钙池操纵性钙内流在非小细胞肺癌中的作用

目的探讨钙池操纵性钙通道（store-operated calcium channel,SOCC）介导的钙池操纵性钙内流（storeoperated calcium entry,SOCE）在非小细胞肺癌中的作用。方法应用实时定量PCR方法和Western blot方法检测非小细胞肺癌组织和A549肺腺癌细胞中SOCC的mRNA和蛋白表达。应用Fura-2Am荧光素方法检测A549细胞的SOCE;经过不同浓度SOCE抑制剂处理A549细胞后,应用WST-1比色法和Transwell方法检测A549细胞的增殖和侵袭能力改变。结果在非小细胞肺癌组织和A549细胞中有STIM1、ORAI1～3的mRNA和STIM1、ORAI1的蛋白表达,分化差的肿瘤组织比分化好的高表达ORAI1 mRNA（P=0.028）;A549细胞中存在SOCE,并且SOCE抑制剂SKF-96365或Ni Cl2可抑制A549细胞的增殖和侵袭。结论非小细胞肺癌组织中表达STIM1和ORAI1～3。A549肺腺癌细胞存在SOCE,并且由SOCC介导的SOCE在肺癌细胞生长和转移过程中起着重要的作用。     

钙释放激活钙通道调节分子1促进结肠癌细胞系SW480的迁移和侵袭

  目的  探讨钙释放激活钙通道调节分子1（calcium release-activated calcium channel modulator 1,ORAI1）对SW480迁移和侵袭的影响及其机制。  方法  以ORAI1干扰慢病毒感染SW480细胞,用RT-qPCR和Western blot检测细胞中ORAI1mRNA和蛋白的表达,Transwell小室、黏附实验、血管形成及拟态分别检测细胞侵袭、迁移能力,同、异种细胞间黏附及血管生成能力,激光共聚焦显微镜检测细胞钙内流（store-operated Ca2+ entry,SOCE）,Western blot法检测细胞中ERK1/2、p-ERK1/2、MMP-2、VEGF和E-cadherin蛋白的表达。  结果  SW480转染ORAI1干扰慢病毒72h后,可见明显的荧光表达;较空病毒组和（或）对照组,干扰组ORAI1的表达降低（P < 0.01）;侵袭和迁移能力减弱（P < 0.01）;同种黏附能力增强（P < 0.05）;异种黏附能力减弱（P < 0.05）;血管生成能力减弱（P < 0.01）;SOCE内流峰值降低（P < 0.05）;p-ERK1/2、MMP-2和VEGF的表达降低（P < 0.01）、E-cadherin表达增加（P < 0.01）。  结论  ORAI1可以促进SW480的迁移和侵袭,其机制可能与SOCE增加有关。      

Ebp1在肿瘤增殖和侵袭中的研究进展

肿瘤细胞的恶性增殖和浸润转移是影响肿瘤发展和治疗效果的重要因素,了解调控增殖和转移的影响因子及其分子机制对肿瘤早期的诊断和临床治疗意义重大。本文系统总结了增殖相关蛋白PA2G4（proliferation associated 2G4）家族成员Ebp1在多种肿瘤中的表达情况及其对细胞周期、细胞凋亡和分化、细胞迁移等方面的影响,并归纳了Ebp1影响肿瘤增殖和迁移的分子机制。Ebp在多种肿瘤中表达降低且与肿瘤的增殖和迁移呈负相关,并主要通过抑制周期相关蛋白的转录、调控影响肿瘤发展的信号通路。      

趋化因子CXCL12及其受体CXCR4介导乳腺癌转移分子机制

趋化因子CXCL12及其受体CXCR4之间的相互作用在乳腺癌转移中起着枢轴作用.新近研究发现,CXCR4-CXCL12介导的乳腺癌转移与缺氧诱导因子-1(HIF-1)、Her-2、核因子-κB(NFκB)、尿激酶型纤维蛋白酶原激活剂受体(uPAR)等分子密切相关,明确CXCL12-CXCR4介导的乳腺癌转移分子机制,进而逐个阻断上游分子对二者的调控作用,开发出靶点明确、作用特异的抗转移药物将使乳腺癌患者获益.     

VEGFC/D －VEGFR3／NRP2轴通路相关分子在肿瘤及其淋巴道转移时的变化与意义

本文综述 VEGFC ／D －VEGFR3／NRP2轴通路相关分子 Furin －like 酶、CNTN1、Prox1、LYVE －1、Podoplanin、SOX18、SDF1、CXCR4等的结构功能,在不同部位、不同类型、不同发展阶段的肿瘤及其淋巴道转移过程中的变化规律和细胞内外信号系统转导、执行、合成等网络调控特点以及肿瘤细胞－淋巴管内皮细胞相互作用及其生物学行为影响,特别是在肿瘤发生发展和淋巴道转移分子机制中的意义。     

基因表达芯片分析筛选胃癌转移相关基因

目的:筛选高侵袭型胃癌转移相关基因,探讨其在胃癌发生发展中可能通路的分子水平机制。方法:分别以年龄相近、性别相同的3例肝转移胃癌患者标本的转移灶和原发灶为实验组和对照组做高通量cDNA表达谱芯片,初选3张芯片共同上、下调基因;利用pathway V1.0分析软件寻找初筛基因中存在多种可能影响胃癌发生发展及其转归的分子通路;通过RT-PCR测定数例转移胃癌病例原发和转移灶中相关基因的mRNA转录水平。结果:通路分析筛选出来的上调基因CLDN1和下调基因UGT1A10在6组病例中经RT-PCR定性检测全部与分析结果相符。结论:pathway在芯片初选相关基因的基础上发现与疾病相关的分子机制通路及相关基因,为研究胃癌转移基因提供依据和参考。     

乙酰肝素酶与肿瘤转移研究进展

乙酰肝素酶是目前发现的哺乳动物细胞中唯一能切割细胞外基质中硫酸肝素蛋白多糖侧链的内源性糖苷酶。本文对乙酰肝素酶的结构、功能、分子特性、基因定位、核苷酸序列及其对血管生成的影响,在正常组织、肿瘤组织及转移癌组织中的表达和该酶参与肿瘤转移的机制等方面进行了综述。对用筛选乙酰肝素酶抑制剂的方法、寻找抗肿瘤新药的最新进展做了概述,旨在为肿瘤转移机制研究、寻找治疗方法提供一些线索。     

WISP-1/NGX6和G3BP/TIP30基因表达对肿瘤细胞增殖转移的影响

肿瘤的恶变过程包括细胞增生、DNA过度复制、细胞周期功能紊乱、细胞永生化、逃逸凋亡、血管增生及转移浸润等一系列过程.相应的分子机制为癌基因的激活、抑癌基因的失活、修复相关基因的功能缺失、凋亡机制缺失、信号转导调控机制紊乱及浸润转移相关分子事件等,而在肿瘤恶变的一系列分子机制中,细胞增殖凋亡、侵袭转移日益被重视.现全文就WISP- 1/NGX6、G3BP/TIP30两组基因表达对肿瘤细胞增殖转移的影响作一综述.     

MIIP通过HDAC6参与肿瘤侵袭转移的研究进展

迁移侵袭抑制蛋白（ migration and invasion inhibitory protein,MIIP）,是近年来研究发现的潜在肿瘤转移抑制基因,定位于1p36,该区域在多种肿瘤中缺失。MIIP的定位提示其可能在肿瘤侵袭、转移中发挥作用,并且已有研究报道MIIP可抑制胶质瘤及乳腺癌的侵袭转移。本文就MIIP通过HDAC6在肿瘤侵袭转移中的作用及其分子机制作一综述。     

恶液质与肿瘤转移的相关分子机制

恶液质是一种骨骼肌量进行性下降的代谢综合征,伴随或不伴随脂肪量减少,在晚期肿瘤患者中普遍存在。恶 液质降低抗肿瘤治疗疗效,增加肿瘤放化疗不良反应,增加肿瘤患者死亡率,是预后不良的重要因素之一。恶液质促进肿 瘤的侵袭和转移,其机制尚不明确,可能与恶液质患者中存在的炎性反应、乏氧状态、瘦素水平下降、促血管生成因子释 放等有关。肿瘤恶液质的病理生理机制错综复杂,普遍认为肿瘤转移的患者,肿瘤负荷增加,患者发生恶液质的风险增加, 且其治疗效果欠佳,目前仍缺乏标准的治疗方案。进一步明确恶液质的病理生理特点及其促进肿瘤转移的机制,有助于指 导早期营养筛查和营养评估,明确恶液质的诊断,并对恶液质进行分期和分级管理,有效地预防和治疗恶液质,进而减少 肿瘤转移,改善患者生活质量及预后。本文通过探讨恶液质的病理生理特点及其促进肿瘤转移的相关分子机制,综述恶液 质与肿瘤转移的相关分子机制,为恶液质及肿瘤转移的治疗提供重要依据。     

Blockade of store-operated Ca entry inhibits hepatocarcinoma cell migration and invasion by regulating focal adhesion turnover

Store-operated Ca²⁺ entry (SOCE) is a main Ca²⁺ influx pathway controlling the intracellular Ca²⁺ concentration in normal hepatocytes and hepatocellular carcinoma (HCC) cells. Ca²⁺ influx has been demonstrated to be involved in liver oncogenesis. Stromal interacting molecule (STIM) 1 acts as a sensor for the level of Ca²⁺ stored in the endoplasmic reticulum, and Orai1 protein constitutes the pore-forming subunit of the store-operated channels. Recently, STIM1 and Orai1 were found critical for breast tumor cell migration and metastasis. However, the effects of Ca²⁺ influx pathway on migration and metastasis have not been studied in hepatocellular carcinoma. Here, we found that STIM1 had a higher expression in hepatoma tissues than in precancerous tissues of the same patients. In general, STIM expression is elevated in HCC cell lines compared to a normal hepatocyte cell line. HCC-LM3 cell, which has a higher migration ability, expresses five times higher level of STIM than other HCC cell lines. STIM1 could then be explored as a prognostic marker to screen liver cancer patients with high metastatic potential. Inhibition of SOCE and STIM1 enhance focal adhesions and decrease the focal adhesion turnover, suggesting the therapeutic potential of SOCE and STIM1 as new molecular targets for metastatic HCC.     

Inhibition of store-operated Ca entry suppresses EGF-induced migration and eliminates extravasation from vasculature in nasopharyngeal carcinoma cell

Store-operated Ca²⁺ entry (SOCE) mediates Ca²⁺ responses evoked by extracellular signaling molecules to promote increases in cytosolic Ca²⁺, thereby triggering downstream signal transduction. Here we demonstrated that either the pharmacological blockage of Ca²⁺ influx through SOCE or the knockdown of Orai1, a key molecule of SOCE, suppressed the epidermal growth factor-induced migration by disturbing Ca²⁺ signaling in nasopharyngeal carcinoma (NPC) cell. Furthermore, Orai1 depletion led to a delayed cell attachment to the extracellular matrix surface in vitro and eliminated the extravasation of microinjected cells from vasculature in a zebrafish hematogenous metastasis model. Our findings thus indicate that SOCE acts as a predominant Ca²⁺ signaling involved in NPC cell metastasis, and may serve as a candidate target for anti-metastasis therapy in NPC.     

SOCE and cancer: Recent progress and new perspectives

Ca²⁺ acts as a universal and versatile second messenger in the regulation of a myriad of biological processes, including cell proliferation, differentiation, migration and apoptosis. Store‐operated Ca²⁺ entry (SOCE) mediated by ORAI and the stromal interaction molecule (STIM) constitutes one of the major routes of calcium entry in nonexcitable cells, in which the depletion of intracellular Ca²⁺ stores triggers activation of the endoplasmic reticulum (ER)‐resident Ca²⁺ sensor protein STIM to gate and open the ORAI Ca²⁺ channels in the plasma membrane (PM). Accumulating evidence indicates that SOCE plays critical roles in cancer cell proliferation, metastasis and tumor neovascularization, as well as in antitumor immunity. We summarize herein the recent advances in our understanding of the function of SOCE in various types of tumor cells, vascular endothelial cells and cells of the immune system. Finally, the therapeutic potential of SOCE inhibitors in the treatment of cancer is also discussed.     

Store-operated Ca2+ entry regulates glioma cell migration and invasion via modulation of Pyk2 phosphorylation

Background

The ubiquitous second messenger Ca²⁺ has been demonstrated to play an important role in cancer progression. Store-operated Ca²⁺ entry (SOCE) is the main Ca²⁺ entry pathway regulating intracellular Ca²⁺ concentration in a variety of cancer types. The present study aimed to explore the specific mechanisms of SOCE in the processes of glioma migration and invasion.

Methods

The expression of Orai1, a key component of SOCE, was examined in glioma samples and glioma cell lines by immunohistochemistry and western blot analysis. Both pharmacological intervention and RNA interference were employed to investigate the role of SOCE in glioma cell migration and invasion in vitro. The intracellular Ca²⁺ was certified through Fluo-4/AM based Ca²⁺ measurement. The effect of SOCE on cell viability, migration, and invasion was explored by methyl thiazolyl tetrazolium (MTT) assay, wound healing assay, transwell invasion assay. Western blot analysis and immunofluorescence assay were used to observe the changes of downstream related protein and cell morpholog.

Results

Orai1 expression was elevated in glioma tissues and several glioma cell lines compared with non-neoplastic brain tissues. Either inhibition of SOCE by a pharmacological inhibitor or Orai1 downregulation suppressed glioma cell migration and invasion. However, re-expression of Orai1 could rescue glioma cell motility. Furthermore, phosphorylation of proline-rich tyrosine kinase 2 (Pyk2) participated in the mechanisms by which SOCE regulated focal adhesion turnover and epithelial-to-mesenchymal (−like) transition in glioma cells, both of which are considered to be critical for tumor progression.

Conclusions

The SOCE-Pyk2 pathway is essential for glioma migration and invasion. The study indicates the potential value of Orai1 as a molecular target for anti-invasion therapy.

Electronic supplementary material

The online version of this article (doi:10.1186/s13046-014-0098-1) contains supplementary material, which is available to authorized users.     

Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其机制

目的： 探讨Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其作用机制。方法： 培养鼻咽上皮细胞NP69和4种鼻咽癌细胞系（CNE1、CNE2、HONE1和5-8F）,分别采用实时荧光定量PCR（qPCR）和Western blot法检测细胞中Orai1的mRNA和蛋白表达水平。采用靶向Orai1的短发夹RNA（shRNA）质粒,转染CNE2细胞,设转染对照质粒的CNE2细胞为阴性对照组,钙离子成像检测钙库调节的钙离子内流,Transwell小室检测细胞转移和侵袭,qPCR和Western blot法检测上皮-间充质转化（EMT）标志物E钙黏蛋白（E-cadherin）、N钙黏蛋白（N-cadherin）和波形蛋白（Vimentin）的mRNA和蛋白表达水平。结果： NP69、CNE1、CNE2、HONE1和5-8F细胞中均表达Orai1,且CNE2细胞中表达相对较高,故选择CNE2细胞进行后续试验。与阴性对照组相比,转染Orai1 shRNA后,CNE2细胞中Orai1的mRNA和蛋白水平均显著降低（P<0.05）;CNE2细胞的钙库调节钙离子内流能力、转移/侵袭能力和EMT相关标志蛋白E-cadherin、N-cadherin和Vimentin的mRNA和蛋白表达水平均显著下降（P<0.05）。结论： 抑制Orai1表达可抑制鼻咽癌细胞钙库调节钙离子内流、转移侵袭和EMT进程,提示Orai1可能成为治疗鼻咽癌转移的新靶标和新方向。     

Resveratrol activates autophagic cell death in prostate cancer cells via downregulation of STIM1 and the mTOR pathway

Resveratrol (RSV), a natural polyphenol, has been suggested to induce cell cycle arrest and activate apoptosis‐mediated cell death in several cancer cells, including prostate cancer. However, several molecular mechanisms have been proposed on its chemopreventive action, the precise mechanisms by which RSV exerts its anti‐proliferative effect in androgen‐independent prostate cancer cells remain questionable. In the present study, we show that RSV activates autophagic cell death in PC3 and DU145 cells, which was dependent on stromal interaction molecule 1 (STIM1) expression. RSV treatment decreases STIM1 expression in a time‐dependent manner and attenuates STIM1 association with TRPC1 and Orai1. Furthermore, RSV treatment also decreases ER calcium storage and store operated calcium entry (SOCE), which induces endoplasmic reticulum (ER) stress, thereby, activating AMPK and inhibiting the AKT/mTOR pathway. Similarly, inhibition of SOCE by SKF‐96365 decreases the survival and proliferation of PC3 and DU145 cells and inhibits AKT/mTOR pathway and induces autophagic cell death. Importantly, SOCE inhibition and subsequent autophagic cell death caused by RSV was reversed by STIM1 overexpression. STIM1 overexpression restored SOCE, prevents the loss of mTOR phosphorylation and decreased the expression of CHOP and LC3A in PC3 cells. Taken together, for the first time, our results revealed that RSV induces autophagy‐mediated cell death in PC3 and DU145 cells through regulation of SOCE mechanisms, including downregulating STIM1 expression and trigger ER stress by depleting ER calcium pool. © 2015 The Authors. Molecular Carcinogenesis, published by Wiley Periodicals, Inc.     

Store-operated Ca<Superscript>2+</Superscript> entry regulates glioma cell migration and invasion via modulation of Pyk2 phosphorylation

Background

The ubiquitous second messenger Ca²⁺ has been demonstrated to play an important role in cancer progression. Store-operated Ca²⁺ entry (SOCE) is the main Ca²⁺ entry pathway regulating intracellular Ca²⁺ concentration in a variety of cancer types. The present study aimed to explore the specific mechanisms of SOCE in the processes of glioma migration and invasion.

Methods

The expression of Orai1, a key component of SOCE, was examined in glioma samples and glioma cell lines by immunohistochemistry and western blot analysis. Both pharmacological intervention and RNA interference were employed to investigate the role of SOCE in glioma cell migration and invasion in vitro. The intracellular Ca²⁺ was certified through Fluo-4/AM based Ca²⁺ measurement. The effect of SOCE on cell viability, migration, and invasion was explored by methyl thiazolyl tetrazolium (MTT) assay, wound healing assay, transwell invasion assay. Western blot analysis and immunofluorescence assay were used to observe the changes of downstream related protein and cell morpholog.

Results

Orai1 expression was elevated in glioma tissues and several glioma cell lines compared with non-neoplastic brain tissues. Either inhibition of SOCE by a pharmacological inhibitor or Orai1 downregulation suppressed glioma cell migration and invasion. However, re-expression of Orai1 could rescue glioma cell motility. Furthermore, phosphorylation of proline-rich tyrosine kinase 2 (Pyk2) participated in the mechanisms by which SOCE regulated focal adhesion turnover and epithelial-to-mesenchymal (?like) transition in glioma cells, both of which are considered to be critical for tumor progression.

Conclusions

The SOCE-Pyk2 pathway is essential for glioma migration and invasion. The study indicates the potential value of Orai1 as a molecular target for anti-invasion therapy.

     

Inhibition of stromal‐interacting molecule 1‐mediated store‐operated Ca2+ entry as a novel strategy for the treatment of acquired imatinib‐resistant gastrointestinal stromal tumors

Imatinib has revolutionized the treatment of gastrointestinal stromal tumors (GIST); however, primary and secondary resistance to imatinib is still a major cause of treatment failure. Multiple mechanisms are involved in this progression. In the present study, we reported a novel mechanism for the acquired resistance to imatinib, which was induced by enhanced Ca²⁺ influx via stromal‐interacting molecule 1 (STIM1)‐mediated store‐operated Ca²⁺ entry (SOCE). We found that the STIM1 expression level was related to the acquired resistance to imatinib in our studied cohort. The function of STIM1 in imatinib‐resistant GIST cells was also confirmed both in vivo and in vitro. The results showed that STIM1 overexpression contributed to SOCE and drug response in imatinib‐sensitive GIST cells. Blockage of SOCE by STIM1 knockdown suppressed the proliferation of imatinib‐resistant GIST cell lines and xenografts. In addition, STIM1‐mediated SOCE exerted an antiapoptotic effect via the MEK/ERK pathway. The results from this study provide a basis for further research into potential novel therapeutic strategies in acquired imatinib‐resistant GIST.