Dual fluorescence in situ hybridization in detection of HER-2 oncogene amplification in primary hepatocellular carcinoma

BACKGROUND: Molecular cytogenetics of oncogene HER-2 amplification in primary hepatocellular carcinoma (HCC) is still unknown. The aim of this study was to in vestigate the frequency of HER-2 oncogene amplification in primary HCC and its relations to clinicopathological pa rameters and prognosis. METHODS: Forty-two surgical samples from patients with primary HCC were detected for their HER-2 oncogene am plification. The number of chromosome 17 and their ratio were tested by dual fluorescence in situ hybridization (FISH) technique, and then the correlations between HER-2 amplification, clinicopathological characteristics and prog nosis were analyzed statistically. RESULTS: HER-2 oncogene amplification was detected in 9 (21.4%) of the 42 primary HCCs, including 4 patient with high copy (HC) (9.5%) and 5 patients with low copy (LC) (11.9%). HER-2 amplification was associated signifi cantly with tumor size and postoperative survival time o HCC patients (P<0.05), and the presence of HER-2 gene amplification was correlated with postoperative relapse (P— 0.257), but not related to sex, age, AFP level, HBV infec tion, histopathological grading and clinical staging of HCC patients (P>0.05). The HER-2 oncogene copy was exa mined in 31 (73.8%) of the 42 primary HCCs, consisting of 9 patients with HER-2 amplification (21.4%) and 22 pa tients with aneuploidy (52.4%). No significant relation were observed between the HER-2 oncogene copy, patien sex, tumor size, histopathological grading, clinical stag ing, postoperative relapse and survival time (P >0.05); bu the HER-2 oncogene copy was correlated significantly to age, AFP level and HBV infection (P <0.05). CONCLUSIONS: There are a lower frequency of HER-2 oncogene amplification and a higher frequency of chromo- some 17 aneuploidy in primary HCC. HER-2 oncogene amplification may be involved in the development and pro- gression of large HCC in some patients, and seems to be a valuably independent prognostic factor predicting the re- currence and poor survival in patients with large HCC.

Study of differential polymerase chain reaction of C-erbB-2 oncogene amplification in gastric cancer

AIM To study the significance of C-erbB-2 oncogene amplification in gastric cancer.METHODS C-erbB-2 oncogene amplification was examined by using differential polymerase chain reaction (dPCR) in surgical and endoscopic specimens of 83 cases of gastric cancer and 101 metastatic lymph nodes.RESULTS C-erbB-2 amplification was found in 28.9% (24/ 83) surgical specimens and 20.5% (17/ 83) endoscopic ones of gastric cancer patients. The amplification was significant in both types of specimens of advanced cancer cases (P＜0.05) and surgical specimens with lymph node metastasis (P＜0.01). The incidence of C-erbB-2 amplification in lymph nodes with metastasis was higher than in primary sites (surgical specimens, P＜0.05). The patients with amplification tumors had poorer 5-year survival rates than those with unamplification ones in the early cancers and well to moderately differentiated adenocarcinomas (P＜0.05). The same surgical samples were tested again by Southern blot hybridization to ascertain C-erbB-2 amplification, and the positive rate of C-erbB-2 amplification (15.7%) was lower than that of dPCR (28.9%, P＜0.05).CONCLUSION Examining C-erbB-2 amplification by dPCR is a quick, simple, reliable and independent method, and is helpful in predicting prognosis and metastatic potential of gastric cancer.

The expression of HER-2/neu gene in colon cancer tissues and its clinical significance

Objective：The article aims to detect the expression of HER-2/neu gene in colon cancer tissues and adjacent tissues, to analyze the relationship between dif erent pathologic types and clinical features, also to invest the distribution of patients with positive expression of HER-2 gene. Methods：The expression of HER-2 gene in the 223 samples with colon can-cer was detected by immunochemical approach. The expression of HER-2 gene in colon cancer tissues and adjacent tissues and dif erent pathologic types was analyzed byχ2 test. The correlation between the expression of HER-2 gene and clinical features was analyzed by Spearman. Results：The number of positive expression of HER-2 gene in colon cancer tissues and adjacent tissues were 74 and 0 respectively, the dif erence has statistical significance. The number of papil ary or tubular adenocarcinoma was 182, among them, 60 cases were positive expression. The number of mucinous adenocarcinoma was 41, among them, 14 cases were positive expression. The expression of HER-2/neu gene has no correlation with sex, age, the maximum diameter, general classification, degree of dif erentiation and depth of invasion, which has no statistical significance. However, the expression of HER-2/neu gene has correlation with metastasis of lymph node and Dukes stage, which has statistical significance. The expression of HER-2/neu gene was positive correlation with metastasis of lymph node and Dukes stage. The correlated coef icient index was 0.320 and 0.320 respectively. In the 74 patients with positive expression of HER-2 gene, 59.4%of them were 60-74 years old. And there was 97.3%of the patients without family history of adenocarcinoma. Conclusion：The expression of HER-2/neu gene in colon cancer tissues was higher than in adjacent tissues. The expression of HER-2/neu gene has no correlation with sex, age, the maximum diameter, general classification, degree of dif erentiation and depth of invasion, but has correlation with metastasis of lymph node and Dukes stage. The expression of HER-2/neu gene was positive correlation with metastasis of lymph node and Dukes stage. The expression of HER-2/neu gene with age of 60-74 years old and without family history of adenocarcinoma was higher than other groups.

荧光原位杂交检测胃癌中HER-2基因状态

目的探讨胃癌中HER-2基因扩增与HER-2蛋白过表达的关系。方法运用免疫组化（IHC）和荧光原位杂交（FISH）技术,对手术切除的84例胃癌石蜡标本进行HER-2状态的检测。结果84例中,19例HER-2蛋白（＋）（22.6%）,其中HER-2基因扩增7例（8.3%） 12例HER-2蛋白（/）（14.3%）中,HER-2基因扩增5例（6%）,占高表达组的41.7% 72例HER-2蛋白（＋/0）（85.8%）中,HER-2基因扩增2例（2.4%）。结论HER-2蛋白（/）与HER-2基因扩增密切相关,故这类患者可考虑先做IHC初步筛选,再做FISH确诊,从而为胃癌分子靶向治疗提供有价值的信息。

乳腺癌HER-2基因荧光原位杂交检测的临床应用

目的研究荧光原位杂交(FISH)用于乳腺癌HER-2检测的临床应用及HER-2基因扩增与乳腺癌临床病理的相关性。方法应用FISH技术和免疫组化(IHC)技术检测40例乳腺浸润性导管癌石蜡包埋标本,对比分析。结果IHC检测CerbB-2(3+)/(2+)/(1+或0),其FISH阳性率分别为85.7%、50%、5.9%。24例腋窝淋巴结阳性者,其FISH检测HER-2基因扩增10例(P=0.0399)。ER/PR阴性8例,其HER-2基因扩增6例(P>0.05)。结论IHC检测HER-2蛋白有很高的假阳性及假阴性,FISH有效性及准确性显著高于IHC,可在临床广泛推广应用。HER-2基因扩增与腋窝淋巴结阳性相关。

胃癌HER-2基因检测方法及其靶向治疗的研究现状

HER-2/neu基因,也称CerbB-2,为人类表皮生长因子受体(EGFR)家族的第2个成员.许多研究认为胃癌组织中存在HER-2基因扩增和/或蛋白的过表达.针对HER-2基因的免疫治疗可能成为晚期胃癌患者新的重要治疗选择,因此,胃癌组织中HER-2基因状态的检测就显得非常重要.本文就胃癌HER-2基因检测方法及其靶向治疗的研究现状进行综述.

乳腺癌HER-2基因扩增与临床病理特征的相关性分析

目的检测乳腺癌患者HER-2基因,并分析其与临床病理特征的相关性。方法采用荧光原位杂交方法(fluorescence in situ hybridization,FISH)检测328例乳腺癌石蜡标本的HER-2基因扩增状况,分析其与临床病理特征之间的关系。结果 328例乳腺癌患者HER-2基因,阳性率26.2%(86/328)。FISH与IHC结果的总符合率为72.0%。HER-2基因扩增状况与年龄差异不具有统计学意义(P>0.05),与肿瘤直径、病理类型、腋淋巴结转移数、临床分级的差异和HER-2受体蛋白表达的差异具有统计学意义(P<0.05)。结论 HER-2基因可作为判断乳腺癌预后及拟订治疗方案的良好指标。

鼻咽癌患者HER-2/neu基因扩增和表达及其临床意义

目的 :研究鼻咽癌HER 2 /neu基因扩增、表达及其临床意义。方法 :采用原位荧光杂交、逆转录多聚链式反应和免疫组化技术检测鼻咽癌组织HER 2 /neu基因扩增、表达。结果 :HER 2 /neu基因在鼻咽癌中无扩增 ,但有过表达 ,其原因是由于mRNA高表达所致 ;这种过表达与鼻咽癌预后之间未显示有相关性。结论 :HER 2 /neu基因在鼻咽癌无扩增 ,有过表达 ,这种过表达未显示预后意义。

乳腺癌手术标本BRCA-1蛋白表达与HER-2基因扩增的临床意义

目的:研究乳腺癌患者乳腺癌手术标本中乳腺癌易感基因1(breast cancer susceptibility gene1,BRCA-1)蛋白表达、原癌基因HER-2的基因扩增与蛋白表达及其与临床病理特征之间的关系。方法:采用免疫组织化学和显色原位杂交的方法,检测乳腺癌手术切除肿瘤组织中的HER-2和BRCA-1蛋白表达以及HER-2基因扩增情况,对这两个基因的表达与临床相关病理特征之间的关系进行分析。结果:BRCA-1和HER-2蛋白表达在不同组织类型中差异无统计学意义,但在不同肿瘤大小、不同组织分级和有无淋巴结转移中差异有显著性(P<0.05)。BRCA-1蛋白表达与HER-2基因扩增率呈负相关。结论:BRCA-1和HER-2蛋白表达对乳腺癌的发生发展具有一定作用,检测BRCA-1和HER-2蛋白表达在乳腺癌的早期诊断、判断生物学行为、预测预后中有重要意义。另外BRCA-1与HER-2可能存在相关性,须进一步研究。

胃癌伴神经内分泌分化HER-2基因扩增和蛋白表达及预后分析

目的观察胃癌伴神经内分泌分化(NED)和胃癌中HER-2基因扩增及蛋白表达情况。方法回顾性分析胃癌伴神经内分泌分化患者70例和具有相同分期的胃癌患者150例,采用免疫组化法(IHC)联合双色银染色素原位杂交(DISH)检测胃癌伴NED和胃癌中HER-2基因扩增和蛋白表达情况,并对胃癌伴NED进行预后分析。结果胃癌伴NED和胃癌中HER-2蛋白过表达率分别为20.00%和21.33%,HER-2基因扩增率分别为8.57%和14.67%。HER-2蛋白表达、HER-2基因扩增及基因扩增类型2组间差异均无统计学意义;胃癌伴NED17号染色体多体与HER-2基因扩增呈正相关。胃癌伴NED患者中HER-2基因扩增者术后生存期短于HER基因不扩增者。多因素分析显示胃癌伴或不伴NED、HER-2基因是否扩增、淋巴结转移和手术方式是预后的独立影响因素。结论胃癌伴NED属于胃癌特殊类型,其HER-2基因扩增、蛋白表达情况和胃癌相同,HER-2基因扩增的胃癌伴NED患者预后差。

Relationship between HER-2 overexpression and brain metastasis in esophageal cancer patients

AIM:To study if HER-2 overexpression by locally advanced esophageal cancers increase the chance of brain metastasis following esophagectomy.METHODS:We retrospectively reviewed the medical records of esophageal cancer patients who underwent esophagectomy at University of Iowa Hospitals and Clinics between 2000 and 2010.Data analyzed consisted of demographic and clinical variables.The brain metastasis tissue was assayed for HER-2 overexpression utilizing the FDA approved DAKO Hercept Test.RESULTS:One hundred and forty two patients were reviewed.Median age was 64 years(36-86 years).Eighty eight patients(62%) received neoadjuvant chemoradiotherapy.Pathological complete and partial responses were achieved in 17(19%) and 71(81%) patients.Cancer relapsed in 43/142(30%) patients.The brain was the first site of relapse in 9/43 patients(21%,95% CI:10%-36%).HER-2 immunohistochemistry testing of the brain metastasis tissue showed that 5/9(56%) cases overexpressed HER-2(3+ staining).CONCLUSION:HER-2 overexpression might be associated with increased risk of brain metastasis in esophageal cancer patients following esophagectomy.Further studies will be required to validate this observation.

HER-2 expression after neoadjuvant chemotherapy of the breast cancers

Objective:The aim of this study was to study changes of HER-2 expression after neoadjuvant chemotherapy in the breast cancer cases.Methods:One hundred and thirty-seven female patients with primary breast cancers,who received neoadjuvant chemotherapy,underwent core needle puncture and Mammotome biopsy before chemotherapy,and the biopsy results were used as the basis of histological diagnosis,fluorescence in situ hybridization (FISH) was performed to test HER2 status of tumor tissues before and after chemotherapy.All patients underwent FEC,TE,or AC neoadjuvant chemotherapy of 2-6 cycles before surgery.Results:Twenty-two patients were positive according to FISH test among 137 preoperative patients,8 patients achieved pathological complete remission after chemotherapy (three HER-2 positive patients and five negative patients),91 patients achieved partial remission,24 patients were stable,and 14 cases were invalid.Twenty-two patients were positive according to FISH test (8 patients with pathological complete remission did not undergo test),and positive patients still expressed positively after chemotherapy before neoadjuvant chemotherapy.Three negative patients were converted to be positive,and changes before and after chemotherapy had no statistical difference (P>0.05).Conclusion:Neoadjuvant chemotherapy makes no influence on patients with HER-2 positive expression,while patients with negative expression can be converted to be positive,but without significant difference.

NDRG2 gene copy number is not altered in colorectal carcinoma

AIM To investigate if the down-regulation of N-myc Downstream Regulated Gene 2(NDRG2) expression in colorectal carcinoma(CRC) is due to loss of the NDRG2 allele(s).METHODS The following were investigated in the human colorectal cancer cell lines DLD-1, Lo Vo and SW-480: NDRG2 mRNA expression levels using quantitative reverse transcriptionpolymerase chain reaction(qRT-PCR); interaction of the MYC gene-regulatory protein with the NDRG2 promoter using chromatin immunoprecipitation; and NDRG2 promoter methylation using bisulfite sequencing.Furthermore, we performed qPCR to analyse the copy numbers of NDRG2 and MYC genes in the above three cell lines, 8 normal colorectal tissue samples and 40 CRC tissue samples.RESULTS As expected, NDRG2 mRNA levels were low in the three colorectal cancer cell lines, compared to normal colon.Endogenous MYC protein interacted with the NDRG2 core promoter in all three cell lines.In addition, the NDRG2 promoter was heavily methylated in these cell lines, suggesting an epigenetic regulatory mechanism.Unaltered gene copy numbers of NDRG2 were observed in the three cell lines.In the colorectal tissues, one normal and three CRC samples showed partial or complete loss of one NDRG2 allele.In contrast, the MYC gene was amplified in one cell line and in more than 40% of the CRC cases.CONCLUSION Our study suggests that the reduction in NDRG2 expression observed in CRC is due to transcriptional repression by MYC and promoter methylation, and is not due to allelic loss.

EXPRESSION OF MOUSE Tbx2 GENE IN NORMAL AND MALIGNANT MELANOPHORES BY RT-PCR

Objective: To observe the expression of mouse Tbx2 gene in normal and malignant melanophore. Methods: The normal and malignant cells were used to extract total RNA. The expression of the Tbx2 gene was detected by RT-PCR. Results: No expression of the Tbx2 gene in the normal melanocytes was noted, but all malignant cells showed expression of the Tbx2 gene. Conclusion: Tbx2 plays a critical role during the development of the malignant cells.

c-erbB-2基因生物学特性研究进展

c-erbB-2是一种细胞来源的原癌基因,编码具有蛋白激酶活性的细胞膜糖蛋白。本文从c-erbB-2基因的生物学活性,激活作用机制以及对该基因扩增引起各种肿瘤的发生做了综述,并探讨了该基因在肿瘤诊断预测中的重要作用和应用前景。

玉米果糖-6-磷酸,2-激酶/果糖-2,6-二磷酸酶基因的全长cDNA克隆及表达

以来自“掖单 4号”的玉米果糖 6 磷酸 ,2 激酶 果糖 2 ,6 二磷酸酶 (F2KP)基因cDNA片段 (AF0 0 75 82 )为基础 ,运用RT PCR和RACE技术 ,从“紫玉糯 1号”中获得了 1个 2 4 6 9bp的玉米F2KP基因cDNA克隆 ,命名为mF2KP ,GenBank登录号为AF33414 3。该cDNA包含 1个 2 2 2 6bp的开放阅读框 ,编码 74 1个氨基酸。序列分析表明 ,两个玉米品种的F2KP基因存在一定差异 :mF2KP基因的 3′端非编码区比AF0 0 75 82序列短 38bp ;在mF2KP的15 92、15 93和 16 0 5位置上 ,分别比AF0 0 75 82序列多出了 1个碱基 ,导致阅读框在一个小范围内发生了移位。North ern杂交表明 :不同玉米组织中mF2KP的表达差异明显。在茎中mF2KP的表达水平比叶片、苞叶以及雄花序中的表达水平低 ,但比未成熟种子中的表达水平高。在未成熟种子中 。

FISH检测乳腺癌HER2基因扩增及临床病理关系的研究

目的:探讨荧光原位杂交(FISH)检测原发性乳腺癌组织中HER2基因扩增与临床病理的关系。方法:收集2007年11月至2009年8月手术切除的乳腺癌组织标本105例,采用FISH方法检测HER2基因扩增情况,分析其与临床病理的关系,同时比较FISH与免疫组化(IHC)检测结果的相关性。结果:105例乳腺癌中共有39例HER2基因扩增,阳性率为37.1%(39/105),其扩增与HER2蛋白过表达、组织学分级、雌激素和孕激素受体(均P<0.05)状态有关;FISH与IHC两种方法检测HER2结果密切相关(rs=0.680,P<0.05)。结论:HER2基因扩增与乳腺癌发生发展及临床预后密切相关,可作为乳腺癌治疗的理想靶点。

快速检测志贺毒素2基因的LAMP方法的建立

志贺毒素（Shigatoxin，Stx）又称Vero毒素，是产志贺毒素大肠杆菌（Shiga toxin-producing Ecoli，STEC）重要毒力因子之一，具有细胞毒性、肠毒性、神经毒性作用，是导致被感染者出现严重症状甚至死亡的主要原因，因此，有必要建立敏感而快速的检测Stx方法。细菌培养方法需要耗费3d，而快速检测方法如ELISA需要高浓度的病原菌才能检测到。

植物乳杆菌HE-1由AI-2/LuxS介导的群体感应与生物膜形成关系的研究

本研究从AI-2/LuxS群体感应出发,研究植物乳杆菌HE-1的AI-2信号分子生成与生物膜形成的关系,同时对luxS基因进行扩增,从分子生物学角度验证菌株HE-1的AI-2信号分子产生。研究使用生物发光的方法检测植物乳杆菌HE-1不同培养时间AI-2的产生量;使用微孔板法确定不同时间点生物膜的生成量;之后使用PCR的方法扩增其luxS基因片段,对扩增片段进行同源性和系统发育分析。结果显示植物乳杆菌HE-1的生物膜生成时间和生成量与AI-2信号分子的产生时间和产生量非常吻合,luxS基因成功扩增,同源性分析显示植物乳杆菌HE-1与已知植物乳杆菌luxS基因同源性高达99%以上。分析实验结果得出植物乳杆菌HE-1生物膜生成与AI-2/LuxS群体感应系统二者之间有密切的关系,并且luxS基因的扩增成功和同源性分析从分子生物学上验证了植物乳杆菌HE-1具有产生AI-2信号分子的关键基因。

弥漫性大B细胞性淋巴瘤中bcl-2表达与基因重排的相关性

目的探讨弥漫性大B细胞性淋巴瘤(DLBCL)中bcl-2蛋白表达与基因异常间的相关性。方法采用免疫组化方法检测42例不同器官和部位发生的DLBCL石蜡包埋组织中bcl-2蛋白表达,采用双色FISH法检测42例DLBCL石蜡包埋组织中BCL-2基因重排及扩增。观察组织标本中bcl-2蛋白表达与基因异常之间的相关性。结果 BCL-2基因重排阳性率为42.9%(18/42),16例生发中心型(GCB型)阳性率为43.8%(7/16),26例non-GCB型阳性率为42.3%(11/26),统计学分析χ~2=0.008(P>0.05),差别不显著;bcl-2蛋白表达与基因重排列联系数为0.333(P<0.05),差异显著;bcl-2蛋白表达与BCL-2扩增列联系数为0.243(P>0.05),差异不显著。结论 bcl-2蛋白高表达(中强以上,≥50%)与BCL-2基因重排有密切相关性。DLBCL中,bcl-2蛋白高表达提示可能存在BCL-2基因重排。