骨肉瘤组织MGMT基因甲基化状态的检测及其临床意义分析

目的探讨骨肉瘤组织O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)基因甲基化状态与MGMT蛋白表达及骨肉瘤化疗后肿瘤坏死率的关系。方法收集51例骨肉瘤组织样本,用甲基化特异性PCR(MSP)方法检测MGMT基因启动子区的甲基化状态,免疫组化方法检测MGMT蛋白的表达,并分析MGMT基因甲基化状态与肿瘤坏死率及临床病理资料的关系。结果 51例骨肉瘤组织MGMT基因启动子区甲基化发生率为23.5%(12/51),MGMT蛋白表达阴性率为27.5%(14/51);MGMT基因甲基化状态与骨肉瘤患者的年龄、性别、肿瘤大小、肿瘤类型等无明显关系(P0.05)。MGMT基因甲基化的患者(共12例)化疗后肿瘤坏死率为Ⅰ级者0例,Ⅱ级者2例(16.7%),Ⅲ级者3例25.0%(25.0%),Ⅳ级者7例(58.3%),MGMT基因未甲基化的患者(共39例)化疗后肿瘤坏死率为Ⅰ级者23例(59.0%),Ⅱ级者13例(33.3%),Ⅲ级者2例(5.1%),Ⅳ级者1例(2.6%)。Wilcoxon秩和检验显示,骨肉瘤组织MGMT基因未甲基化患者的MGMT蛋白表达明显高于甲基化的患者(u=-4.92,P0.001),而化疗后肿瘤坏死率明显低于甲基化的患者(P0.05)。结论骨肉瘤组织MGMT基因甲基化状态对判断骨肉瘤患者化疗后的肿瘤坏死率具有重要意义。     

肺癌患者CDH13基因启动子甲基化状态研究

目的应用甲基化特异性PCR（MSP）方法检测非小细胞肺癌（NSCLC）CDH13基因启动子甲基化情况。方法留取44例非小细胞肺癌患者手术标本及12例非肺癌患者正常肺组织,MSP分析CDH13基因启动子甲基化情况。结果 44例非小细胞肺癌中CDH13基因启动子甲基化阳性率为54.5%（24/44）,12例正常肺组织中未检测到此基因的甲基化（0/12）。CDH13基因甲基化与患者性别及吸烟与否无关;肿瘤分期越晚甲基化的发生率越高。结论原发性非小细胞肺癌中存在着较高比例的CDH13启动子甲基化,提示CDH13在非小细胞肺癌的发生中起着重要作用。     

乳腺癌发生、发展过程中相关基因甲基化的研究现状

DNA甲基化是指生物体在DNA甲基转移酶的催化下,以S-腺苷甲硫氨酸为甲基供体,将甲基转移到特定的碱基上的过程.在哺乳动物,DNA甲基化主要发生在5'-CpG-3'的C上,生成5-甲基胞嘧啶.甲基化状态的改变是导致肿瘤发生的一个重要因素.启动子区低甲基化导致原癌基因及其他肿瘤相关基因的转录激活,从而促进肿瘤的发生发展[1].     

细胞周期蛋白依赖性激酶抑制因子3在宫颈癌中表达及临床意义

目的 探讨细胞周期蛋白依赖性激酶抑制因子3(CDKN3)在宫颈癌中的表达及临床意义.方法 收集自2015年1月至2016年1月于中国医科大学肿瘤医院妇科行手术治疗且经病理检查确诊患者的60例宫颈癌组织、30例宫颈原位癌组织、30例正常宫颈组织.免疫组织化学法检测宫颈癌组织、宫颈原位癌组织、正常宫颈组织CDKN3的表达水...     

HOXA10基因启动子区5'CpG岛甲基化在子宫内膜异位症中的作用

目的 检测HOXA10基因启动子区5'CpG岛甲基化状况,探讨其在子宫内膜异位症(EMS)发生中的作用.方法 收集2007年9月-2008年5月南方医科大学珠江医院妇产科30例EMS患者作为EMS组,同期25例正常妇女作为对照组,采用甲基化特异性聚合酶链反应(MSP)检测EMS组异位内膜组织(其中Ⅰ-Ⅱ期10例,Ⅲ-Ⅳ期20例)和对照组子宫内膜组织中HOXA10基因启动子区5'CpG岛甲基化情况,并比较其在不同EMS临床分型间的差异.结果 30例EMS患者异位内膜组织中检测到程度不等的甲基化,其中20例Ⅲ-Ⅳ期EMS患者中有11例发生甲基化,甲基化发生率为55%,10例Ⅰ-Ⅱ期EMS患者中有1例发生甲基化,甲基化发生率为10%,Ⅲ-Ⅳ期EMS患者异位内膜组织HOXA10基因启动子区5'CpG岛的甲基化发生率高于Ⅰ-Ⅱ期(P<0.05).25例正常子宫内膜组织均未检测到甲基化.结论 HOXA10基因启动子区5'CpG岛甲基化在EMS的发生、发展中可能起着重要作用.     

肺癌组织中p16基因的异常甲基化分析

DNA甲基化具有多方面的生物学意义,它是由甲基转移酶介导,以S-腺苷-L-甲硫氨酸为甲基供体,在胞嘧啶5位碳原子上加入一甲基基团,DNA甲基化是最常见的复制后调节方式之一,在基因表达、胚胎发育、基因组印迹等方面发挥重要作用^[1]。     

鼻咽癌中hMLH1基因启动子甲基化状态及转录表达的研究

目的 探讨鼻咽癌组织中hMLH1基因启动子区甲基化状态及其对转录表达的影响.方法 运用甲基化特异性FCR和半定量反转录PCR法检测54例鼻咽癌组织中hMLH1基因启动子甲基化状态及其转录表达水平,分析鼻咽癌组织中hMLH1甲基化与临床资料的关系,以及hMLH1甲基化对转录表达的影响.结果 54例鼻咽癌组织hMLH1基因启动子甲基化频率为42.59%(23/54),鼻咽癌组织中hMLH1基因启动子甲基化与临床资料无关.23例甲基化鼻咽癌组织hMLH1基因相对转录表达水平为0.939 6±0.037 5,31例非甲基化鼻咽癌组织hMLH1基因相对转录表达水平为0.951 3±0.055 1,两者间差异无统计学意义(P>0.05).结论 鼻咽癌组织中hMLH1基因启动子甲基化与临床资料及转录表达水平均无关,表明其转录表达可能不受甲基化调控.     

大肠癌肿瘤组织中p16基因异常甲基化检测

采用甲基化特异PCR技术 (MSP法 )检测大肠癌患者肿瘤组织中p16基因启动子区异常甲基化改变情况 ,探讨其与大肠癌发生、发展和临床资料的关系。结果显示 ,13例标本 ( 40 .6% )呈p16甲基化阳性 ;DukesC、D期病人肿瘤组织中p16甲基化发生率 ( 63 % )明显高于DukesA、B期病人 ( 2 5 % ) ;在病理分期中 ,高、中分化癌中的p16甲基化阳性率( 3 0 % )低于低分化癌的阳性率 ( 10 0 % ) ;具淋巴结转移癌患者肿瘤组织中p16基因甲基化的阳性率( 63 % )高于淋巴结未转移的阳性率 ( 2 5 % )。研究表明 ,p16甲基化与大肠癌的发生发展和转移呈一定相关性 ,p16甲基化有可能作为大肠癌病人诊断、判断预后的候选标志物之一     

ASPP1基因启动子CpG岛甲基化检测方法的实验初探

目的：建立ASPP1甲基化状况的检测方法,研究ASPP1基因启动子在HEPG-2肝癌细胞株和成纤维细胞株中的甲基化状况。方法：利用因特网对ASPP1基因启动子的CpG岛进行分析,并设计MSP引物,然后对正常的成纤维细胞和p53野生型肿瘤细胞株HEPG-2的ASPP1 DNA启动子甲基化状况进行检测。结果：MSP分析显示,在正常的成纤维细胞中,ASPP1基因启动子CpG岛未出现甲基化;在p53野生型肿瘤细胞株HEPG-2中,ASPP1基因启动子CpG岛处于甲基化状态。结论：MSP检测ASPP1启动子CpG岛甲基化状况的方法客观、准确、可靠。     

焦磷酸测序技术对前列腺癌GSTP1、CDH1、p16基因甲基化的检测研究

目的建立前列腺癌的焦磷酸测序技术为基础DNA甲基化的定量分析方法,并比较良性前列腺增生和前列腺癌组织中GSTP1、CDH1和p16基因DNA甲基化差异,为临床早期诊断前列腺癌奠定基础。方法取前列腺癌组织和良性前列腺增生组织石蜡切片标本,使用焦磷酸测序仪定量检测GSTP1、CDH1和p16基因启动子甲基化状态。结果前列腺癌组织与良性前列腺增生组织比较GSTP1基因超甲基化率为56%(14/25),CDH1基因超甲基化率为32%(8/25),p16基因超甲基化率为20%(5/25)。其中GSTP1、CDH1基因有统计学差异(P<0.05)。结论 GSTP1、CDH1基因在前列腺癌组织中甲基化程度可作为前列腺癌早期诊断的标志物,焦磷酸测序实时定量检测DNA甲基化技术是快速、灵敏、高通量的检测方法。     

肺癌肿瘤抑制因子1在雌激素受体、孕激素受体不同表达乳腺癌患者中的临床意义

目的分析肺癌肿瘤抑制因子1（tumor suppressor in lung cancer 1,TSLC1）与乳腺癌临床病理和预后关系。方法收集女性乳腺癌组织125例,采用免疫组织化学技术检测TSLC1表达。结果TSLC1主要表达在乳腺癌细胞质,其阳性率为38.4%（48/125）,与患者的年龄、肿瘤大小、淋巴结转移、临床分期、人类表皮生长因子受体 2（human epidermal growth factor receptor-2,HER-2）表达无关,而与雌激素受体（estrogen receptor,ER）、孕激素受体（progesterone receptor,PR）表达相关（P<0.001）。ER+（阳性）患者TSLC1阳性率低于ER-（阴性）患者[28.4%（25/88）与62.2%（23/37）];同样,PR+患者TSLC1阳性率低于PR-患者[24.3%（17/70）与56.4%（31/55）]。Z检验显示在ER+PR-的乳腺癌患者中TSLC1-和TSLC1+患者3年生存率差异有统计学意义（83.3%与14.3%,P=0.007）,总体生存曲线差异有统计学意义（P=0.003）。Cox多因素回归分析,TSLC1不是ER+PR-乳腺癌患者独立的危险预后因素（比值比10.696,95%可信区间0.988～115.843,P=0.051）。结论TSLC1在乳腺癌致病机制中与ER、PR相关,可作为临床ER+PR-乳腺癌患者新的预后指标。     

Correlation of apparent diffusion coefficient values measured by diffusion MRI and MGMT promoter methylation semiquantitatively analyzed with MS‐MLPA in patients with glioblastoma multiforme

Purpose:

To retrospectively determine whether the apparent diffusion coefficient (ADC) values correlate with O⁶‐methylguanine DNA methyltransferase (MGMT) promoter methylation semiquantitatively analyzed by methylation‐specific multiplex ligation‐dependent probe amplification (MS‐MLPA) in patients with glioblastoma.

Materials and Methods:

The study was approved by the Institutional Review Board and was Health Insurance Portability and Accountability Act (HIPAA) compliant. Newly diagnosed patients with glioblastoma (n = 26) were analyzed with an ADC histogram approach based on enhancing solid portion. The methylation status of MGMT promoter was assessed by methylation‐specific polymerase chain reaction (MSP) and by MS‐MLPA. MS‐MLPA is a semiquantitative method that determines the methylation ratio. The Ki‐67 labeling index was also analyzed. The mean and 5th percentile ADC values were correlated with MGMT promoter methylation status and Ki‐67 labeling index using a linear regression model. Progression‐free survival (PFS) was also correlated with the ADC values using Kaplan–Meier survival analysis.

Results:

The mean methylation ratio was 0.21 ± 0.20. By MSP, there were 5 methylated and 21 unmethylated tumors. The mean ADC revealed a positive relationship with MGMT promoter methylation ratio (P = 0.015) and was also significantly different according to MSP‐determined methylation status (P = 0.011). Median PFS was significantly related with methylation ratio (P = 0.017) and MSP‐derived methylation status (P = 0.025). A positive relationship was demonstrated between PFS and the mean ADC value (P = 0.001). The 5th percentile ADC values showed a significant negative relationship with Ki‐67 labeling index (P = 0.036).

Conclusion:

We found that ADC values were significantly correlated with PFS as well as with MGMT promoter methylation status. We believe that ADC values may merit further investigation as a noninvasive biomarker for predicting treatment response. J. Magn. Reson. Imaging 2013;37:351–358. © 2012 Wiley Periodicals, Inc.     

基质金属蛋白酶-9在乳腺癌组织中的表达及生物学意义

目的研究基质金属蛋白酶-9（MMP-9）的表达及其与肿瘤临床病理参数之间的关系。方法采用免疫组织化学方法检测109例乳腺癌组织和60例乳腺良性肿瘤组织中的MMP-9的表达并对结果进行分析。结果①109例乳腺癌组织中MMP-9的表达率是40.4%（44/109）,明显高于良性乳腺肿瘤（0.0%）,两者比较差异有统计学意义（P〈0.01）。②MMP-9的表达与病理分级以及临床分期密切相关,随肿瘤直径增大以及肿瘤分期的增加,MMP-9蛋白的阳性表达率也明显增加,差异有统计学意义（P〈0.05）。结论MMP-9蛋白在乳腺癌组织中高表达与乳腺癌局部浸润及预后密切相关,可作为判断乳腺癌预后的重要指标。     

乳腺癌Bcl-2蛋白的表达及临床意义

目的 探讨B细胞淋巴瘤/白血病-2(B cell lymphoma/leukemia-2,Bcl-2)在乳腺癌组织中表达的临床意义.方法 2000年1月-2004年12月就诊的乳腺癌133例,用免疫组织化学法检测Bcl-2蛋白的表达,分析其表达与临床病理特征及预后的关系.结果 Bcl-2在乳腺癌中的表达率为57.9％(...     

早期乳腺癌保乳手术治疗的临床研究

目的探讨早期乳腺癌保乳手术治疗的适应证及方法。方法分析我院近5年来20例保乳手术治疗的乳腺癌患者临床资料。结果保乳术后患者乳房外形均较好,两侧乳房基本对称,患者3年生存率为95％,复发率为5％。结论早期乳腺癌保乳手术治疗安全、疗效确切,全身综合治疗是保乳手术治疗成功的关键。     

Targeted ultrasound for MR-detected lesions in breast cancer patients.

OBJECTIVE: To investigate the usefulness of targeted ultrasound (US) in the identification of additional suspicious lesions found by magnetic resonance (MR) imaging in breast cancer patients and the changes in treatment based on the identification of the lesions by the use of targeted US. MATERIALS AND METHODS: One-hundred forty nine patients who underwent breast MR imaging for a preoperative evaluation of breast cancer between January 2002 and July 2004 were included in the study. We searched all cases for any additional lesions that were found initially by MR imaging and investigated the performance of targeted US in identifying the lesions. We also investigated their pathological outcomes and changes in treatment as a result of lesion identification. RESULTS: Of the 149 patients with breast cancer, additional suspicious lesions were detected with MR imaging in 62 patients (42%). Of the 69 additional lesions found in those 62 patients, 26 (38%) were confirmed as cancers by histology. Thirty-eight lesions in 31 patients were examined with targeted US and were histologically revealed as cancers in 18 (47%), high risk lesions in two (5%), benign lesions in 15 (39%), and unidentified lesions in three (8%). The cancer rate was statistically higher in lesions with a US correlate than in lesions without a US correlate (p = 0.028). Of 31 patients, the surgical plan was altered in 27 (87%). The use of targeted US justified a change in treatment for 22 patients (81%) and misled five patients (19%) into having an unnecessary surgical excision. CONCLUSION: Targeted US can play a useful role in the evaluation of additional suspicious lesions detected by MR imaging in breast cancer patients, but is limited in lesions without a US correlate.     

TIG2基因在肺癌细胞中表达及启动子区甲基化的研究

 目的 探讨TIG2基因在原发性非小细胞肺癌中的表达及意义.方法 应用RT-PCR法检测TIG2基因在肺癌和正常肺细胞中表达变化,用CpG岛预测软件分析TIG2基因启动子和第一外显子区域CpG岛分布,用甲基化特异PCR法(MSP)检测TIG2基因甲基化.用RT-PCR法检测去甲基化试剂作用后TIG2基因的表达.结果 正常肺细胞和癌旁正常组织中TIG2mRNA呈高表达,而肺癌细胞系和组织中低表达或沉默.TIG2基因的启动子和(或)第一外显子区域存在典型的CpG岛.在肺癌细胞系均发生TIG2甲基化,肺癌组织中常发生甲基化,而肺癌旁组织中未见甲基化.去甲基化试剂作用后TIG2基因恢复表达.5-氮-2'-脱氧胞苷浓度越高,TIG2mRNA表达的强度越大.结论 甲基化是导致TIG2基因转录沉默的重要机制,TIG2可能是一种新的肺癌肿瘤抑制基因.     

乳腺癌中MDSCs的扩增和募集及相关临床应用

乳腺癌是女性最常见的恶性肿瘤之一,其转移显著提高了患者的死亡率。肿瘤微环境中多种因素发挥了免疫抑制作用并促进乳腺癌的转移,其中,髓源性抑制细胞（myeloid-derived suppressor cells,MDSCs）是骨髓祖细胞和活化的髓样细胞的异质群体,被认为是肿瘤微环境中发挥免疫抑制作用的重要成分。乳腺癌可以通过多种途径对MDSCs的扩增或募集产生积极影响。作者综述了近年来发现的乳腺癌影响MDSCs扩增或募集的机制,以及这些过程中潜在的治疗靶点。     

Magnetic resonance imaging of breast vascularity in medial versus lateral breast cancer

Objective

Breasts with malignant tumors can demonstrate a general increased vascularity compared to the contralateral breast and a prominent blood vessel adjacent to the tumor on magnetic resonance imaging (MRI). The aim of the study was to further characterize these alterations in blood supply by location of the tumor within the breast using MRI.

Materials and methods

The study group included 105 patients who underwent breast MRI for suspicion of a malignancy over a 2-year period. Fifty-one had pathologically verified malignant tumors (study group), 11 had pathologically verified benign lesions (control), and 43 had negative scans (control). The malignant lesions were distinguished by location, medial or lateral, within the breast. Origin of the vascular supply and vessel diameter was recorded in a blinded manner. When available, MRI scans performed 2 years after treatment were reviewed as well.

Results

Of the 24 medial malignant tumors, 21 (87%) had a predominantly medial vascular supply and 3 (13%), a predominantly lateral supply; of the 23 lateral tumors, 11 (48%) had a predominantly medial vascular supply and 8 (35%), a predominantly lateral supply (p = 0.03). In 4 cases, no dominant vessel was noted. Maximum vessel diameter was 3.6 ± 1.1 mm in the patients with malignancy and 2.6 ± 0.8 mm in the controls (p < 0.0005). General increased vascularity was demonstrated in 91% of the medial tumor subgroup and 83% of the lateral tumor subgroup, as opposed to 36-37% in the control groups (p < 0.0005). Follow-up MRI, performed in 8 patients in the malignant-tumor group after treatment, revealed a considerable decrease in the prominent vessels, to a size close to that of the controls.

Conclusion

Breasts with malignant tumors are characterized by an altered general vascular supply, a prominent feeding vessel, and increased regional vascularity. Both the presence and location of the tumor affect the vascular supply. The vascular change apparently diminishes after treatment, although this finding requires further investigation in a larger sample.     

US correlation for MRI-detected breast lesions in women with familial risk of breast cancer

AIM: To examine the value of US correlation for MRI-detected breast lesions in women with familial risk of breast cancer. METHODS: From an initial dataset of 245 women with positive family history who had breast cancer surveillance involving mammography or MRI between November 1994 and February 2001, 179 subjects with follow-up data were selected. A total of 43 women with 48 MRI-detected lesions underwent further assessment with US. Histopathological correlation was available from 38 breast biopsies performed for 33 women. RESULTS: Sonographic correlates were identified in 32 (66.7%) of the 48 MRI-detected lesions, with cancer present in 11 (34.4%) of these. This compares with 1 (6.3%) cancer found in the 16 lesions without sonographic correlates. Of the 12 malignant lesions, 11 (91.7%) had sonographic correlates whereas 21 (58.3%) of the 36 benign lesions had sonographic correlates. In all 74% of breast biopsies were performed under US guidance compared with 8% under MRI guidance. The proportion of MRI- and US-correlated benign and malignant lesions undergoing US-guided biopsy were 85.7% and 90.9%, respectively. CONCLUSION: The probability of cancer was significantly higher in MRI-detected breast lesions with sonographic correlates compared with those without such correlation. The advantage of convenient biopsy under US guidance as opposed to MRI guidance highlights the value of sonographic assessment of MRI-detected breast lesions.