cDNA芯片在卵巢癌早诊及临床病理分期中的初步应用探讨

目的:探讨PTEN,nm23H₁,VEGF₁₆₅,Tiam1,MMP-2,Timp2,HE4和S100A4基因在卵巢癌发生和侵袭中的作用。方法:采用cDNA芯片技术分别检测20例卵巢癌和5例正常卵巢组织中上述基因cDNA表达谱差异。结果:PTEN,Timp2和nm23H₁ cDNA在卵巢癌组织中表达下调(CY-3/CY-5<0.5);Tiam1,VEGF₁₆₅,MMP-2,HE4和S100A4cDNA在卵巢癌组织中表达上调(CY-3/CY-5>2.0);且HE4基因表达上调的最为明显(CY-3/CY-5>5.0)。表达下调和表达上调的上述基因与卵巢癌临床病理分期、组织分化程度关系密切(P<0.01)。结论:上述基因与卵巢癌发生及侵袭关系密切。cDNA基因芯片技术对于探讨卵巢癌分子机制,寻找卵巢癌分子标志物具有重要意义。     

乳腺癌的早期预警

绝经期前后是女性乳腺癌的高发年龄阶段.CA-153是临床上常用的监测乳腺癌的肿瘤标记物,但在早期诊断中却无明显意义.而基因表达谱技术成为目前研究热点,并研究发现了乳腺癌相关基因.而临床用于诊断乳腺癌的方法也存在一定弊端,因此对于提前预测或早期发现乳腺癌尚无确切定论.     

肺癌的早期诊断

肺癌已成为人类癌症死亡的主要原因之一,5年生存率仅10％。既往应用胸部X线和痰细胞学进行筛查和早期诊断的研究没有达到诊断筛查的目的－－肺癌死亡率的下降。随着新技术的发展,人们对肺癌筛查重新产生了兴趣。本文回顾了肺癌早期诊断最新进展,有低剂量CT、液基细胞学技术、荧光内镜和分子病理学等方面。     

使用cDNA微阵列和组织微阵列对三种上皮性卵巢肿瘤基因表达的分析

背景与目的：卵巢癌是在妇科恶性肿瘤中死亡率最高的肿瘤,主要原因是由于缺乏有效的早期诊断方法。为了发现新的、特异性癌基因和进一步探索上皮性卵巢癌的临床意义,本文探索出一种新方法,通过结合cDNA微阵列和RNA原位杂交冰冻组织微阵列的方法寻找特异性卵巢癌基因。方法：利用cDNA微阵列筛选在3种不同卵巢癌中(卵巢浆液性交界性肿瘤、卵巢浆液性腺癌和卵巢子宫内膜样腺癌)显示有意义表达的基因,并由RNA原位杂交冰冻组织微阵列证实其结果。结果：40个基因和ESTs显示出在卵巢浆液性交界性肿瘤、浆液性和子宫内膜样卵巢癌3种类型之间基因表达有明显的差异;EPHB6、PrrPRF、GFER、ERG25、PLRP1,FLJ22060和WISP2被进一步用于RNA原位杂交冰冻组织微阵列研究,其结果与cDNA微阵列研究结果相符合。结论：cDNA微阵列和RNA原位杂交冰冻组织微阵列相结合是一种理想的寻找癌相关基因的方法,EPHB6,PrPRF,GFER,ERG25,PLRP1,FL122060和WISP2有可能成为新的上皮性卵巢癌候选基因。     

A distinct molecular profile associated with mucinous epithelial ovarian cancer

Mucinous epithelial ovarian cancers (MOC) are clinically and morphologically distinct from the other histological subtypes of ovarian cancer. To determine the genetic basis of MOC and to identify potential tumour markers, gene expression profiling of 49 primary ovarian cancers of different histological subtypes was performed using a customised oligonucleotide microarray containing >59 000 probesets. The results show that MOC express a genetic profile that both differs and overlaps with other subtypes of epithelial ovarian cancer. Concordant with its histological phenotype, MOC express genes characteristic of mucinous carcinomas of varying epithelial origin, including intestinal carcinomas. Differences in gene expression between MOC and other histological subtypes of ovarian cancer were confirmed by RT-PCR and/or immunohistochemistry. In particular, galectin 4 (LGALS4) was highly and specifically expressed in MOC, but expressed at lower levels in benign mucinous cysts and borderline (atypical proliferative) tumours, supporting a malignant progression model of MOC. Hence LGALS4 may have application as an early and differential diagnostic marker of MOC.     

胰腺癌的早期诊断

胰腺癌的早期症状和体征多无特异性,现有的辅助检查及肿瘤标志物并未能使胰腺癌的早期诊断率大幅度提高。复习相关资料,就流行病学,影像学和肿瘤标志物三方面来评价目前胰腺癌早期诊断的现状及进展作一综述。     

cDNA微阵列对22例弥漫型胃癌基因的检测

目的从转录组水平识别弥漫型胃癌与正常胃黏膜间的基因表达差异,探讨胃癌分子发生发展机制。方法收集22例弥漫型胃癌患者的新鲜冻存胃癌组织及同例相应正常胃黏膜。杂交芯片采用含14592个点的cDNA表达谱芯片。差异表达基因的筛选标准为该基因在50％以上样本中的肿瘤与正常组织荧光强度比（ratio比值）〉2或〈0．5。采用系统聚类法进行基因表达的相似性分析,标本组间比较采用方差分析。应用实时定量RT—PCR方法对芯片结果进行验证。结果胃癌组织与正常胃黏膜间的差异表达基因共357个,其中表达上调者153个,下调者204个。上调基因功能主要与细胞骨架运动、基质重建、细胞增殖及信号传导等相关;下调基因功能则主要与细胞免疫防御、毒理代谢、功能分化、核一浆转运及凋亡抑制等相关。TNM分期的Ⅰ＋Ⅱ期组与Ⅲ＋Ⅳ期组间,有7个基因的表达差异有统计学意义（P〈0．05）。RT—PCR验证结果与芯片表达结果一致。结论运用cDNA芯片进行弥漫型胃癌基因表达谱分析,有助于从分子水平全方位阐明弥漫型胃癌的发病机制及生物学特性,也有助于进一步发现新的分子诊断指标和基因治疗靶标。     

A CLINICAL STUDY ON PROSTATE CANCER DIAGNOSIS WITH cDNA MACROARRAY

Objective: To diagnose prostatic cancer (CAP) with cDNA macroarray. Methods: Total RNA was isolated from patients with prostate cancer and from normal people, and poly(A) RNA was further purified. Then differentially expressed genes were analysed in CaP and normal prostate by cDNA macroarray system. Results: There were differential expressions of nine prostate-associated specific genes in CaP as compared with normal prostate, among which, 7 were significantly up-regulated and 2 were down-regulated. Conclusion: As a diagnostic approach at molecular level, the cDNA macroarray is supposed to elevate the detection rate of CaP.     

利用基因芯片技术研究薯蓣皂苷抗乳腺癌的作用机制

目的:通过基因芯片技术研究薯蓣皂苷对乳腺癌细胞基因表达的影响,初步探索药物作用的靶基因。方法:提取总RNA,通过反转录方法制备cDNA荧光探针,进行芯片杂交,然后对芯片进行图像扫描,最后用ImaGene软件包对荧光信号进行分析。结果:57个有信号的基因克隆中,用药前28个高表达,用药后仅3个基因克隆高表达,28个表达无明显差别,低表达基因克隆26个。结论:薯蓣皂苷干预后,引起细胞-met、HGF/SF、myb和CDK4基因表达丰度下降,提示薯蓣皂苷抗肿瘤细胞增殖可能与抑制细胞-met、HGF/SF、myb基因和CDK4基因表达相关。     

正常卵巢上皮细胞和卵巢上皮癌细胞基因表达差异的研究

目的：探讨卵巢上皮癌细胞相关基因的差异表达。方法：采用舍384条肿瘤相关基因的cDNA阵谱检测了卵巢上皮癌细胞株SKOV-3及正常卵巢上皮细胞的基因谱，分析卵巢上皮癌细胞相关基因的差异表达。结果：在384条候选基因中，与卵巢癌相关的差异表达基因33条，其中22条表达上调，11条表达下调。结论：cDNA阵谱技术是筛查卵巢癌相关基因的有效方法。     

Analysis of Metastatic-Related Gene Expression in Gastric Cancer by Low-Density cDNA Microarrays

OBJECTIVE To screen metastatic-related genes in human gastric cancer by a low-density cDNA microarray technique. METHODS A total of 18 paired gastric cancer and adjacent normal mu-cosa were examined by a low-density cDNA microarray containing 23 genes. RT-PCR was used for further verification. RESULTS The mRNA expression of MMP -7, heparanase, S100A4, hTERT, hRad17 in gastric cancers was higher than that in coupled normal mucosa (P=0.002, 0.00011, 0.000072, 0.002, 0.00016 respectively), whereas nm23H1, and CDH1 were lower (P=0.003, 0.012 respectively). The concordance was verified further by RT-PCR with a correlation coefficient of 0.774. In gastric primary lesions the mRNA expression of MMP-7, heparanase and S100A4 was higher in the serosa involved compared to non-involved (P=0.003, 0.009, 0.012 respectively), whereas nm23H1, CDH1, KAI1 were lower (P=0.001, 0.001, 0.006 respectively). With respect to the area of serosa involvement, MMP-7 and heparanase expressions were higher in an area of more than 20 cm2 compared to an area of less than 20 cm2 (P=0.001, 0.02 respectively), whereas nm23H1, CDH1 and KAI1 were lower (P=0.030, 0.041, 0.031 respectively). MMP-7 and hTERT expressions were higher in the heavier lymph node metastatic cases (no less than 7) than in the lighter lymph node metastatic cases (no more than 6, P=0.001, 0.005 respectively). CONCLUSION Expression of MMP -7, S100A4, heparanase, hTERT, KAI1, CDH1 and nm23H1 correlated closely with invasion and metastasis in gastric carcinomas. The low-density cDNA microarrays can be used to examine the expression of many genes simultaneously, parallely and quickly.     

Molecular profiling of platinum resistant ovarian cancer

The aim of this study is to discover a gene set that can predict resistance to platinum-based chemotherapy in ovarian cancer. The study was performed on 96 primary ovarian adenocarcinoma specimens from 2 hospitals all treated with platinum-based chemotherapy. In our search for genes, 24 specimens of the discovery set (5 nonresponders and 19 responders) were profiled in duplicate with 18K cDNA microarrays. Confirmation was done using quantitative RT-PCR on 72 independent specimens (9 nonresponders and 63 responders). Sixty-nine genes were differentially expressed between the nonresponders (n=5) and the responders (n=19) in the discovery phase. An algorithm was constructed to identify predictive genes in this discovery set. This resulted in 9 genes (FN1, TOP2A, LBR, ASS, COL3A1, STK6, SGPP1, ITGAE, PCNA), which were confirmed with qRT-PCR. This gene set predicted platinum resistance in an independent validation set of 72 tumours with a sensitivity of 89% (95% CI: 0.68-1.09) and a specificity of 59% (95% CI: 0.47-0.71)(OR=0.09, p=0.026). Multivariable analysis including patient and tumour characteristics demonstrated that this set of 9 genes is independent for the prediction of resistance (p<0.01). The findings of this study are the discovery of a gene signature that classifies the tumours, according to their response, and a 9-gene set that determines resistance in an independent validation set that outperforms patient and tumour characteristics. A larger independent multicentre study should further confirm whether this 9-gene set can identify the patients who will not respond to platinum-based chemotherapy and could benefit from other therapies.     

乳腺癌早期诊断技术

乳腺癌的早期诊断对于改善患者预后至关重要.随着新型诊断技术的出现,乳腺癌早期诊断率不断提高.体液中的微小RNA (miRNA)、唾液蛋白、尿液多胺作为新型肿瘤标志物其敏感性和特异性均较高;传统的影像学诊断技术在光学技术和计算机的辅助下也有较大突破;影像引导微创活检技术如空芯针活检和真空辅助活检,具有准确、经济、创伤小等优点,并能提供更多的标本,已广泛用于临床.     

Hypodontia phenotype in patients with epithelial ovarian cancer

Background

Ovarian cancer is usually diagnosed in an advanced stage and the present clinical and diagnostic molecular markers for early OC screening are insufficient. The aim of this study was to identify potential relationship between the hypodontia and epithelial ovarian cancer (EOC).

Patients and methods

A retrospective study was conducted on 120 patients with EOC treated at the Department of Gynaecologic and Breast Oncology at the University Clinical Centre and 120 gynaecological healthy women (control group) of the same mean age. Women in both groups were reviewed for the presence of hypodontia and the patients with EOC also for clinicopathological characteristics of EOC according to hypodontia phenotype.

Results

Hypodontia was diagnosed in 23 (19.2%) of patients with EOC and 8 (6.7%) controls (p = 0.004; odds ratio [OR] = 3.32; confidence interval [CI], 1.42–7.76). There was no statistically significant difference in patients with EOC with or without hypodontia regarding histological subtype (p = 0.220); they differed in regard to FIGO stage (p = 0.014; OR =3.26; CI, 1.23–8.64) and tumour differentiation grade (p = 0.042; OR = 3.1; CI, 1.01–9.53). Also, bilateral occurrence of EOC was more common than unilateral occurrence in women with hypodontia (p = 0.021; OR = 2.9; CI, 1.15–7.36). We also found statistically significant difference between the ovarian cancer group and control group in presence of other malignant tumours in subjects (p < 0.001).

Conclusions

The results of the study suggest a statistical association between EOC and hypodontia phenotype. Hypodontia might serve as a risk factor for EOC detection.     

基因表达谱芯片筛选甲状腺癌失分化相关基因的初步研究

目的应用基因芯片技术筛查甲状腺癌失分化相关基因,为进一步研究其发病机制提供新的思路和线索。方法分别用Cy5和Cy3两种不同的荧光染料通过逆转录反应将失分化甲状腺癌组和对照组分化型甲状腺癌组织的mRNA分别标记成探针,并与载有一组靶基因的基因表达谱芯片进行杂交。通过扫描荧光强度,计算机软件分析,寻找两组差异表达基因。结果分化型甲状腺癌与失分化甲状腺癌组织之间存在一系列差异表达基因,共有373条差异表达基因,其中表达增加的有209条(2.0倍以上),表达降低的有164条(0.5倍以下)。结论基因芯片为筛选甲状腺癌失分化相关基因提供了有效的方法。     

胰腺癌的早期诊断

胰腺癌是一种恶性程度高、预后较差的消化系统肿瘤。如果胰腺癌患者早期能够得到外科治疗,其生存状况将会明显改善。早期发现、早期诊断和早期治疗是提高存活率的关键。人们一直在努力寻找早期诊断胰腺癌的标记物如肿瘤血清学标记物、基因标记物,其中血清学标记物中有MUC基因、CA199、CA242和PSGF、CAM17．1;基因诊断标记物有癌基因、抑癌基因和端粒酶。同时影像学的发展也在早期诊断胰腺癌方面发挥了一定作用。     

胰腺癌的早期诊断

胰腙癌是一种恶性程度高、预后较差的消化系统肿瘤。如果胰腙癌患者早期能够得到外科治疗，其生存状况将会明显改善。早期发现、早期诊断和早期治疗是提高存活率的关键。人们一直在努力寻找早期诊断胰腺癌的标记物如肿瘤血清学标记物、基因标记物，其中血清学标记物中有MUC基因、CA199、CA242和TSGF、CAM17．1；基因诊断标记物有癌基因、抑癌基因和端粒酶。同时影像学的发展也在早期诊断胰腺癌方面发挥了一定作用。     

Identification of differentially expressed genes in gastric cancer by high density cDNA microarray

The identification of molecular markers for diagnosis, treatment, and prognosis is a significant issue in the management of patients with gastric cancer. We compared the expression profiles of 23 gastric cancers and 22 normal gastric tissues using cDNA microarrays. We divided the samples into two sets, 11 pairs as a training set and 12 unpaired gastric cancer and 11 unpaired normal gastric tissues as a test set. We selected significant genes in the training set and validated the significance of the genes in the test set. We obtained 238 classifier genes that showed a maximum cross-validation probability and clear hierarchical clustering pattern in the training set, and showed excellent class prediction probability in the independent test set. The classifier genes consisted of known genes related to the biological features of cancer and 28% unknown genes. We obtained genome-wide molecular signatures of gastric cancer, which provides preliminary exploration data for the pathophysiology of gastric cancer.     

应用微矩阵表达谱基因芯片筛选食管癌新的相关基因

[目的]应用微矩阵表达谱基因芯片筛选食管癌新的相关基因.[方法]微矩阵表达谱基因芯片(14114种基因)筛选3例食管癌及其对照癌旁组织的差异表达基因,用Northern印迹证实,用末端终止法测序,将测序结果在GenBank数据库进行同源性检索.[结果]检测的3例临床标本中,共有的差异表达基因31条,上调基因10条,下调基因21条,其序列与Genbank数据库比较,从中筛选出2条无明显同源的人类已知基因或片段,即新基因,并列出其序列.[结论]微矩阵表达谱基因芯片可应用于筛选食管癌新的相关基因,其具备高通量、特异性好、快速的特点;食管癌和对照癌旁组织间存在2条新的差异表达基因.