new treatment strategies for OS patients with lung metastasis.Methods:Based on analysis of gene expression profiles between sublines of the Dunn OS LM8 cell line with high(LM8-H)and low(LM8-L)metastatic ability,we have identified Wnt signal-related genes that play an important role in lung metastasis of OS.Function of the genes was investigated by establishing sublines of gene knockout and assessing their metastatic ability using a mouse lung metastasis model.The molecular mechanism underlying the function of the genes was further investigated by in vitro experiments.Results:We have identified that receptor tyrosine kinase-like orphan receptor 2(ror2),a receptor of the non-canonical Wnt signaling pathway,was involved in OS cell survival in lung capillaries during metastasis.LM8-H knocked out of ror2(H/ror2-KO)significantly reduced lung metastasis by decreasing the viability in lung capillaries 48 h after intravenous injection.In vitro study revealed that ror2 increased anoikis resistance through AKT activation.Reconstitution of ror2 expression in H/ror2-KO cells restored their metastatic ability and viability in lung capillaries.Conclusion:The results demonstrate a novel ror2 function in OS lung metastasis and may inform new treatment strategies for OS patients.

关键词： Osteosarcoma lung metastasis cell survival lung capillary ror2

IFT20相关蛋白与肺腺癌患者预后的关联性研究

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IFT20相关蛋白与肺腺癌患者预后的关联性研究

作者：李连凤武汉科技大学

学位级别：硕士

目的:分析IFT20相关蛋白(intraflagellar transport 20,IFT20、cis-golgi protein GM130、Sonic Hedgehog,SHh和receptor tyrosine kinase-like orphan receptor 2,ror2蛋白)在肺腺癌患者癌组织及癌旁组织中的表达情况及其与患者临床病理特征及预后情况的关系,为早期诊断肺腺癌及评估其预后情况提供新的可能靶点。材料与方法:本研究随机选取了华中科技大学同济医学院附属同济医院2010年3月至2015年9月入院后行肺癌切除术的235例肺腺癌患者并随访至2018年6月。收集患者的基本信息、临床病理特征资料及术中切除的肺癌组织和癌旁组织。采用组织芯片和免疫组化技术检测分析IFT20相关蛋白在癌组织和癌旁组织中的表达情况,采用Image J软件对组织芯片进行IOD/Area和阳性细胞率评分,五名经专业培训的人员在不知患者临床资料的情况下根据染色的深浅对蛋白进行染色强度评分,其中未着色为0分;淡黄色为1分;棕黄色为2分;棕褐色为3分。符合正态分布的连续性变量采用独立样本的t检验或方差分析,不符合正态分布的连续性变量采用独立样本的Wilcoxon或Kruskal-Wallis秩和检验,分类变量采用卡方检验或Fisher确切概率法。Pearson相关或Spearman秩和相关分析IFT20相关蛋白之间的关系;多元Logistics回归分析IFT20相关蛋白的表达与肺腺癌临床病理特征之间的关系;多元Cox比例风险回归模型分析IFT20相关蛋白的表达与肺腺癌患者预后情况之间的关系。采用SPSS 26.0软件进行统计学分析,P3cm、肿瘤发生淋巴结或远处转移,以及肿瘤发展为II～Ⅳ期是影响肺腺癌患者预后的危险因素,其HR及95%CI分别为2.22(1.31,3.76),1.75(1.24,2.45),2.37(1.41,3.98),1.47(1.04,2.08),2.18(1.51,3.16)和2.23(1.45,3.42),而接受化疗/放疗是肺腺癌患者预后的保护因素,其HR及95%CI为0.58(0.43,0.80)。(2)肺腺癌患者癌组织中IFT20和SHh蛋白的表达明显高于癌旁组织;GM130和ror2蛋白的染色强度评分显著高于癌旁组织(P20、GM130、SHh和ror2蛋白之间的关系在癌组织和癌旁组织中均呈显著正相关(P20、GM130和SHh蛋白的表达分别与TNM分期、淋巴结转移、肿瘤细胞低分化类型和肿瘤大小等临床病理特征之间呈显著负相关,蛋白表达每增加一个单位,会降低肿瘤发展为III～Ⅳ期、发生淋巴结转移、肿瘤细胞转化为低分化类型和肿瘤发展为较大体积(>7cm)的风险,其OR及95%CI分别为0.65(0.44,0.96),0.63(0.40,0.99),0.71(0.53,0.94),0.97(0.94,0.99),0.64(0.41,0.98),0.97(0.95,0.99)和0.58(0.36,0.94)。而ror2蛋白的表达与肿瘤大小呈显著正相关,阳性细胞率每增加一个百分点,肿瘤发展为较大体积(>7cm)的风险增加8%,其OR及95%CI为1.08(1.01,1.16)。(4)SHh蛋白的表达对肺腺癌患者的预后具有保护作用,其HR及95%CI为0.60(0.46,0.80)。在肿瘤细胞为中高分化类型、肿瘤>7cm和发生远处转移的人群中,GM130和SHh蛋白的表达对于肺腺癌患者的预后具有保护作用(P20蛋白的表达会增加肿瘤发生远处转移患者死亡的风险(P3cm、肿瘤发生淋巴结转移或远处转移、肿瘤发展为II～Ⅳ期是影响肺腺癌患者预后的危险因素,而接受化疗/放疗是肺腺癌患者预后的保护因素。(2)IFT20、GM130和SHh蛋白的表达与肺腺癌临床病理特征之间呈显著负相关,蛋白表达水平越高,会降低肿瘤细胞向低分化类型转化和向癌症晚期发展的风险,有利于肺腺癌患者的预后。而ror2蛋白的表达与肿瘤大小呈显著正相关,蛋白表达越高,肿瘤向较大体积发展的风险越高。

关键词：肺腺癌 IFT20 GM130 SHh ror2 临床病理特征生存情况

Answering a century old riddle: brachydactyly type Al

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Cell Research 2004年第3期 179-187页

作者： Bo GAO, Lin HE Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China. Neuropsychiatric and Human Genetics Group, Bio-X Life Science Research Center, Shanghai Jiaotong University, Shanghai 200030, China. Department of Biochemistry, Faculty of Medicine, The University of Hong Kong, Hong Kong, China.

In 1903, Farabee analyzed the heredity of the human digital malformation, brachydactyly, the first recorded disorder of the autosomal dominant Mendelian trait. In 1951, Bell classified this type of brachydactyly as type A1 (BDA1). Over 100 cases from different ethnic groups have so far been reported. However, the real breakthrough in identifying the cause of BDA1 has only taken place in the last few years with the progress of the mapping and identification of one of the genes responsible for this disorder, thus providing an answer for a century old riddle. In this article, we attempt to review the current state of knowledge on the genetic features of BDA1 with its century-old history and signalling pathway of IHH, and also discuss genotype-phenotype correlation not only of BDA1, but also of all types of brachydactyly.

关键词： Brachydactyly IHH GDF5 ror2 BMPRIB.

circRNA_079813调控ror2和ERK1/2-MAPK信号通路对牙髓损伤大鼠成骨分化的影响

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广西医科大学学报 2023年第9期40卷 1455-1462页

作者：周洋张悦刘景新疆医科大学第五附属医院口腔科乌鲁木齐830011

目的:探讨circRNA_079813对牙髓损伤模型大鼠成骨分化的作用及其机制。方法:将70只SD大鼠随机分为7组,即假手术组、牙髓损伤组、pcDNA3.1空载体(pcDNA-null)+牙髓损伤组、过表达circRNA_079813(pcDNA-circ)+牙髓损伤组、pcDNA-circ+siRNA-NC+牙髓损伤组、pcDNA-circ+siRNA-ror2+牙髓损伤组、pcDNA-circ+PD98059(ERK1/2抑制剂)+牙髓损伤组,每组10只。术后3 d采用苏木精—伊红(HE)染色观察牙髓组织病理变化。检测各组碱性磷酸酶(ALP)活性,RTqPCR法检测circRNA_079813表达,western blotting法检测ror2、磷酸化(p-)ERK1/2、骨涎蛋白(BSP)、骨钙蛋白(OCN)、ALP、牙本质涎磷蛋白(DSPP)和牙本质基质蛋白-1(DMP-1)蛋白表达。结果:与假手术组比较,牙髓损伤组牙髓组织有明显炎症浸润和损伤特征,circRNA_079813表达下调(Pror2、p-ERK1/2、BSP、ALP、OCN、DSPP、DMP-1表达水平及ALP活性均升高(均Pror2表达后,与pcDNA-circ+siRNA-NC+牙髓损伤组比较,pcDNA-circ+siRNA-ror2+牙髓损伤组ror2、p-ERK1/2、BSP、ALP、OCN、DSPP、DMP-1表达及ALP活性均下降(均P2、ALP、BSP、OCN、DSPP、DMP-1表达及ALP活性均下降(均Pror2表达激活牙髓损伤大鼠ERK1/2-MAPK信号通路促进成骨分化。

关键词： circRNA_079813 成骨分化牙髓损伤 ror2 ERK1/2-MAPK信号通路

SiRNA诱导ror2表达下调抑制软骨肉瘤细胞的增殖、迁移和侵袭

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河北医药 2023年第10期45卷 1484-1487页

作者：黄建军石莺刘泉陈忠益曾国庆赵柏阳福建省厦门市海沧医院骨科 361026 厦门医学院病理学与病理生理学教研室

目的探讨小干扰RNA(SiRNA)抑制受体酪氨酸激酶样孤儿受体2(ror2)表达水平对软骨肉瘤细胞的增殖、迁移和侵袭的影响。方法将软骨肉瘤SW1353细胞分成空白对照组、NC组(转染NC-SiRNA)和ror2-SiRNA组(转染ror2-SiRNA)3组,分别给予相应的处理... 详细信息

目的探讨小干扰RNA(SiRNA)抑制受体酪氨酸激酶样孤儿受体2(ror2)表达水平对软骨肉瘤细胞的增殖、迁移和侵袭的影响。方法将软骨肉瘤SW1353细胞分成空白对照组、NC组(转染NC-SiRNA)和ror2-SiRNA组(转染ror2-SiRNA)3组,分别给予相应的处理后,用qRT-PCR和Western blot检测各组细胞中ror2 mRNA和蛋白的表达变化,CCK-8法检测各组细胞增殖,划痕实验及Transwell实验检测细胞迁移和侵袭能力的改变。结果ror2-SiRNA组ror2 mRNA和蛋白的表达水平明显低于空白对照组和NC组(P0.05);相同的作用时间,ror2-SiRNA组OD值明显低于空白对照组和NC组(P0.05)。细胞划痕实验结果显示,与NC组比较,ror2-SiRNA组肿瘤细胞的迁移距离明显变短,迁移能力明显受到抑制,差异有统计学意义(Pror2-SiRNA组的穿膜细胞数明显减少,侵袭能力明显减弱,差异有统计学意义(Pror2表达水平可显著抑制软骨肉瘤细胞的增殖、迁移和侵袭。

关键词：软骨肉瘤 ror2 SiRNA 迁移侵袭

ror2蛋白与肺腺癌患者临床特征及生存情况的关系

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华中科技大学学报（医学版） 2021年第4期50卷 509-515,521页

作者：李连凤周婷余其梅林慧王震宇秦灵芝孔宪荣廖伟武汉科技大学医学院公共卫生学院武汉430065 武汉科技大学医学院基础医学院武汉430065 华中科技大学同济医学院附属同济医院病理科武汉430030 华中科技大学同济医学院附属同济医院放射科武汉430030 武汉市第五医院呼吸与重症内科武汉430050

目的探讨受体酪氨酸激酶样孤核受体2(ror2)在肺腺癌组织中的表达情况,分析其与患者临床特征及生存情况的关系。方法采用组织芯片和免疫组织化学方法检测235例肺腺癌患者癌组织和癌旁组织中ror2蛋白的表达情况。采用SPSS 26.0软件进行统... 详细信息

目的探讨受体酪氨酸激酶样孤核受体2(ror2)在肺腺癌组织中的表达情况,分析其与患者临床特征及生存情况的关系。方法采用组织芯片和免疫组织化学方法检测235例肺腺癌患者癌组织和癌旁组织中ror2蛋白的表达情况。采用SPSS 26.0软件进行统计学分析。结果随访终点时存活56例,死亡179例,死亡病例中癌组织为低分化类型、肿瘤大小>3 cm、TNM分期为Ⅱ~Ⅳ期、以及发生淋巴结或远处转移的比例显著高于存活病例(均Pror2蛋白的相对表达量明显高于癌旁组织(Pror2蛋白阳性表达的患者中,当癌组织为低分化、TNM分期达到Ⅲ和Ⅳ期、发生淋巴结转移、细胞增殖指数Ki67≥50%时,ror2蛋白阳性或者强阳性表达率随着其表达程度的增强而增加(均Pror2高表达是肺腺癌患者发生死亡的危险因素,其OR及95%CI分别为3.42(1.11~10.56)、3.39(1.00~11.20)、3.37(1.03~10.97)和2.89(1.03~8.18)。结论ror2蛋白高表达可能是肺腺癌患者发生死亡的危险因素之一,检测肺组织中ror2的表达情况可能为早期干预和治疗肺腺癌提供新的思路。

关键词：肺腺癌 ror2 临床特征生存情况

GATA6、ror2和STAT3在膀胱移行细胞癌的表达及其意义

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重庆医科大学学报 2011年第5期36卷 535-538页

作者：张龙波苟欣何卫阳周青松重庆医科大学第一附属医院泌尿外科重庆400016

目的:探讨核转录因子GATA6、发育调节激酶ror2和信号转导因子STAT3在膀胱移行细胞癌中的表达及其临床意义。方法:采用免疫组织化学SP法检测72例膀胱移行细胞癌(Bladder transitional cell carcinoma,BTCC)组织中GATA6、ror2和STAT3的表... 详细信息

目的:探讨核转录因子GATA6、发育调节激酶ror2和信号转导因子STAT3在膀胱移行细胞癌中的表达及其临床意义。方法:采用免疫组织化学SP法检测72例膀胱移行细胞癌(Bladder transitional cell carcinoma,BTCC)组织中GATA6、ror2和STAT3的表达,并将结果与病理分级、临床分期、肿瘤复发等临床病理资料进行统计学分析。结果:GATA6、ror2和STAT3在BTCC的表达分别为:22.2%、61.1%、79.2%。不同病理分级和临床分期组间GATA6的表达差异无统计学意义(P>0.05),但复发组与未复发组间其表达差异有统计学意义(Pror2的表达差异有统计学意义(P0.05)。(2)ror2、STAT3在BTCC中的表达具有统计学相关性(rs=0.443,Pror2、STAT3无统计学相关。结论:STAT3、ror2和GATA6可能都参与了BTCC的进展;ror2与STAT3可能存在共同的信号通路;检测GATA6、ror2和STAT3可能对判断BTCC的进展和预后有一定价值。

关键词：膀胱移行细胞癌 GATA6 ror2 STAT3