同源异型盒基因HOXA5对胃癌细胞BGC823侵袭能力的影响

目的 检测人胃癌组织中HOXA5基因的表达情况,观察干扰HOXA5基因的表达之后,胃癌细胞BGC823的迁移和侵袭能力的变化,为阐明HOXA5在胃癌向远处转移的分子机制中提供新的思路.方法 通过实时定量PCR(qRT-PCR)、Western blot以及免疫组织化学等技术,检测胃癌组织中HOXA5基因的表达.应用细胞转染的方法 将HOXA5基因的过表达质粒转入胃癌细胞BGC823中,经qRT-PCR和Western blot验证HOXA5表达上调之后,利用Transwell侵袭实验检测BGC823细胞侵袭能力的变化.结果 在收集的69例新鲜的胃癌组织以及相对应的癌旁组织中,免疫组织化学显示相比于癌旁组织的表达情况,胃癌组织中只有26例HOXA5的表达高于癌旁组织,比例为37.7％;qRT-PCR、Western blot检测发现相对于癌旁组织,胃癌组织中HOXA5的表达水平显著下调(mRNA水平下调3倍,蛋白水平下调2倍,差异具有统计学意义,P<0.05).BGC823细胞转染了过表达质粒之后,HOXA5的表达能够被上调.过表达了HOXA5基因的BGC823细胞的侵袭能力下降.结论 在胃癌组织中HOXA5的表达下调,而在胃癌细胞BGC823中增强HOXA5的表达能够降低细胞的侵袭能力,说明HOXA5在胃癌的发生发展和侵袭、转移中发挥着抑癌基因的作用,可作为一个新的治疗靶点.     

A new recurrent inversion, inv(7)(p15q34), leads to transcriptional activation of HOXA10 and HOXA11 in a subset of T-cell acute lymphoblastic leukemias.

Chromosomal translocations with breakpoints in T-cell receptor (TCR) genes are recurrent in T-cell malignancies. These translocations involve the TCRalphadelta gene (14q11), the TCRbeta gene (7q34) and to a lesser extent the TCRgamma gene at chromosomal band 7p14 and juxtapose T-cell oncogenes next to TCR regulatory sequences leading to deregulated expression of those oncogenes. Here, we describe a new recurrent chromosomal inversion of chromosome 7, inv(7)(p15q34), in a subset of patients with T-cell acute lymphoblastic leukemia characterized by CD2 negative and CD4 positive, CD8 negative blasts. This rearrangement juxtaposes the distal part of the HOXA gene cluster on 7p15 to the TCRbeta locus on 7q34. Real time quantitative PCR analysis for all HOXA genes revealed high levels of HOXA10 and HOXA11 expression in all inv(7) positive cases. This is the first report of a recurrent chromosome rearrangement targeting the HOXA gene cluster in T-cell malignancies resulting in deregulated HOXA gene expression (particularly HOXA10 and HOXA11) and is in keeping with a previous report suggesting HOXA deregulation in MLL-rearranged T- and B cell lymphoblastic leukemia as the key factor in leukaemic transformation. Finally, our observation also supports the previous suggested role of HOXA10 and HOXA11 in normal thymocyte development.     

Predominant Rab-GTPase amplicons contributing to oral squamous cell carcinoma progression to metastasis

Metastatic oral squamous cell carcinoma (OSCC) is frequently associated with recurrent gene abnormalities at specific chromosomal loci. Here, we utilized array comparative genomic hybridization and genome-wide screening of metastatic and non-metastatic tongue tumors to investigate genes potentially contributing to OSCC progression to metastasis. We identified predominant amplifications of chromosomal regions that encompass the RAB5, RAB7 and RAB11 genes (3p24-p22, 3q21.3 and 8p11–12, respectively) in metastatic OSCC. The expression of these Rab GTPases was confirmed by immunohistochemistry in OSCC tissues from a cohort of patients with a follow-up of 10 years. A significant overexpression of Rab5, Rab7 and Rab11 was observed in advanced OSCC cases and co-overexpression of these Rabs was predictive of poor survival (log-rank test, P = 0.006). We generated a Rab interaction network and identified central Rab interactions of relevance to metastasis signaling, including focal adhesion proteins. In preclinical models, mRNA and protein expression levels of these Rab members were elevated in a panel of invasive OSCC cell lines, and their down-regulation prevented cell invasion at least in part via inhibition of focal adhesion disassembly. In summary, our results provide insights into the cooperative role of Rab gene amplifications in OSCC progression and support their potential utility as prognostic markers and therapeutic approach for advanced OSCC.     

基于乳腺原发癌基因表达谱的预后分子分型研究

目的比较乳腺原发癌随访3年有远处转移病例与无远处转移病例的原发癌基因表达谱差异,筛选转移预后预测标志基因群,并探讨其用于乳腺癌患者预后分子分型的临床意义。方法以随访3年5例有远处转移和5例无远处转移的乳腺癌患者作为分析病例,采用含21329个人功能基因的Oligo芯片筛选预后不同的乳腺原发癌组织差异表达基因;基于差异表达基因分别对10例分析病例和另外20例验证病例进行k-mean聚类分析;利用逐一剔除法去除对聚类没有贡献的基因,得到可以预测乳腺癌患者转移和预后的标志基因群,用于乳腺癌患者预后的分子分型。结果利用10例不同预后的分析病例筛选得到的差异表达基因可以将30例乳腺癌患者分为预后良好和预后不良两个组;随访3年有远处转移的病例全部聚类在预后不良组,占该组的70．0％（7／10）,而预后良好组中不含有远处转移的病例（0／20）,两组3年内发生远处转移率的差异有统计学意义（P=0．03）。由104个基因组成的预后预测基因群中,5个与细胞黏附运动相关的基因在预后不良组较预后良好组均表达上调,2个与机体免疫应答相关的基因均下调,11个与细胞生长代谢相关的基因上调,14个与细胞生长代谢相关的基因下调,15个与细胞信号传导相关的基因表达量显著改变。结论通过乳腺癌有远处转移病例与无远处转移病例原发癌基因表达谱的比较研究,筛选得到可以预测乳腺癌患者转移和预后的基因群,且可反映高转移能力的乳腺原发癌中肿瘤细胞运动能力增强、机体免疫识别缺陷、细胞增殖活跃和信号传导异常的生物学特征,并能对乳腺癌患者进行预后分子分型,有望用于临床指导个体化治疗方案的选择。     

Positive correlations of Oct-4 and Nanog in oral cancer stem-like cells and high-grade oral squamous cell carcinoma.

PURPOSE: Oral squamous cell carcinoma (OSCC), like many solid tumors, contains a heterogeneous population of cancer cells. Recent data suggest that a rare subpopulation of cancer cells, termed cancer stem cells (CSC), is capable of initiating, maintaining, and expanding the growth of tumor. Identification and characterization of CSC from OSCC facilitates the monitoring, therapy, or prevention of OSCC. EXPERIMENTAL DESIGN: We enriched oral cancer stem-like cells (OC-SLC) through sphere formation by cultivating OSCC cells from established OSCC cell lines or primary cultures of OSCC patients within defined serum-free medium. Differential expression profile of stemness genes between enriched OC-SLC and parental OSCC was elucidated. Furthermore, immunohistochemical staining of stemness markers on OSCC patient tissues was examined to evaluate the association between stemness genes and prognosis of OSCC. RESULTS: Enriched OC-SLC highly expressed the stem/progenitor cell markers and ABC transporter gene (Oct-4, Nanog, CD117, Nestin, CD133, and ABCG2) and also displayed induced differentiation abilities and enhanced migration/invasion/malignancy capabilities in vitro and in vivo. Elevated expression of CD133 was shown in the enriched OC-SLC from OSCC patients' tumors. Positive correlations of Oct-4, Nanog, or CD133 expression on tumor stage were shown on 52 OSCC patient tissues. Kaplan-Meier analyses exhibited that Nanog/Oct-4/CD133 triple-positive patients predicted the worst survival prognosis of OSCC patients. CONCLUSION: We enriched a subpopulation of cancer stem-like cell from OSCC by sphere formation. The enriched OC-SLC possesses the characteristics of both stem cells and malignant tumors. Additionally, expression of stemness markers (Nanog/Oct-4/CD133) contradicts the survival prognosis of OSCC patients.     

Integrated Bioinformatics Analysis of mRNAs and miRNAs Identified Potential Biomarkers of Oral Squamous Cell Carcinoma

Background: Oral cancer is a frequently encountered neoplasm of the head and neck region, being the eighth most common type of human malignancy worldwide. Despite improvement in its control, morbidity and mortality, rates have improved little in the past decades. The present investigations about gene interaction and pathways still could not clear the appearance and development of oral squamous cell carcinoma (OSCC), completely. The aim of this study is to investigate the key genes and microRNAs interaction in OSCC. Materials and Methods: The microarray datasets GSE13601 and GSE98463, including mRNA and miRNA profiles, were extracted from the GEO database and were analyzed using GEO2R. Functional and pathway enrichment analyses were performed by using the DAVID database. The protein–protein interaction (PPI) network was constructed and analyzed using STRING database and Cytoscape software, respectively. Finally, miRDB was applied to predict the targets of the differentially expressed miRNAs (DEMs). Results: Totally, 97 differentially expressed genes (DEGs) were found in OSCC, including 66 up-regulated and 31 down-regulated genes. The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses showed that up-regulated genes were significantly enriched in movement of cell or subcellular component, cell adhesion, biological adhesion, cellular localization, apoptotic signaling pathway, while the down-regulated genes were enriched in muscle system process and oxidation-reduction process. From the PPI network, the top 10 nodes with the highest degree were detected as hub genes. In addition, 18 DEMs were screened, which included 7 up-regulated and 11 down-regulated miRNAs. STAT1 was potentially targeted by three miRNAs, including has-miR-6825-5P, has-miR-4495, and has-miR-5580-3P. Conclusion: The roles of DEMs such as hsa-mir-5580-3p in OSCC through interactions with DEGs CD44, ACLY, ACTR3, STAT1, LAMC2 and YWHAZ may offer a suitable candidate biomarker pattern for diagnosis, prognosis and treatment processes in OSCC.     

HOXA5基因与肿瘤

HOXA5作为同源盒(HOX)基因家族的一员,在多器官中都有表达,具有调节基因表达、细胞分化和机体形态发生的功能,其结构和(或)功能异常与白血病、乳腺癌、脑血管瘤、肝细胞癌等肿瘤的发生密切相关.研究HOXA5与肿瘤的关系有助于肿瘤的诊断、治疗及预防.     

HOXA10 基因在子宫内膜癌组织中的表达及其生物学作用

目的:探讨同源框A10（HOXA10）基因在子宫内膜癌组织中的表达及其对子宫内膜癌Ishikawa 细胞株凋亡、迁移及侵袭的影响。方法：收集2012 年至2013 年在鼓楼医院妇产科子宫内膜癌组织标本21 例、正常增殖期子宫内膜组织标本25 例,采用实时荧光定量PCR(qRT-PCR)及Western blotting 方法检测HOXA10 在子宫内膜癌组织及正常子宫内膜组织中的表达。将感染复数为5、10、20 MOI的ad-flag-HOXA10 腺病毒质粒及20 MOI的ad-flag-lacz 腺病毒质粒(对照组)感染人子宫内膜癌Ishikawa 细胞株,流式细胞术检测各组细胞凋亡情况。将50 nmol/L的si-HOXA10 及si-NC质粒转染Ishikawa 细胞株,分别为下调组及下调对照组;将20 MOI的ad-flag-HOXA10 及20 MOI的ad-flag-lacz 腺病毒质粒感染Ishikawa 细胞株,分别为上调组及上调对照组;Transwell 小室检测各组细胞迁移及侵袭能力。结果：在子宫内膜癌组织中HOXA10 mRNA 表达量比正常子宫内膜组织中显著降低[ （0.56±0.14）vs (1.36±0.33),P<0.01],其蛋白表达量同样显著降低[ （1.01±0.25）vs (2.10±0.71),P<0.01]。上调HOXA10后5、10、20 MOI 组细胞的凋亡率明显升高,且大多数处于早期凋亡[ （50.92±8.79）%、(55.17±4.07)%、(76.10±3.65)% vs (7.74±0.15)%,均P<0.01]。下调HOXA10 表达后迁移细胞数显著增加[ （248±25）vs (135±15)个,P<0.01],上调HOXA10 表达后迁移细胞数显著减少[（50±6）vs (100±13) 个,P<0.01];下调HOXA10 表达后侵袭细胞数显著增多[ （131±18）vs (66±9)个,P<0.01],上调HOXA10 表达后侵袭细胞数显著减少[ （34±8）vs（60±4）个,P<0.01]。结论：HOXA10 基因在子宫内膜癌内膜中表达低于正常内膜,子宫内膜癌Ishikawa细胞株中上调HOXA10 基因表达能促进细胞凋亡并抑制其迁移和侵袭能力。     

MEIS1 and HOXA7 genes in human acute myeloid leukemia

Co-activation of Meisl with Hoxa7 or Hoxa9 homeobox genes by retroviral gene insertion has recently been reported to be leukemogenic in murine myeloid leukemia. In this study we determined their expression in human leukemia. Most human myeloid leukemia cell lines co-expressed MEIS1 with HOXA7 and HOXA9. Among patients with acute leukemia, 50% of AML patients expressed MEIS1, while the majority of ALL patients were negative. A total of 89.5% of patients expressing MEIS1 co-expressed HOXA7. In unadjusted models, poorer response to chemotherapy was associated with expression of HOXA7 regardless of MEIS1 status and older patients were more likely to express either gene.