CXCL12-CXCR4生物轴与卵巢癌关系的动物实验研究

目的探讨CXCL12。CXCR4趋化因子轴与卵巢癌发病的关系。方法将转染质粒SKOV3／CXCR4、转染载体SKOV3／neg及未转染的SKOV3三种卵巢癌细胞进行体外培养,建立荷人卵巢癌细胞裸鼠腹腔移植瘤模型,观察各组裸鼠自然死亡后腹腔成瘤数、体积、腹水量和存活期的变化。结果3组裸鼠生存时间、腹腔移植瘤的个数和重量比较差异有统计学意义（P〈0．05）,CXCL12浓度越高,受试裸鼠腹腔移植瘤个数越多,重量越大,生存时间越短。结论CXCL12-CXCR4趋化因子轴与卵巢癌的发生有一定的相关性,可能是其发病的一个重要原因,在一定程度上也反映了病情的轻重。     

子宫颈癌组织中 CXCL12-CXCR4生物轴及 MMP2-TIMP2的表达及意义

目的：探讨人子宫颈癌（宫颈癌）组织 CXCL12-CXCR4生物轴及 MMP2-TIMP2的表达,进一步研究宫颈癌发病的过程和转移机制。方法运用免疫组化法检测23例人宫颈癌患者（试验组）和因多发性子宫肌瘤行全子宫切除的患者25例（对照组）宫颈组织标本中 CXCL12-CXCR4和 MMP2-TIMP2的表达,研究其与子宫颈癌发病的相关性。结果与对照组相比宫颈癌组织中 CXCL12及 CXCR4阳性表达较多（ P <0.05）;与对照组相比,宫颈癌组织中 CXCL12及 CXCR4呈现高表达,二者与对照组比较,差异有统计学意义（ P <0.05）。与对照组相比宫颈癌组织中MMP-2及 TIMP-2阳性表达较多（ P <0.05）,与对照组相比,宫颈癌组织中 MMP-2及 TIMP-2呈现高表达（ P <0.05）。CXCL12与 MMP2呈正相关（ r =0.452,P <0.05）;CXCL12与 TIMP2呈正相关（ r =0.533,P <0.05）。结论宫颈癌发病和转移可能与 CXCL12-CXCR4生物轴及 MMP2-TIMP2密切相关,CXCL12是在其中起着重要作用。     

CXCL12-CXCR4生物轴与卵巢癌关系的动物实验研究

目的：探讨CXCL12-CXCR4趋化因子轴与卵巢癌发病的关系。方法将36只裸鼠按照体重大小编号,随机分为3组,将转染质粒SKOV3／CXCR4、转染载体SKOV3／neg及未转染的SKOV3三种卵巢癌细胞进行体外培养,取三种细胞悬液4×106个（200μl）于裸鼠左下腹注入腹腔,建立荷人卵巢癌细胞裸鼠腹腔移植瘤模型。自注入肿瘤细胞后第2天起,将每组裸鼠又随机分为对照组和处理组,对照组腹腔内注射0い．9％氯化钠溶液0．2 ml,处理组腹腔内注射CXCL12-CXCR4拮抗剂AMD3100,观察各组裸鼠自然死亡后腹腔成瘤数、体积、腹水量和存活期的变化。结果 SKOV3／CXCR4、SKOV3／neg及SKOV3三种卵巢癌细胞接种于裸鼠腹腔后1周左右荷瘤裸鼠在对照组和处理组体重差比较,差异有统计学意义（ P ＜0．05）。接种SKOV3／CXCR4细胞的裸鼠对照组的平均生存期与CXCR4拮抗剂AMD3100处理组裸鼠比较,差异有统计学意义（ Z ＝－2．29, P ＜0．05）;而接种SKOV3／neg细胞的裸鼠对照组的平均生存期与CXCR4拮抗剂AMD3100处理组裸鼠比较,差异无统计学意义（ P ＞0．05）;接种SKOV3细胞的裸鼠平均生存期与CXCR4拮抗剂AMD3100处理的裸鼠比较,差异无统计学意义（ P ＞0．05）。接种SKOV 3／CXCR4细胞裸鼠的对照组腹腔移植瘤的平均个数和平均重量与CXCR4拮抗剂AMD3100处理组比较,差异有统计学意义（ P ＜0．05）。结论 CXCL12-CXCR4趋化因子轴与卵巢癌的发生有一定的相关性,可能是其发病的一个重要原因,在一定程度上也反映了病情的轻重。     

苯并[a]芘通过CXCL12/CXCR4轴影响早孕小鼠子宫内膜炎症反应CSCD

目的探讨CXCL12/CXCR4轴在苯并[a]芘(Benzo[a]pyrene,B[a]P)影响早孕小鼠胚胎着床过程中的作用及其机制。方法将8周龄昆明小鼠每晚按雌雄2∶1比例合笼,次晨将查得阴栓者记为孕第1天(D1),并随机分为对照组和B[a]P组。B[a]P组孕鼠每日晨称重后以0.1 ml/10 g动物体重灌胃给予0.2 mg/(kg·d)B[a]P,对照组则灌胃等体积玉米油。体外分离小鼠原代子宫内膜基质细胞,将其分为对照组、B[a]P组和rhCXCL12+BaP联合处理组。ELISA法检测小鼠血清CXCL12水平;qRT-PCR、Western blot和免疫组化法检测CXCL12、CXCR4的表达;qRT-PCR和Western blot检测炎症因子IL-1β、IL-10、NLRP3和TNF-α的表达;Western blot检测着床相关因子MUC1、MMP9和HOXa10的表达情况。结果与对照组相比,B[a]P暴露明显降低孕早期小鼠血清中CXCL12含量,同时下调小鼠子宫组织及基质细胞内CXCL12和CXCR4的表达,抑制炎症反应的发生,影响着床相关因子表达。上调小鼠原代子宫内膜基质细胞内CXCL12表达,可缓解B[a]P所导致的炎症抑制和着床相关因子表达异常。结论B[a]P可能通过下调CXCL12/CXCR4表达,抑制炎症反应,影响孕早期小鼠胚胎植入过程。     

CXCR4抑制剂与血液系统肿瘤的治疗

基质细胞衍生因子(SDF-1)又名CXCL12或前B细胞刺激因子(pre-B cell stimulatory factor,PBSF),是主要由骨髓基质细胞和不成熟的成骨细胞产生的趋化因子。SDF-1表达于骨髓、淋巴结、肺、大脑、心脏、肝脏、胸腺、脾脏和肾脏等组织和器官。SDF-1受体之一的CXCR4是一个具有7个跨膜受体的G蛋白偶联受体,定位于第4号染色体,具有类似于白介素(IL)-8受体的结构,表达于造血干/祖细胞、血液和骨髓的多种细胞。     

卵巢肿瘤干细胞中趋化因子的表达与细胞侵袭转移能力的关系

目的探讨卵巢肿瘤干细胞中趋化因子的表达与细胞侵袭转移能力的关系,为卵巢肿瘤的治疗提供新的研究思路和药物作用靶点。方法采用流式细胞术与RT-PCR方法检测卵巢肿瘤干细胞系CAOV-3中CXCR4的表达,通过体外微孔隔离室（Transwell）检测CXCR4对CAOV-3细胞侵袭转移能力的影响。结果 CXCR4在CAOV-3中细胞呈阳性表达,CXCL12可促进CAOV-3细胞的迁移,CXCR4的封闭能抑制CXCL12对CAOV-3迁移的促进作用。结论 CXCL12-CXCR4相互作用可促进卵巢肿瘤干细胞侵袭转移,在卵巢肿瘤的生长和侵袭转移过程中起着重要作用。     

小窝蛋白-1与特发性肺纤维化

小窝蛋白作为小窝膜内的支架蛋白,能够组织和浓缩特异性的脂质,修饰信号转导分子以及负性调控许多信号转导通路和生物过程。生物体内小窝蛋白表达异常可引起疾病,特发性肺纤维化就是其中一种。大量研究表明,小窝蛋白-1在特发性肺纤维化发病过程中表达明显下调,大大降低了其对多条信号传导通路如TGF-β/SMAD、ERK/MAPK、JNK/MAPK和PKC等的负性调控作用,进而导致肺泡上皮细胞损伤、成纤维细胞增殖及表型转化、细胞外基质(ECM)重塑,加速了肺纤维化发病进程。因此该文就小窝蛋白-1在特发性肺纤维化中的作用进行了综述。     

影响CXCL12/CXCR4在NSCLC中表达的相关性研究

目的 检测CXCL12、CXCR4在非小细胞肺癌(NSCLC)组织及癌旁正常肺组织中的表达情况,分析其阳性表达的意义及其阳性表达与NSCLC相关参数的联系。方法 采用免疫组织化学染色方法对临床确诊的NSCLC癌组织、离其最短径5 cm以上正常肺组织标本62例进行检测。结果 NSCLC细胞CXCL12、CXCR4与NSCLC癌旁正常肺组织阳性表达率差异有统计学意义(P<0.05);与NSCLC的TNM分期、纵隔淋巴结转移等差异有统计学意义(P<0.05)。结论 CXCL12、CXCR4在NSCLC组织中表达能够促进NSCLC癌组织的浸润、发展及转移,并且与纵隔淋巴结转移有关;其表达是专一性的,具有靶向性。     

趋化因子CXCL12/CXCR4在非小细胞肺癌中的表达

目的:探讨趋化因子CXCL12及其受体CXCR4在非小细胞肺癌(NSCLC)中的表达及与临床病理特征间的关系。方法:采用免疫组织化学SP三步法检测95例NSCLC患者肿瘤组织及27例肺部良性肿瘤患者(对照组)的石蜡标本中CXCL12、CXCR4的表达情况,并比较其在不同临床病理参数患者间表达的差异。结果:对照组中,CXCL12和CXCR4的高表达率分别为0和7.4%(2/27),NSCLC患者的高表达率分别为54.7%(52/95)和42.1%(40/95),差异均有统计学意义(P<0.01)。在NSCLC中,淋巴结转移组的CXCL12和CXCR4的高表达率(67.2%、51.7%)均高于非转移组(35.1%、27.0%),差异均有统计学意义(P<0.05)。CXCL12、CXCR4的高表达率均随TNM分期的增高而逐渐增高,且Ⅲ期的高表达率明显高于Ⅰ期,差异有统计学意义(P<0.01)。NSCLC患者CXCL12与CXCR4之间无明显相关性。不同年龄、性别、肿瘤大小、组织学分型及分化程度NSCLC患者之间CXCL12、CXCR4的高表达率差异无统计学意义(P>0.05)。结论:CXCL12、CXCR4的表达水平可作为判断NSCLC预后的参考指标。     

趋化因子受体CXCR4及其配体CXCL12在肝癌侵袭 转移中的作用及机制

目的 探讨趋化因子受体 CXCR4 及其配体 CXCL12（CXCL12/CXCR4）在原发性肝癌侵袭转移中的作用 及机制。方法 分别采用 Western blot、免疫组化和 Real-time PCR 等方法检测 60 例肝癌及对应癌旁组织标本中 CXCL12/CXCR4 蛋白及 mRNA 表达水平。常规培养 4 种肝癌细胞（Huh7、MHCC97h、HepG2、Hep3B）和正常肝细胞 （7702）,Real-time PCR 检测上述细胞中 CXCL12、CXCR4 mRNA 的表达,筛选合适的实验细胞。将 CXCR4 干扰质 粒（sh-CXCR4）和对应空载体（sh-control）分别转染至 MHCC97h 构建稳转细胞系。Transwell 侵袭实验、细胞划痕实 验、MTT 实验分别检测 2 组细胞的侵袭、迁移和增殖能力。取稳定表达的 sh-control 和 sh-CXCR4 MHCC97h 细胞, 接种至 6 只裸鼠皮下,观察瘤体生长情况。Western blot 检测 sh-control 和 sh-CXCR4 MHCC97h 细胞及对应裸鼠移 植瘤中血管内皮生长因子-C（VEGF-C）的表达,同时对转染 CXCR 过表达质粒的 MHCC97h 中 VEGF-C 的表达水平 进行检测。结果 （1）Western blot、免疫组化和 Real-time PCR 的结果均证实肝癌组织中 CXCL12/CXCR4 蛋白及 mRNA 表达水平均高于癌旁组织。（2）Huh7、MHCC97h、HepG2、Hep3B 细胞中 CXCL12/CXCR4 mRNA 表达水平均高 于 7702 细胞,选取 MHCC97h 为实验细胞。转染 sh-CXCR4 的 MHCC97h 细胞的侵袭、迁移和增殖能力均明显低于 sh-control 组,同时接种含 sh-CXCR4 的 MHCC97h 细胞的裸鼠移植瘤的生长速度也明显小于 sh-control 组。（3）体外 和体内实验均证实 sh-CXCR4 组的 VEGF-C 表达水平均低于 sh-control 组,而过表达 CXCR4 后,VEGF-C 的蛋白表 达明显上调。结论 CXCL12/CXCR4 在原发性癌组织和肝癌细胞中呈现高表达,CXCL12/CXCR4 可能通过调控 VEGF-C 蛋白表达从而抑制肝癌细胞的增殖、侵袭和转移。     

CXCL16/CXCR6 axis promotes bleomycin-induced fibrotic process in MRC-5 cells via the PI3K/AKT/FOXO3a pathway

ObjectiveInterstitial lung disease (ILD) is a progressive and irreversible lung disease with very limited therapeutic options. Previous studies have found that chemokine ligands CXCL16 and CXCR6 play critical roles in organ fibrosis. However, whether CXCL16 and CXCR6 are also involved in the pathogenesis of ILD, as well as their regulatory role in pulmonary fibrosis, has not been reported.MethodsIn this study, we detected CXCL16 levels in patients with rheumatoid arthritis-associated ILD (RA-ILD) and examined the critical role of the CXCL16/CXCR6 axis in the proliferation and collagen production of human pulmonary fibroblasts (MRC-5 cells). The effect of anti-CXCL16 antibody on the bleomycin-induced fibrogenesis in cultured MRC-5 cells was also evaluated.ResultsOur results indicated that serum soluble CXCL16 was significantly higher in RA-ILD patients and also associated with the severity of lung fibrosis. CXCL16 facilitates fibrosis by enhancing proliferation, migration, and collagen production of MRC-5 cells. Furthermore, a synergistic fibrogenic effect of CXCL16 and bleomycin has been found. CXCL16 stimulated the activation of PI3K/AKT/FOXO3a signaling pathway in MRC-5 cells, and the inhibition by specific inhibitors Wortmannin and LY294002, or knockdown of CXCR6 by siRNA also suppressed the biological functions of MRC-5 cells mediated by CXCL16. Similarly, down-regulation of CXCR6 also partly blocked BLM-induced fibrogenesis in MRC-5 cells.ConclusionsCXCL16/CXCR6 axis promotes proliferation and collagen production of MRC-5 cells by the PI3K/AKT/FOXO3a signaling pathway, and inhibition of the CXCL16/CXCR6 axis may provide a new therapeutic strategy targeting pulmonary fibrosis.     

CXCR4 chemokine receptor antagonists: perspectives in SCLC

Small-cell lung cancer (SCLC) is a particularly aggressive form of lung cancer characterized by early and widespread metastases and the ability to rapidly develop resistance against chemotherapeutic agents. Tumor cell migration and metastasis share many similarities with leukocyte trafficking, which is critically regulated by chemokine receptors and adhesion molecules. SCLC cells express high levels of CXCR4 (CD184), a seven-transmembrane G-protein-coupled chemokine receptor. Stromal cells within the bone marrow microenvironment and at extramedullary sites constitutively secrete stromal cell-derived factor-1 (CXCL12), the ligand for CXCR4. Activation of CXCR4 induces SCLC cell migration and adhesion to stromal cells that secrete CXCL12, which in turn provides growth- and drug resistance-signals to the tumor cells. CXCR4 antagonists, such as Plerixafor (AMD3100) and T140 analogues (TN14003/ BKT140), disrupt CXCR4-mediated SCLC cell-adhesion to stromal cells. In stromal cell co-cultures, CXCR4 antagonists also sensitize SCLC cells to cytotoxic drugs, such as etoposide, and thereby antagonize cell adhesion-mediated drug resistance. Therefore, targeting the CXCR4–CXCL12 axis is a novel, attractive therapeutic approach in SCLC. Here, we summarize preclinical data about CXCR4 in SCLC, and the current status of the preclinical and clinical development of CXCR4 antagonists.     

A proposed mechanism for the protective effect of dioxin against breast cancer.

Although it is causative for many types of cancers, experimental and epidemiological evidence suggest that 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) may in fact protect against breast cancer. The mechanism(s) for this protection remain unclear. In an attempt to further elucidate this mechanism, we performed a microarray experiment to identify genes that were modulated upon dioxin treatment. We found that dioxin downregulated the messenger RNAs for the G-protein-coupled receptor, CXCR4, as well as its unique chemokine ligand, CXCL12, in MCF-7 breast cancer cells. We demonstrated that the corresponding proteins are also downregulated by dioxin. The interaction between CXCR4 and CXCL12 plays a central role in the metastasis of breast cancer, as disruption of the CXCL12/CXCR4 axis has been shown to limit the metastasis of breast cancer cells to the lung in mice. Utilizing an in vitro chemotaxis assay, we demonstrate that dioxin specifically inhibits the migration of MCF-7 cells toward CXCL12. We also show that dioxin reduces CXCR4 under hypoxia and CXCL12 under estradiol-induced conditions in MCF-7 cells. Finally, as the CXCR4/CXCL12 axis is implicated in the progression of numerous types of cancer, we identified several other cancer cell lines in which dioxin modulates CXCR4 and CXCL12 levels. We therefore propose that one mechanism whereby dioxin may protect against breast cancer is via downregulation of CXCR4 and CXCL12, thereby inhibiting progression of the disease. Further, other nontoxic ligands for the aryl hydrocarbon receptor (selective AHR modulators) may exert their protective effects by a similar mechanism.     

Inhibition of CXCL12/CXCR4 autocrine/paracrine loop reduces viability of human glioblastoma stem-like cells affecting self-renewal activity

Cancer stem cells (CSCs) or tumor initiating cells (TICs) drive glioblastoma (GBM) development, invasiveness and drug resistance. Distinct molecular pathways might regulate CSC biology as compared to cells in the bulk tumor mass, representing potential therapeutic targets. Chemokine CXCL12 and its receptor CXCR4 control proliferation, invasion and angiogenesis in GBM cell lines and primary cultures, but little is known about their activity in GBM CSCs. We demonstrate that CSCs, isolated from five human GBMs, express CXCR4 and release CXCL12 in vitro, although different levels of expression and secretion were observed in individual cultures, as expected for the heterogeneity of GBMs.CXCL12 treatment induced Akt-mediated significant pro-survival and self-renewal activities, while proliferation was induced at low extent. The role of CXCR4 signaling in CSC survival and self-renewal was further demonstrated using the CXCR4 antagonist AMD3100 that reduced self-renewal and survival with greater efficacy in the cultures that released higher CXCL12 amounts. The specificity of CXCL12 in sustaining CSC survival was demonstrated by the lack of AMD3100-dependent inhibition of viability in differentiated cells derived from the same GBMs.These findings, although performed on a limited number of tumor samples, suggest that the CXCL12/CXCR4 interaction mediates survival and self-renewal in GBM CSCs with high selectivity, thus emerging as a candidate system responsible for maintenance of cancer progenitors, and providing survival benefits to the tumor.     

Oligomannurarate sulfate inhibits CXCL12/SDF-1-mediated proliferation and invasion of human tumor cells in vitro

Aim:

JG6 is a novel marine-derived oligosaccharide that has shown to inhibit angiogenesis and tumor metastasis. In this study, we sought to identify the potential target responsible for the anti-cancer activity of JG6.

Methods:

Human liver cancer cell line Bel-7402 and human cervical cancer cell line HeLa were examined. CXCL12-stimulated cell proliferation and migration were determined using a CCK-8 kit and a transwell assay, respectively. Western blotting was performed to examine the changes in CXCL12/CXCR4 axis. Molecular docking and surface plasmon resonance (SPR) were performed to characterize the possible interaction between JG6 and the CXCL12/CXCR4 axis.

Results:

Treatment with CXCL12 potently stimulated the proliferation and migration in both Bel-7402 and HeLa cells. Co-treatment of the cells with JG6 (10, 50 and 100 μg/mL) dose-dependently impeded the CXCL12-stimulated cell proliferation and migration. Furthermore, CXCL12 rapidly induced phosphorylation of AKT, ERK, FAK and Paxillin in Bel-7402 and HeLa cells, whereas pretreatment with JG6 dose-dependently inhibited the CXCL12-induced phosphorylation of these proteins. The SPR assay showed that JG6 bound to CXCL12 with a high affinity. In molecular docking study, JG6 appeared to interact with CXCL12 via multiple polar interactions, including 6 ionic bonds and 7 hydrogen bonds.

Conclusion:

Inhibition of the CXCL12/CXCR4 axis by JG6 may account for its anticancer activity.     

乳腺癌肿瘤细胞中趋化因子CXCL12及其受体CXCR4的研究进展

CXCL12因子及其受体CXCR4作为趋化因子家族中与肿瘤关系最为密切的一对。特别在乳腺癌方面,无论是对原位肿瘤还是转移性肿瘤都进行了一系列深入的研究,本文就这一方面的研究进行了总结和展望。     

Development of novel CXCR4-based therapeutics

Introduction: During embryogenesis, CXCR4, a chemokine receptor, and its ligand, stromal cell-derived factor-1 (SDF-1/CXCL12), are critically involved in the development of the hematopoietic, nerve and endothelial tissues by regulating tissue progenitor cell migration, homing and survival. In adult life, the CXCR4 axis serves as the key factor for stem and immune cell trafficking. More importantly, CXCR4–CXCL12 axis plays a critical role in HIV, stem cell mobilization, autoimmune diseases, cancer and tissue regeneration. Targeting the CXCR4–CXCL12 axis, therefore, is an attractive therapeutic approach in various diseases.

Areas covered: In this review, we update current knowledge about CXCR4–CXCL12 biology, therapeutic approaches and therapeutic agents. The data presented was collected from http://www.ncbi.nlm.nih.gov/pubmed, http://clinicaltrials.gov/, http://bloodjournal.hematologylibrary.org/.

Expert opinion: Development of CXCR4 antagonists with increased affinity, extended pharmacokinetics and/or pharmacodynamics and with the capacity to differentially target CXCR4 may lead to a development of novel therapeutics for HIV, cancer, tissue regeneration and stem cell collection.     

CXCL12/CXCR4在肾癌中的研究进展

肾癌是泌尿系统常见的恶性肿瘤之一,早期症状不明显,发现时往往已发生了转移。近年来发现趋化因子CXCL12及其受体CXCR4在肾癌转移中发挥着重要的作用,对CXCR4检测有利于判断肾癌转移及患者的预后。CXCR4靶向治疗已在动物模型及临床前试验中取得了成功,有望成为治疗肾癌新的突破口。