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Expression and Clinical Significance of Semaphorin4D in Non-small Cell Lung Cancer and Its Impact on Malignant Behaviors of A549 Lung Cancer Cells 被引量:3
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作者 阮莎莎 李瑞超 +4 位作者 韩琦 刘静 李贵玲 宋颖秋 伍钢 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2014年第4期491-496,共6页
This study aimed to explore Semaphrin4D(Sema4D) expression and clinical significance in non-small cell lung cancer(NSCLC), and to define the roles and mechanisms of Sema4 D in regulating the malignant behaviors of... This study aimed to explore Semaphrin4D(Sema4D) expression and clinical significance in non-small cell lung cancer(NSCLC), and to define the roles and mechanisms of Sema4 D in regulating the malignant behaviors of A549 cells by small interfering RNA(siRNA). Firstly, immunohistochemistry revealed that Sema4 D was more frequently expressed in NSCLC than in lung benign lesion(P〈0.05) and its overexprssion was associated with low differentiation(P〈0.05), poor pTNM staging(P〈0.05) and occurrence of lymph node(LN) metastasis(P〈0.05). Endogenous Sema4 D expression was suppressed by Sema4 D siRNA in A549 cells overexpressing Sema4 D. Protein levels of Sema4 D, total Akt and p-Akt were examined by Western blotting. Cell proliferation, migration and invasion abilities were measured by MTT assay and Transwell assay respectively. Results showed that Sema4 D siRNA significantly suppressed phosphorylation of AKT in A549 cells, but it did not alter total AKT expression. In addition, efficient down-regulation of SemaD significantly inhibit cell proliferation(P〈0.05), migration(P〈0.05) and invasion(P〈0.05) in A549 cells. These findings suggest that Sema4 D might serve as a reliable tool for early prediction of NSCLC poor prognosis. Sema4 D could play an important role in promoting tumor proliferation, migration and metastasis in the NSCLC, by influencing the Akt protein phosphorylation. Inhibition of Sema4 D may be a useful approach for the treatment of NSCLC. 展开更多
关键词 semaphorin4d non-small cell lung cancer PROGNOSIS RNA interference PROLIFERATION migration INVASION
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Hypoxia-inducible factor-lαand semaphorin4D genes involved with tumor-associated macrophage-induced metastatic behavior and clinical significance in colon cancer 被引量:15
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作者 Mu Linjun Wang Jinshen Chen Yuezhi Li Leping Guo Xiaobo Zheng Sheng Jing Changqing 《Chinese Medical Journal》 SCIE CAS CSCD 2014年第20期3568-3575,共8页
Background Hypoxia promotes tumor angiogenesis and hypoxia-inducible factor-1 alpha (HIF-lg) plays a pivotal role in this process. Recently identified pro-angiogenic factor, semaphorin4D (Sema4D) also promotes ang... Background Hypoxia promotes tumor angiogenesis and hypoxia-inducible factor-1 alpha (HIF-lg) plays a pivotal role in this process. Recently identified pro-angiogenic factor, semaphorin4D (Sema4D) also promotes angiogenesis and enhances invasive proliferation in some tumors. Furthermore, tumor-associated macrophages (TAMs) can increase the expression of HIF-la and Sema4D in cancer cells and thus influence tumor growth and progression. The purpose of this study was to evaluate the effect of TAMs on the expression of Sema4D and HIF-la and the impact of biologic behavior in colon cancer cells. Methods Immunohistochemistry was used to analyze HIF-la and Sema4D expression in 86 curatively resected colon cancer samples and 52 normal colon tissues samples. The relationship between their expression and clinicopathological factors was analyzed. Furthermore, macrophage-tumor cell interactions, such as metastasis, angiogenesis, were also studied using in vitro co-culture systems. Statistical analysis was performed using SPSS 17.0 software (SPSS Inc., USA). Differences between two groups were analyzed with Student's t test. Results HIF-la (58%) and Sema4D (60%) were expressed at a significantly higher level in tumors than in normal tissues (P 〈0.01, for both). Furthermore, HIF-la and Sema4D expression was significantly correlated with lymphatic metastasis, specific histological types and TNM stages (P 〈0.05), but not with age and tumor size (P 〉0.05). Sema4D expression was correlated with that of HIF-la (r=0.567, P 〈0.01). TAMs markedly induced HIF-la and Sema4D expression in colon cancer calls and subsequently increased their migration and invasion. Conclusions HIF-la and Sema4D expression are closely related to lymphatic metastasis, specific histological types and TNM stages in colon cancer. Furthermore, TAMs promote migration and invasion of colon cancer cells and endothelial tube formation, possibly through up-regulation of HIF-la and Sema4D. 展开更多
关键词 colon cancer hypoxia-inducible factor-1 alpha semaphorin4d tumor-associated macrophages METASTASIS
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CD68和Sema 4D蛋白在胃癌组织中的表达相关性及临床意义 被引量:3
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作者 李军 孙玉宝 +5 位作者 王春雷 李志芳 李峰 王金申 靖昌庆 牟林军 《中国现代普通外科进展》 CAS 2016年第6期433-437,共5页
目的:比较肿瘤相关巨噬细胞(TAMs)标记抗原CD68和轴突导向因子4D(Sema4D)在胃癌组织和正常组织中的表达,探讨两者之间的相关性及临床意义。方法:采用免疫组织化学SP法检测86例胃癌组织和癌旁正常组织中CD68和Sema4D的表达,并分析其与临... 目的:比较肿瘤相关巨噬细胞(TAMs)标记抗原CD68和轴突导向因子4D(Sema4D)在胃癌组织和正常组织中的表达,探讨两者之间的相关性及临床意义。方法:采用免疫组织化学SP法检测86例胃癌组织和癌旁正常组织中CD68和Sema4D的表达,并分析其与临床病理特征的关系。结果:胃癌组织中CD68和Sema4D蛋白阳性表达率显著高于癌旁组织(72.1%vs23.3%和74.4%vs29.1%,均P<0.01)。CD68和Sema4D蛋白与胃癌组织分化类型、TNM分期和淋巴结转移明显相关(P<0.05),且两者表达呈正相关(r=0.467,P<0.01)。结论:胃癌组织中CD68和Sema4D表达明显增高,且呈显著正相关,两者的联合检测有助于预测胃癌的发展趋势。 展开更多
关键词 胃肿瘤 肿瘤相关巨噬细胞 CD 68 SEMA 4D
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Role of Inhibition of Osteogenesis Function by Sema4D/Plexin-B1 Signaling Pathway in Skeletal Fluorosis In Vitro 被引量:7
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作者 刘晓利 宋菁 +4 位作者 刘克俭 王文朋 许昌 张裕曾 刘芸 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2015年第5期712-715,共4页
Summary: Skeletal fluorosis is a chronically metabolic bone disease with extensive hyperostosis osteosclerosis caused by long time exposure to fluoride. Skeletal fluorosis brings about a series of abnormal changes of... Summary: Skeletal fluorosis is a chronically metabolic bone disease with extensive hyperostosis osteosclerosis caused by long time exposure to fluoride. Skeletal fluorosis brings about a series of abnormal changes of the extremity, such as joint pain, joint stiffness, bone deformity, etc. Differentiation and maturation of osteoblasts were regulated by osteoclasts via Sema4D/Plexin-B 1 signaling pathway. Furthermore, the differentiation and maturation of osteoclasts are conducted by osteoblasts via RANKL/RANK/OPG pathway. Both of these processes form a feedback circuit which is a key link in skeletal fluorosis. In this study, an osteoblast-osteoclast co-culture model in vitro was developed to illustrate the mechanism of skeletal fluorosis. With the increase of fluoride concentration, the expression level of Sema4D was decreased and TGF-β1 was increased continuously. OPG/RANKL mRNA level, however, increased gradually. On the basis of that, the inhibition of Sema4D/Plexin-B1/RhoA/ROCK signaling pathway caused by fluoride promoted the level of TGF-β1 and activated the proliferation of osteoblasts. In addition, osteroprotegerin (OPG) secreted by osteoblasts was up-regulated by fluoride. The competitive combination of OPG and RANKL was strengthened and the combination of RANKL and RANK was hindered. And then the differentiation and maturation of osteoclasts were inhibited, and bone absorption was weakened, leading to skeletal fluorosis. 展开更多
关键词 FLUORIDE semaphorin4d OSTEOBLAST OSTEOCLAST skeletal fluorosis
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三阴性乳腺癌中Sema4D与c-met表达及其与微淋巴管密度和各项临床病理特征的关系 被引量:1
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作者 蒋冰 黄波 +2 位作者 蔡存伟 刘骞 王正 《黑龙江医药》 CAS 2016年第2期231-234,共4页
目的:探讨三阴性乳腺癌Sema4D与c-met表达及其与微淋巴管密度(LMVD)和各项临床病理特征的关系。方法:采用免疫组化方法检测95例三阴性乳腺癌组织中Sema4D与c-met表达,检测微淋巴管密度(LMVD),依据不同临床病理特征分组并进行组间比较,... 目的:探讨三阴性乳腺癌Sema4D与c-met表达及其与微淋巴管密度(LMVD)和各项临床病理特征的关系。方法:采用免疫组化方法检测95例三阴性乳腺癌组织中Sema4D与c-met表达,检测微淋巴管密度(LMVD),依据不同临床病理特征分组并进行组间比较,同时以78例非三阴性乳腺癌及22例正常乳腺组织标本做对照。结果:三阴性乳腺癌组织中,Sema4D与c-met阳性表达率均明显高于其在非三阴性乳腺癌及在正常乳腺组织中的阳性表达率;Sema4D与c-met的表达具有正相关性(r=0.713,P<0.05)。三阴性乳腺癌组织中LMVD明显高于非三阴性乳腺癌及正常乳腺组织(P<0.05);Sema4D、c-met阳性表达组的LMVD明显高于阴性表达组(P<0.05);Sema4D表达在肿瘤大小、淋巴结转移数目及是否远处转移间比较,差异有显著性意义(P<0.05)。c-met表达在淋巴结转移数目及是否远处转移间比较,差异有显著性意义(P<0.05)。结论:Sema4D与c-met的高表达可能促进三阴性乳腺癌淋巴管的新生、淋巴道转移及远处转移,相关抑制剂的研究有可能成为预防和治疗三阴性乳腺癌淋巴道转移和远处转移的新方法。 展开更多
关键词 三阴性乳腺癌 轴突导向因子4D C-MET 微淋巴管密度
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肿瘤相关巨噬细胞和Sema4D与胃癌浸润转移的关系
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作者 李军 孙玉宝 +5 位作者 王春雷 李志芳 李峰 王金申 靖昌庆 牟林军 《中国现代普通外科进展》 CAS 2016年第7期511-515,共5页
目的:研究肿瘤相关巨噬细胞(TAMs)对人胃癌SGC-7901细胞系生物学功能的影响。方法 :采用佛波酯(PMA)、白细胞介素IL-4和IL-13体外诱导人M2型巨噬细胞,细胞免疫荧光鉴定TAMs。Transwell非接触式共培TAMs和胃癌SGC-7901细胞,侵袭实验、迁... 目的:研究肿瘤相关巨噬细胞(TAMs)对人胃癌SGC-7901细胞系生物学功能的影响。方法 :采用佛波酯(PMA)、白细胞介素IL-4和IL-13体外诱导人M2型巨噬细胞,细胞免疫荧光鉴定TAMs。Transwell非接触式共培TAMs和胃癌SGC-7901细胞,侵袭实验、迁移实验检测肿瘤相关巨噬细胞(TAMs)对胃癌SGC-7901细胞侵袭和迁移的影响,酶联免疫吸附法(ELISA)检测实验组和空白对照组胃癌SGC-7901细胞上清中分泌型Sema4D的变化。结果:细胞免疫荧光鉴定M2型巨噬细胞诱导成功;在Transwell共培养体系中,TAMs共培养的胃癌SGC-7901细胞形态学发生改变;Transwell侵袭实验、迁移实验表明,细胞侵袭转移力增强(P<0.01)。与TAMs共培养的胃癌SGC-7901细胞的上清液分泌型Sema4D蛋白明显增多,较空白对照组差异有统计学意义(1224.13±29.43比637.15±33.84,P<0.01)。结论 :TAMs可促进胃癌细胞的浸润转移,其原因可能与分泌型Se m a4D蛋白的表达上调有关。 展开更多
关键词 胃肿瘤·肿瘤相关巨噬细胞·S EM a4D·浸润转移
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沉默轴突导向蛋白4D基因对卵巢癌SKOV3细胞恶性生物学行为的影响 被引量:6
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作者 陈颖 张磊 +2 位作者 吴卉娟 刘文欣 郝权 《中华肿瘤杂志》 CAS CSCD 北大核心 2011年第5期324-330,共7页
摘要:目的 探讨沉默轴突导向蛋白4D(Sema4D)基因对卵巢癌SKOV3细胞增殖、侵袭、转移的影响,并验证Sema4D干扰RNA对人卵巢癌裸鼠移植瘤生长的抑制作用.方法 针对Sema4D基因构建短发夹RNA(shRNA)重组载体pshRNA Sema4D-A、pshRNASema... 摘要:目的 探讨沉默轴突导向蛋白4D(Sema4D)基因对卵巢癌SKOV3细胞增殖、侵袭、转移的影响,并验证Sema4D干扰RNA对人卵巢癌裸鼠移植瘤生长的抑制作用.方法 针对Sema4D基因构建短发夹RNA(shRNA)重组载体pshRNA Sema4D-A、pshRNASema4D-B和pshRNASema4D-C质粒,并行酶切鉴定和测序分析.采用逆转录聚合酶链反应(RT-PCR)法筛选RNA干扰效果最佳的重组载体,通过脂质体介导转染卵巢癌SKOV3细胞(重组载体组),并以空载体组(转染空载体pcDNA3.1质粒)和未转染组(未作任何处理)为对照.采用RT-PCR法和Westem blot法分别检测转染前后SKOV3细胞中Sema4D mRNA和蛋白表达,并榆测转染前后SKO3细胞的增殖、侵袭和转移能力.建立人卵巢癌裸鼠移植瘤模型,将满足成瘤标准的15只裸鼠随机分为pshRNASema4D组(重组载体组)、空载体组和未转染组,每组5只,分别向瘤块内注射pshRNASema4D-B(50μg/次)、空载体(50μg/次)和磷酸盐缓冲液(PBS,100μl/次),每3 d注射1次,共3周,观察移植瘤生长情况.结果 酶切鉴定和测序分析证实,靶向Sema4D的3个ShRNA重组载体pshRNASema4D-A、pshRNASema4D-B和pshRNASema4D-C质粒构建成功.RT-PCR法筛选显示,重组载体pshRNASema4D-B质粒干扰效果最佳.重组载体组转染24、48和72 h时,SKOV3细胞中Sema4DmRNA的相对表达水平分别为0.401.4±0.051、0.120±0.035和0.014±0.015,重组载体组转染72 h时的Sema4D mRNA相对表达水平明显低于空载体组(0.521±0.019)和未转染组(0.536±0.040,P〈0.05).Western blot法检测显示,重组载体组转染24、48和72 h时,细胞中Sema4D蛋白的相对表达水平分别为0.196±0.023、0.074±0.015和0.040±0.014,重组载体组转染72 h时的Sema4D蛋门相对表达水平明显低于空载体组(0.275±0.009)和未转染组(0.282±0.015,P〈0.05).与空载体组和未转染组比较,重组载体组细胞增殖、侵袭和迁移能力受到抑制(P〈0.05).注射pehRNASema4D-B后,裸鼠移植瘤生长缓慢,瘤块重量较空载体组和未转染组明显降低(P〈0.05).结论 RNA干扰技术能成功沉默SKOV3细胞中Sema4D基因的表达,通过下调Sema4D基因的表达呵抑制卵巢癌细胞增殖、侵袭及转移,Sema4D干扰RNA能抑制人卵巢癌裸鼠移植瘤的生长,Sema4D基因可能成为人卵巢癌基因靶向沉默治疗的重要候选基因之一. 展开更多
关键词 卵巢肿瘤 semaphorin4d RNA干扰 细胞增殖 侵袭 转移
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职业性氟接触人群氟负荷与血清中可溶性Sema4D水平的相关性研究 被引量:1
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作者 涂俊 刘克俭 +4 位作者 旷聃 金彦 刘富强 张裕曾 刘芸 《工业卫生与职业病》 CAS 2017年第6期438-441,共4页
目的检测职业性氟接触人群血清中可溶性Sema4D(sSema4D)水平,探讨sSema4D与氟负荷的相关性。方法选取湖北省某铝业公司91名氟接触工人和54名行政职员为研究对象。应用问卷法调查其人口统计学特征。采用离子选择电极法测定尿氟。采用酶... 目的检测职业性氟接触人群血清中可溶性Sema4D(sSema4D)水平,探讨sSema4D与氟负荷的相关性。方法选取湖北省某铝业公司91名氟接触工人和54名行政职员为研究对象。应用问卷法调查其人口统计学特征。采用离子选择电极法测定尿氟。采用酶联免疫吸附试验(ELISA)法检测sSema4D水平。应用SPSS 17.0软件对数据进行统计分析。结果氟接触工人无氟性骨损伤组(APW-FBI)的尿氟(UF)水平(4.83±2.62 mg/L)高于对照组(1.30±0.11 mg/L)(P<0.01),氟接触工人有氟性骨损伤组(APW+FBI)的UF水平(15.0±3.35 mg/L)高于APW-FBI组(P<0.01);APW-FBI组的sSema4D水平(2.99±0.30 ng/mL)低于对照组(5.84±0.06 ng/mL)(P<0.01),APW+FBI组的sSema4D水平(1.90±0.24 ng/mL)低于APW-FBI组(P<0.01);UF与sSema4D呈负相关(r=-0.76,P<0.01)。结论氟接触工人(尤其氟性骨损伤)sSema4D水平较低,且与UF水平呈负相关。 展开更多
关键词 损伤 Sema4D
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Sema4D/Plexin-B1在氟性骨损伤发生中作用的实验研究 被引量:2
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作者 金彦 刘富强 +2 位作者 刘克俭 刘芸 张裕曾 《工业卫生与职业病》 CAS 2017年第6期442-445,共4页
目的探讨信号素4D(Sema4D)及受体丛状蛋白B1(Plexin-B1)在氟性骨损伤发生中的作用。方法 SPF级SD大鼠48只,随机分为1个对照组和3个染氟组,采用氟化钠(NaF)溶液饮水染毒,3个染氟组NaF染毒浓度分别为1.6、16和32 mg/kg。在染氟第0天、第1... 目的探讨信号素4D(Sema4D)及受体丛状蛋白B1(Plexin-B1)在氟性骨损伤发生中的作用。方法 SPF级SD大鼠48只,随机分为1个对照组和3个染氟组,采用氟化钠(NaF)溶液饮水染毒,3个染氟组NaF染毒浓度分别为1.6、16和32 mg/kg。在染氟第0天、第15天、第30天和第90天时,进行大鼠尾动脉取血,分别测定各时段血氟(微量电极法)、ALP(全自动生化分析仪法)和Sema4D/Plexin-B1(ELISA法)。结果染氟30 d后,3个染氟剂量组大鼠均发生氟斑牙,表现为下切牙色泽度差,表面棕、白色带增宽,横纹不规则甚至消失,并可见牙齿缺损。大鼠血氟与ALP随染毒剂量增加和时间延长呈升高趋势,与对照组比较,差异有统计学意义(P<0.01)。大鼠Sema4D含量在染氟第15天开始降低,组间比较,差异有统计学意义(P<0.05)。染氟第30天,高剂量组(12.20±0.73 ng/1L)和中剂量组(13.70±0.68 ng/L)大鼠Sema4D含量下降明显,与对照组(19.20±0.68 ng/L)比较差异有统计学意义(P<0.01)。染氟第90天,高、中、低剂量组大鼠Sema4D含量分别为(11.14±0.66)、(11.73±0.68)和(13.72±0.71)ng/L,与对照组(19.87±0.69 ng/L)比较明显降低(P<0.01)。大鼠Plexin-B1含量也随着染氟时间的延长而有所下降,且高、中2个剂量组大鼠Plexin-B1含量下降较为明显,与对照组比较,差异有统计学意义(P<0.01)。结论染氟组大鼠血清中Sema4D和Plexin-B1表达量明显降低,大鼠氟性骨损伤的发生可能与Sema4D/Plexin-B1表达水平有关。 展开更多
关键词 信号素4D 大鼠 氟性骨损伤
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