TPX2与肿瘤关系的研究进展

基因组不稳定是癌症的标志.XKlP2靶蛋白(Targeting protein for Xenopus kinesin-like protein 2,TPX2)在细胞有丝分裂过程中参与纺锤体的形成.TPX2的异常表达可导致中心体异常扩增、纺锤体发生异常,进而影响染色体组的稳定性,细胞发生恶性转化.近年来研究证实,TPX...     

下调TPX2抑制喉癌细胞生长的体外实验研究

目的:研究TPX2对喉癌细胞增殖、克隆形成能力及细胞周期的影响。方法:Hep-2细胞中转染TPX2 siRNA、siRNA control记为TPX2 siRNA、siRNA -NC组，以不做转染的细胞为Con组。荧光定量PCR和Western blot分别测定细胞中TPX2 mRNA和蛋白水平，MTT测定各组细胞增殖，平板克隆实验测定各组细胞克隆形成能力，流式细胞术测定各组细胞周期情况，Western blot测定各组细胞中增殖细胞核抗原（PCNA）、细胞核增殖抗原(Ki-67)、细胞周期依赖性蛋白激酶4(CDK4)、细胞周期蛋白D1（Cyclin D1）蛋白水平。结果:siRNA control中TPX2 mRNA和蛋白水平、细胞存活率、克隆形成数目、细胞周期以及细胞中PCNA、Ki-67、CDK4、Cyclin D1蛋白水平与Con相比均没有明显变化（P＞0.05）。TPX2 siRNA细胞中TPX2 mRNA和蛋白水平均明显低于Con（P＜0.05）。TPX2 siRNA细胞存活率、克隆形成数目均明显低于Con，细胞G0/G1期比例明显高于Con，细胞中PCNA、Ki-67、CDK4、Cyclin D1蛋白水平明显低于Con（P＜0.05）。结论:TPX2敲低可以降低喉癌细胞增殖、克隆形成能力，将细胞周期阻滞在G0/G1期，降低细胞中PCNA、Ki-67、CDK4、Cyclin D1蛋白表达。     

短发夹RNA靶向沉默TPX2基因促进人肺腺癌A549细胞凋亡及其相关机制

目的:探讨靶向Xklp2靶蛋白(targeting protein for Xenopus kinesin-like protein2,TPX2)基因的短发夹RNA(short hairpin RNA,shRNA)对肺腺癌A549细胞凋亡的影响及其可能机制。方法:构建靶向TPX2基因的shRNA重组载体,将其转染至肺腺癌A549细胞中,RT-PCR检测细胞中TPX2、Aurora-A、p53和Bcl-2mRNA的表达,蛋白质印迹法检测TPX2蛋白的表达,FCM检测细胞周期和细胞凋亡情况。结果:成功构建重组载体pMagic4.1-shRNA-TPX2。将pMagic4.1-shRNA-TPX2转染至A549细胞后,TPX2、Aurora-A和Bcl-2mRNA的表达水平明显下调,p53mRNA的表达水平明显上调,TPX2蛋白的表达水平明显下调,细胞凋亡率明显增加,细胞阻滞于S期,与空白对照组(未转染组)和阴性对照组(转染pMagic4.1-shRNA-NC)相比,差异均有统计学意义(P<0.05)。结论:靶向TPX2的shRNA能促进肺腺癌A549细胞的凋亡,其作用可能与上调p53表达和下调Bcl-2表达有关。     

TPX2在非肌层浸润性膀胱癌中的表达及其临床意义

目的： 研究Xklp2靶蛋白（TPX2）在非肌层浸润性膀胱癌中的表达及其临床意义。方法： 通过挖掘GEPIA数据库中的研究数据,对TPX2 mRNA在膀胱癌组织中的表达及其与患者生存的关系进行分析;采用免疫组织化学染色检测TPX2蛋白在60例非肌层浸润性膀胱癌石蜡标本组织中的表达,并分析其表达与肿瘤临床病理指标和患者预后的关系。结果： GEPIA数据库分析提示TPX2 mRNA在膀胱癌组织中的表达较正常组织升高（P<0.05）,且TPX2 mRNA高表达组患者的无病生存率较低表达组患者降低（P<0.05）;免疫组织化学染色检测结果显示TPX2蛋白在非肌层浸润性膀胱癌组织中的阳性表达率为75%（45/60）,而在对应的基底组织中,TPX2蛋白表达均为阴性,二者之间的差异具有统计学意义（P<0.01）;且TPX2的表达水平与患者肿瘤数目、肿瘤直径、病理分级及术后复发等病理指标有关（P<0.05）,生存分析提示TPX2高表达的膀胱癌患者的生存期较低表达者短（P<0.05）。结论： TPX2在非肌层浸润性膀胱癌中表达上调,且其表达水平较高的患者预后较差,TPX2可能作为预测非肌层浸润性膀胱癌患者预后的肿瘤标记物。     

Aurora激酶与人类肿瘤的研究进展

Aurora蛋白激酶家族,包括Au-rora-A、Aurora-B和Aurora-C,在细胞有丝分裂期过程中发挥着重要作用。Aurora-A定位于中心体和纺锤体,主要参与中心体的成熟、分离和纺锤体形成,还参与p53通路、细胞凋亡和有丝分裂的调节等。Auro-ra-A在多种肿瘤组织中存在着高表达。它的高表达可能会导致基因的不稳定和肿瘤的发生,与某些肿瘤的预后相关。最近对其三维结构的测定推进了对激酶抑制剂的研究,希望能够发现新的抗癌因子。Aurora-B参与染色体调节、分离,并在纺锤体检查点等方面发挥重要作用。Auro-ra-C和Aurora-B共同参与哺乳动物有丝分裂染色体分离和胞质分裂。     

TPX2在肾透明细胞癌中的表达及其临床意义

目的分析Xklp2靶蛋白(TPX2)在肾透明细胞癌(KIRC)组织中的表达及其临床意义。方法收集2017年7月至2019年6月在蚌埠医学院第一附属医院泌尿外科收治的54例KIRC患者术后组织标本。利用免疫组织化学法检测TPX2在KIRC组织和癌旁组织的蛋白表达。利用TIMER数据库分析TPX2 mRNA在KIRC组织与正常组织的表达差异, 验证免疫组织化学结果。使用UALCAN数据库和Kaplan-Meier plotter数据库对TPX2 mRNA表达与KIRC患者临床分期、分子亚型、淋巴结转移及预后的关系进行分析。通过STRING数据库进行蛋白互作网络构建获得TPX2相关蛋白, 将相关蛋白对应的基因进行KEGG通路富集。利用TIMER数据库对TPX2的表达与免疫细胞浸润、免疫检查点的关系进行研究。结果免疫组织化学结果显示, 癌组织中的TPX2蛋白的阳性表达率(48.15%, 26/54)高于癌旁组织中阳性表达率(20.37%, 11/54)(χ2=9.25, P=0.002)。生物信息学分析结果显示, TPX2 mRNA表达水平在KIRC中显著上调[癌组织:1.89(1.49, 2...     

miR-218-5p靶向TPX2调节p53通路影响肺腺癌恶性进展

背景与目的肺腺癌(lung adenocarcinoma,LUAD)是肺癌的一种主要亚型,其治疗与诊断依然是目前的研究热点。靶向Xklp2靶蛋白(targeting protein for Xenopus kinesin-like protein2,TPX2)在多种癌细胞中高表达,可能与LUAD的发生发展相关。本研究旨在探究TPX2对LUAD细胞恶性进程的影响以及调控机制。方法通过生物信息学分析技术,对癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中LUAD组织中基因TPX2的表达情况进行分析。实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测人肺正常细胞系和人LUAD细胞系中TPX2和miR-21 8-5p的表达水平。蛋白质印迹法(Western blot)检测细胞系中TPX2蛋白表达以及其对p53信号通路关键蛋白表达的影响。使用生物信息学预测并通过双荧光素酶报告基因检测验证TPX2与miR-218-5p的关系,细胞活力检测(cell counting k...     

细胞周期蛋白cyclin B1与肿瘤

细胞周期蛋白cyclin B1与细胞周期蛋白依赖性蛋白激酶（cyclin-dependent protein kinases,CDKs）结合,受磷酸化、去磷酸化调节,所形成的活性复合物为MPF即细胞促分裂因子或M期促进因子,能转位入核,磷酸化其核内底物,促进细胞的G2/M期转变;并能通过磷酸化调节多种蛋白的活性与分布而参与纺锤体的形成和染色体的分离。cyclin B1在许多肿瘤中异常表达,呈现出癌基因的特性,cyclin B1过表达与定位的改变受p53、c-myc、H-Ras、Aurora A、Gadd45等多个基因的调节,而且与肿瘤发生发展、预后、诊断和治疗密切相关。     

癌基因B-RafV600E导致黑色素瘤Sbcl2和SK-MEL31细胞染色体不稳定性的分子机制研究

目的 研究癌基因B-RafV600E导致肿瘤细胞染色体不稳定的分子机制.方法 采用RNA干扰技术敲除稳定表达B-RafV600E基因的黑色素瘤Sbcl2和SK-MEL31细胞中内源性单核纺锤体蛋白激酶(Mps1)基因表达,免疫荧光染色技术检测中心体及纺锤体结构.HU-arrest分析法观察Mps1基因缺失对癌基因B-RafV600E致肿瘤细胞中心体过度复制及多极纺锤体形成的影响.结果 未敲除内源性Mps1基因的表达B-RafV600E基因的Sbcl2和SK-MEL31细胞中36％出现中心体过度复制及多极纺锤体,Mps1基因被敲除后上述异常细胞比例降低至6％.结论 B-RafV600E可能通过Mps1调控中心体过度复制及多极纺锤体结构的形成,影响肿瘤细胞染色体不稳定性及非整倍体细胞的出现.     

肺癌患者血清TPX2、GASP-1表达水平及其与病理分型的相关性分析

目的 研究血清Xklp2靶蛋白(TPX2)、G蛋白偶联受体相关分选蛋白1(GASP-1)水平与肺癌病理特征的关系。方法 将160例肺癌患者纳为研究对象,均为首次确诊,并未接受过任何肺癌相关治疗,采集其外周静脉血,检测血清TPX2及GASP-1水平,分析不同病理类型肺癌患者血清TPX2及GASP-1水平的差异。结果 与健康对照组比较,肺癌组患者血清TPX2及GASP-1水平均显著升高(P<0.05);血清TPX2及GASP-1对肺癌有良好的诊断效能,而两者联合应用诊断效能更高。小细胞肺癌(SCLC)患者血清TPX2及GASP-1水平显著高于非小细胞肺癌(NSCLC)患者(P<0.05),此外,肺腺癌患者血清TPX2及GASP-1水平显著高于肺鳞癌患者(P<0.05)。Ⅰ期及Ⅱ期肺癌患者血清TPX2及GASP-1无显著性差异(P>0.05),Ⅲ期及Ⅳ期肺癌患者血清TPX2及GASP-1水平均显著高于Ⅰ期及Ⅱ期者(P<0.05),且Ⅳ期患者血清TPX2及GASP-1水平显著高于Ⅲ期者(P<0.05)。低分化肺癌患者血清TPX2及GASP-1水平显著高于中、...     

TPX2 Overexpression in Medullary Thyroid Carcinoma Mediates TT Cell Proliferation

TPX2 (targeting protein for xenopus kinesin-like protein 2), a microtubule-associated protein, plays an important role in the formation of the mitotic spindle. Abnormal expression of TPX2 in various types of malignant tumors has been reported, but less is known for medullary thyroid cancer (MTC). We investigated the expression of TPX2 in human MTC tissues and its potential use as a therapeutic target. Immunohistochemical analysis of TPX2 expression was performed for 32 cases of MTC and 8 cases of normal thyroid. TPX2 expression was found to be significantly higher in MTC compared to normal thyroid tissues (P?<?0.05), and to be associated with tumor size, lymph node metastasis, and advanced disease stage. The cellular effects of TPX2 knockdown, including cell proliferation, apoptosis, cell cycle diffusions, and mitotic gene expression were investigated using small interfering RNA (siRNA). TPX2-siRNA caused G1 and G2-phase cell cycle arrest, inhibited cell proliferation, and induced apoptosis. TPX2-siRNA also downregulated Aurora-A and cyclinB1 protein expression in MTC cells and enhanced the expression of p53 protein (P?<?0.05). These results suggest that TPX2 may be of potential use as a new marker for MTC prognosis and therapy.     

The TPX2 gene is a promising diagnostic and therapeutic target for cervical cancer

The target protein for Xklp2 (TPX2), a microtubule-associated protein, can be used to evaluate more precisely the proliferative behavior of tumor cells. The abnormal expression of TPX2 in various types of malignant tumors has been reported, but less is known for cervical cancer. We studied the relationship between TPX2 expression and the biological behavior of cervical cancer. Immunohistochemistry and RT-PCR were used to detect the expression of TPX2 in cervical cancer tissues. The inhibitory effect of TPX2-siRNA on the growth of HeLa human cervical carcinoma cells was studied in?vitro. TPX2 expression was found to be significantly higher in cervical carcinoma compared to normal cervical tissues and CIN. The expression of TPX2 in cervical cancer was correlated with histological grading, FIGO staging and lymph node metastasis. TPX2 RNAi in HeLa cervical cancer cells caused S-phase cell cycle arrest, induced apoptosis and inhibited cell proliferation and invasion. In conclusion, TPX2 shows potential to be used as a new marker for cervical cancer diagnosis and therapy.     

Overexpressed targeting protein for Xklp2 (TPX2) serves as a promising prognostic marker and therapeutic target for gastric cancer

The targeting protein for Xenopus kinesin-like protein 2 (TPX2) is a putative oncogene in different human cancers. This study assessed TPX2 expression in gastric cancer tissue samples and then determined the effects of TPX2 knockdown on the regulation of gastric cancer cell malignant behaviors in vitro. Tissue samples from 115 gastric cancer patients were analyzed for TPX2 expression. The effects of TPX2 siRNA on gastric cancer cells were assessed in vitro, including cell viability, cell cycle distribution, apoptosis, migration, and invasion. The data showed that TPX2 was overexpressed in gastric cancer tissues compared to that in the adjacent normal epithelia. Moreover, TPX2 overexpression was associated with a poor overall survival and was an independent prognostic predictor of gastric cancer. In addition, the in vitro study further confirmed the ex vivo data, i.e., knockdown of TPX2 expression reduced gastric cancer cell viability but induced apoptosis and arrested cells at the G2/M phase of the cell cycle. Knockdown of TPX2 expression also inhibited the tumor cell migration and invasion capacity in vitro. At the gene level, knockdown of TPX2 expression upregulated the levels of cyclin B1, cdk4, p53, Bax, caspase-3, and E-cadherin, but downregulated the levels of cyclin D1, cdk2, N-cadherin, slug, matrix metalloprotease (MMP)-2, and MMP-9, suggesting that knockdown of TPX2 expression suppressed tumor cell epithelial–mesenchymal transition (EMT). This study demonstrated that detection of TPX2 overexpression could serve as a prognostic marker and therapeutic target for gastric cancer.     

Receptor for hyaluronan-mediated motility correlates with centrosome abnormalities in multiple myeloma and maintains mitotic integrity

Elevated expression of receptor for hyaluronan-mediated motility (RHAMM) within ex vivo diagnostic multiple myeloma plasma cells predicts for aggressive disease and patient survival. Here, we investigate the relationship between RHAMM and centrosomal abnormalities within multiple myeloma patient samples. We report that myeloma patient samples contain pervasive structural and numerical centrosomal abnormalities. Structural, but not numerical, centrosomal abnormalities strongly correlate with elevated RHAMM expression. As others have shown that excess pericentriolar material strongly associates with abnormal mitoses, we modeled centrosomal abnormalities with exogenous RHAMM overexpression. RHAMM overexpression in vitro resulted in centrosomal and mitotic defects. To elucidate a mechanism for RHAMM-mediated spindle defects, we further investigated RHAMM mitotic function. RHAMM mitotic localization mirrors that of targeting protein for Xklp2 (TPX2), and RHAMM interacts with the spindle assembly factors dynein and TPX2. Like TPX2, RHAMM expression is up-regulated during mitosis. Moreover, inhibition of function experiments reveals that RHAMM and TPX2 functions converge to maintain spindle integrity after spindle assembly. We postulate that augmentation of RHAMM expression within human cancers, including myeloma, can directly affect centrosomal structure and spindle integrity and potentially modulate apoptotic and cell cycle progression pathways.     

TPX2 knockdown suppressed hepatocellular carcinoma cell invasion via inactivating AKT signaling and inhibiting MMP2 and MMP9 expression

Objective： Targeting protein for Xenopus kinesin-like protein 2 （TPX2） is a nuclear proliferation-related protein that plays a critical role in the formation of mitotic spindle. High expression of TPX2 has been observed in several types of tumors. However, the role of TPX2 in hepatocellular carcinoma （HCC） remains unclear. Our study aimed to investigate the effect of TPX2 on HCC cell invasion. Methods： The immortalized normal human liver cell line L02 and six HCC cell lines including SMMC- 7721, BEL-7402, Huh-7, HepG2, Hep3B and SKHepl were subjected to qRT-PCR and western blot for TPX2 mRNA and protein, respectively. Furthermore, TPX2 small interfering RNA （siRNA） was used to knock down TPX2 expression in SMMC-7721 and HepG2 cells. Cell proliferation and invasion were determined by MTT and transwell assays. Otherwise, expression of p-AKT, MMP2 and MMP9 were evaluated by western blot in SMMC-7721 cells. Results： The expression of TPX2 in HCC cell lines was markedly higher than that in normal human liver cell line. TPX2 knockdown using a specific TPX2-siP, NA reduced the number of invaded cells and inhibited cell proliferation in SMMC-7721 and HepG2 cells. Furthermore, TPX2 knockdown resulted in inactivation of AKT signaling and down-regulation of MMP2 and MMP9 expression in SMMC-7721 cells. Conclusions： Our study identified that TPX2 might contribute to tumor cell invasion through activating AKT signaling and subsequently increasing MMP2 and MMP9 in HCC.     

慢病毒介导的TPX2基因沉默对人宫颈癌HeLa细胞系增殖、迁移和细胞周期的影响及机制

目的：研究慢病毒介导的TPX2基因沉默对人宫颈癌HeLa细胞系增殖、迁移和细胞周期的影响及其机制。方法：构建4种靶向TPX2基因的慢病毒表达载体（LV-TPX2-shRNA-1/2/3/4）,同时构建阴性对照质粒,将5种质粒分别转染到293T细胞中制备慢病毒。慢病毒感染宫颈癌HeLa细胞后,实时荧光定量PCR和Western blot分别检测TPX2 mRNA和蛋白的沉默效果。选择沉默效果最佳的重组慢病毒进行后续功能实验。分别采用CCK-8法、Transwell迁移实验及流式细胞术检测各组细胞的增殖、迁移及细胞周期分布情况。Western blot检测TPX2-shRNA转染前后Ki-67、cyclin B2、Aurora-A及P53的表达。结果：与对照组比较,构建的靶向TPX2基因的RNA干扰慢病毒载体均可持续稳定的抑制HeLa细胞TPX2基因的表达,尤其以LV-TPX2-shRNA-1最为明显（P < 0.01）,故选择LV-TPX2-shRNA-1进行后续实验。与对照组比较,沉默TPX2基因的表达能降低HeLa细胞增殖及迁移能力（P < 0.05）,使G2及S期细胞比例明显增加（P < 0.05）,且上调P53蛋白的表达水平,下调Ki-67、cyclin B2及Aurora-A蛋白的表达水平（P均 < 0.05）。结论：沉默TPX2基因蛋白表达能抑制宫颈癌细胞的增殖及迁移能力,可能与其改变细胞周期分布及下调Ki-67、cyclin B2、Aurora-A蛋白表达水平,上调P53蛋白表达水平有关。