Comparison of Fluorescence in situ Hybridization and Immunohistochemistry for Assessment of HER-2 Status in Breast Cancer Patients

The accurate assessment ofa proto-oncogene, human epidermal growth factor receptor-2 gene （HER-2）, is extremely important for the therapy and prognosis of breast cancer. Currently, immunohistochemistry （IHC） is the method widely used for the detection of HER-2 protein. Fluorescence in situ hybridization （FISH） has been suggested to be a golden standard assay for HER-2 amplification. This study examined the expression and amplification of HER-2 in paraffin-embedded sections of breast cancer tissues, and compared the two methods on the measurement of HER-2 status. HER-2 gene and protein were determined in breast cancer samples from 52 Chinese women by FISH and IHC respectively. The findings indicated that the HER-2 gene amplification was found in 18 cases （34.6%） by FISH and the HER-2 protein over-expression （score 3＋） in 15 cases （28.8%） by IHC. hnmunohistochemically, 28.6% of the cases scored as 2＋ and 93.3% of the cases scored as 3＋ were HER-2-positive by FISH. There was a significant correlation between the HER-2 gene amplification and HER-2 protein over-expression in breast cancer （P〈0.005）. No correlation was noted between the HER-2 gene amplification and any of the clinicopathological parameters examined, including age, menopausal status, menarche age, tumor size, histological tumor type, histological grade, lymph node status, and the expression of ER and PR. It was concluded that the detection of HER-2 gene amplification in breast cancer by FISH is valuable and can compare with HER-2 protein detection by IHC.

DETECTING EXPRESSION OF MRP-1/CD9 mRNA IN LUNG CANCERS USING TISSUE MICROARRAYS AND FLUORESCENCE IN SITU HYBRIDIZATION METHODS

Objective： The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis and prognosis. Methods： To observe MRP-1/C9mRNA expression, tissue microarray （TMA） containing 54 lung cancers and 10 normal lung tissues was prepared and Fluorescence in situ hybridization was used. Results： The positive rate of MRP-1/CD9 expression was 48.1% in lung cancer, lower than that of normal lung tissues. The statistical difference was significant （P〈0.05）. Its protein expression had no relationship with the patients＇ ages, sex and the macroscopic type of tumor, but had relationships with the histological type, clinical stage, differentiated degree and metastasis. The expression in non-small cell lung cancer （NSCLC） was higher than that in small cell lung cancer （SCLC）; in well-moderately differentiated group was higher than that in poorly differentiated group; Earlier period group （I＋II） was higher than in later period group （Ⅲ＋Ⅳ）; and in group without lymphoid metastasis was higher than in patients with lymphoid metastasis. Conclusion： The progression of the lung cancer maybe related with the descended MRP-1/Cd9 expression, which may be useful in evaluating the prognosis of cancer patients.

Chromosome 11 aneusomy in esophageal cancers and precancerous lesions-an early event in neoplastic transformation:An interphase fluorescence in situ hybridization study from south India

AIM: To detect aneusomic changes with respect to chromosome 11 copy number in esophageal precancers and cancers wherein the generation of cancer-specific phenotypes is believed to be associated with specific chromosomal aneuploidies. METHODS: We performed fluorescence in situ hybridization (FISH) on esophageal tissue paraffin sections to analyze changes in chromosome 11 copy number using apotome-generated images by optical sectioning microscopy. Sections were prepared from esophageal tumor tissue, tissues showing preneoplastic changes and histologically normal tissues (control) obtained from patients referred to the clinic for endoscopic evaluation. RESULTS: Our results demonstrated that aneusomy was seen in all the cancers and preneoplastic tissues, while none of the controls showed aneusomic cells. There was no increase in aneusomy from precancers to cancers. CONCLUSION: Our results suggest that evaluation of chromosome 11 aneusomy in esophageal tissue using FISH with an appropriate signal capture-analysis system, can be used as an ancillary molecular marker predictive of early neoplastic changes. Future studies can be directed towards the genes on chromosome 11,which may play a role in the neoplastic transformation of esophageal precancerous lesions to cancers.

Mesenchymal-epithelial transition factor amplification correlates with adverse pathological features and poor clinical outcome in colorectal cancer

BACKGROUND Colorectal cancer(CRC)is the third most common cancer and the second most common cause of cancer-related mortality worldwide.Mesenchymal-epithelial transition factor(MET)gene participates in multiple tumor biology and shows clinical potential for pharmacological manipulation in tumor treatment.MET amplification has been reported in CRC,but data are very limited.Investigating pathological values of MET in CRC may provide new therapeutic and genetic screening options in future clinical practice.AIM To determine the pathological significance of MET amplification in CRC and to propose a feasible screening strategy.METHODS A number of 205 newly diagnosed CRC patients undergoing surgical resection without any preoperative therapy at Shenzhen Cancer Hospital of Chinese Academy of Medical Sciences were recruited.All patients were without RAS/RAF mutation or microsatellite instability-high.MET amplification and c-MET protein expression were analyzed using fluorescence in situ hybridization(FISH)and immunohistochemistry(IHC),respectively.Correlations between MET aberration and pathological features were detected using the chi-squared test.Progression free survival(PFS)during the two-year follow-up was detected using the Kaplan-Meier method and log rank test.The results of MET FISH and IHC were com pared using one-way ANOVA.RESULTS Polysomy-induced MET amplification was observed in 14.4%of cases,and focal MET amplification was not detected.Polysomy-induced MET amplification was associated with a higher frequency of lymph node metastasis(LNM)(P<0.001)and higher tumor budding grade(P=0.02).In the survival analysis,significant difference was detected between patients with amplified-and non-amplified MET in a two-year follow-up after the first diagnosis(P=0.001).C-MET scores of 0,1+,2+,and 3+were observed in 1.4%,24.9%,54.7%,and 19.0%of tumors,respectively.C-MET overexpression correlated with higher frequency of LNM(P=0.002),but no significant difference of PFS was detected between patients with different protein levels.In terms of concordance between MET FISH and IHC results,MET copy number showed no difference in c-MET IHC 0/1+(3.35±0.18),2+(3.29±0.11)and 3+(3.58±0.22)cohorts,and the MET-to-CEP7 ratio showed no difference in three groups(1.09±0.02,1.10±0.01,and 1.09±0.03).CONCLUSION In CRC,focal MET amplification was a rare event.Polysomy-induced MET amplification correlated with adverse pathological characteristics and poor prognosis.IHC was a poor screening tool for MET amplification.

Comparison of immunohistochemistry with fluorescence in situ hybridization in determining the human epidermal growth factor receptor 2 status of breast cancer specimens： a multicenter study of 3149 Chinese patients

Background Accurate detection of human epidermal growth factor receptor 2 （HER2） expression and gene amplification is crucial for the application of HER2-specific therapy and for evaluating the response of patients with breast cancer.A uniform and standard procedure of immunohistochemistry （IHC） and fluorescence in situ hybridization （FISH） needs to be established for evaluating the HER2 status in breast cancer tissues for the treatment of patients with real HER2-positive tumors.The present multicenter study was aimed to examine the HER2 status in breast cancer specimens from Chinese patients using both IHC and FISH methods.Methods A multicenter study was performed on the HER2 status in 3 149 breast cancer specimens from different ethnic populations and areas in China by IHC and FISH assays.The potential association of HER2 status with demographic and clinical characteristics was analyzed.Results The positive rates for HER2 over-expression and HER2 amplification were 23.3％ and 27.5％ in this study,respectively.The concordance between IHC and FISH was 71.2％ （K=0.494,P ＜0.001）.Furthermore,72.9％ of specimens with IHC 2＋ were negative to FISH.The discordance rates among laboratories were from 5％ to 28％ for IHC and 1％ to 16％ for FISH.HER2 amplification was associated significantly with advanced tumor stage （Ⅲ or Ⅳ,P=0.002）,large tumor size （＞5 cm,P=0.002）,moderate and poor histological grades （P ＜0.0001）,post-menopause （P ＜0.0001）,ER-PR-（P=0.002）,and having ＞4 lymph nodes affected （P ＜0.0001） in this population.The positive rates of HER2 amplification in specimens from Man and Hui Chinese were significantly higher than that in other Chinese populations.There are slightly higher positive rates of HER2 expression and amplification in Chinese patients with breast cancer.Conclusion These findings may provide new insights into understanding the epidemiological features of HER2 expression and amplification,and may be valuable for clinical practice.

TFE3重排肾细胞癌16例临床病理特征及预后分析

目的探讨TFE3重排肾细胞癌(TFE3 rRCC)患者的临床病理特征、免疫组织化学表型、荧光原位杂交、鉴别诊断及预后。方法收集并回顾性分析厦门大学附属第一医院已明确诊断的16例TFE3 rRCC的临床资料、病理学形态特征、免疫组织化学表型、荧光原位杂交的结果。结果患者中女性11例,男性5例,年龄为3~53岁;肿瘤直径3~20 cm。镜下均可见特征性的透明或嗜酸性瘤细胞排列成乳头状结构,伴不同比例的片状、巢状或腺泡样结构。16例TFE3均显示核阳性;13例均存在TFE3基因断裂信号。本组病例随访2~67个月,截止2024年1月仅有1例患者出现脑及新发骨转移病灶,1例患者出现腹部淋巴结转移,其余患者均为无病生存。结论TFE3重排肾细胞癌是一种罕见的恶性肿瘤,免疫组化标记TFE3及荧光原位杂交检测对诊断至关重要,手术为首选治疗,静脉癌栓、AJCC高分期及TFE3均是肿瘤复发的关键因素,目前靶向和免疫治疗尚处于临床试验阶段。

两种国产膀胱癌荧光原位杂交检测试剂盒的临床应用比对

目的:通过比对两种国产膀胱癌荧光原位杂交(Fluorescence In Situ Hybridization,FISH)试剂盒在尿液脱落细胞染色体畸变检测结果差异,探究膀胱癌FISH试剂盒的实际临床应用价值。方法:选取2021年1月至2022年1月,首都医科大学临床检验中心20例血尿患者尿液筛查样本,分别使用不同厂家试剂盒,检测尿液脱落细胞染色体畸变,并与液基薄层细胞特殊染色(CellDetect染色)结果进行对比,分析两种不同试剂盒检测结果的差异。结果:20例样本中,19例样本检测结果一致,仅有1例样本结果不一致。两种试剂盒阳性检出率分别为50%和45%,结果无统计学差异(P>0.05)。结论:两种国产膀胱癌FISH试剂盒检测结果一致性高,均可应用于临床膀胱癌FISH检测。

乳腺癌HER2状态、临床病理特征及预后分析

目的探讨乳腺癌患者的HER2状态、临床病理特征及预后之间的关系。方法选取福州大学附属省立医院629例同时做过HER2 IHC及FISH检测的浸润性乳腺癌手术病例,应用2019版乳腺癌HER2检测指南分析HER2状态、临床病理特征及预后。结果IHC结果:0~1+、2+、3+分别占53.10%、22.26%、24.64%;FISH结果:阴性和阳性病例分别占66.77%和33.23%。导管癌、小叶癌及特殊类型肿瘤之间FISH检测结果差异具有统计学意义(P=0.001),而IHC检测结果差异无统计学意义(P>0.05)。不同性别、发病年龄、肿瘤最大径、组织学分级、淋巴结转移与否两种方法检测结果差异无统计学意义(P>0.05)。针对女性病例,IHC与FISH两种检测方法比较差异具有统计学意义(P<0.001);明确阴性跟阳性病例中,一致性检验,Kappa=0.925,P=0.000;相关性分析,r=0.696,P=0.000。随访病例生存曲线可见赫赛汀治疗组DFS高于非治疗组,IHC阴性FISH阳性病例组有无赫赛汀治疗患者差异具有统计学意义(P<0.05);一致病例与不一致病例DFS比较,差异无统计学意义(P>0.05)。结论基于2019版乳腺癌HER2检测指南,FISH检测结果与肿瘤类型有关;IHC与FISH检测结果具有很好的一致性,二者结果呈正相关;FISH法的检出率高于IHC法,条件允许时,建议做FISH检测确认;结果不一致时,FISH结果对临床制定治疗决策可能更具指导意义。

高通量测序数据再分析在识别结直肠癌患者ERBB2扩增中的价值

目的评估未涵盖即ERBB2拷贝数变异(copynumbervariation,CNV)分析的高通量测序(highthroughput sequencing,HTS)数据再分析在识别结直肠癌患者ERBB2扩增中的价值。方法回顾性分析经病理活检确诊为结直肠癌并接受外周血cfDNA高通量测序样本的检测数据252例。根据经免疫组织化学(immunohistochemistry,IHC)和/或荧光原位杂交(fluorescence in situ hybridization,FISH)判断为ERBB2未扩增样本的HTS数据,计算17号染色体(Chr17)reads数占总reads数的比例范围,初步确定其为识别ERBB2扩增的阈值。根据初步阈值筛选可疑阳性样本,通过数字PCR、IHC和FISH进行验证。结果89例经IHC和/或FISH判定为ERBB2未扩增样本的Chr17 reads数与总reads数的比值为0.239±0.192(0.188~0.299)。以均数的1.25倍0.298作为识别ERBB2扩增的阈值分析其余163例样本数据,发现7例疑似阳性,比值为0.302~0.853。其中5例经IHC和/或FISH判定为阳性,6例经数字PCR验证为阳性,Chr17 reads与总reads数的比值与ERBB2/EIF2C1比值成正相关,相关性好(r2=0.909)。结论再分析未涵盖ERBB2 CNV分析的HTS数据可以初步识别结直肠癌患者的ERBB2扩增。

免疫组化及荧光原位杂交检测人类表皮生长因子受体2的临床意义

目的:探讨结直肠癌中不同免疫组化评分标准下人类表皮生长因子受体2(HER2)蛋白表达情况及荧光原位杂交方法检测HER2基因扩增与临床病理特征之间的关系。方法:选择2019年1月-2021年12月结直肠癌根治术的石蜡组织标本100例,同时收集患者的临床和病理信息。采用三种不同评分标准对所有病例行全自动免疫组化(IHC)检测HER2蛋白和荧光原位杂交(FISH)检测HER2基因表达,分析其与临床病理特征的关系。结果:100例结直肠癌组织中,三种不同评分标准下HER2的蛋白阳性表达率分别为34%,27%和20%,差异无统计学意义(P>0.05)。IHC评分标准1下HER2蛋白表达3+,2+,1+,0+与基因扩增的一致性分别为50.0%,12.5%,2.9%和0。IHC评分标准2下HER2蛋白表达与基因扩增的一致性分别为71.4%,15.0%,2.9%和0,IHC评分标准3下HER2蛋白表达与基因扩增的一致性分别为100.0%,20.0%,2.9%和0,IHC评分标准3下HER2蛋白与基因扩增一致性最高。FISH检测HER2基因扩增9例(9.0%)。HER2基因扩增与年龄、性别、病理分期、浸润深度无相关性(P>0.05),与分化程度、淋巴结转移有相关性(P<0.05)。结论:三种评分标准检测HER2蛋白表达阳性率比较,差异无统计学意义,但评分标准3下HER2蛋白表达与基因扩增的一致性最高。HER2基因扩增与分化程度和淋巴结转移有关。临床上应优先选用评分标准3进行HER2蛋白表达检测。

Association of Fusobacterium nucleatum infection with colorectal cancer in Chinese patients

AIM: To investigate Fusobacterium nucleatum (F. nucleatum) abundance in colorectal cancer (CRC) tissues and its association with CRC invasiveness in Chinese patients.METHODS: The resected cancer and adjacent normal tissues (10 cm beyond cancer margins) from 101 consecutive patients with CRC were collected. Fluorescent quantitative polymerase chain reaction (FQ-PCR) was applied to detect F. nucleatum in CRC and normal tissues. The difference of F. nucleatum abundance between cancer and normal tissues and the relationship of F. nucleatum abundance with clinical variables were evaluated. Fluorescence in situ hybridization (FISH) analysis was performed on 22 CRC tissues with the highest F. nucleatum abundance by FQ-PCR testing to confirm FQ-PCR results.RESULTS: The median abundance of F. nucleatum in CRC tissues [0.242 (0.178-0.276)] was significantly higher than that in normal controls [0.050 (0.023-0.067)] (P < 0.001). F. nucleatum was over-represented in 88/101 (87.1%) CRC samples. The abundance of F. nucleatum determined by 2-ΔCT was significantly greater in tumor samples [0.242 (0.178, 0.276)] than in normal controls [0.050 (0.023, 0.067)] (P < 0.001). The frequency of patients with lymph node metastases was higher in the over-abundance group [52/88 (59.1%)] than in the under-abundance group [0/13 (0%)] (P < 0.005). No significant association of F. nucleatum with other clinico-pathological variables was observed (P > 0.05). FISH analysis also found more F. nucleatum in CRC than in normal tissues (median number 6, 25th 3, 75th 10 vs 2, 25th 1, 75th 5) (P < 0.01).CONCLUSION: F. nucleatum was enriched in CRC tissues and associated with CRC development and metastasis.

EXPRESSION AND SIGNIFICANCE OF SURVIVIN mRNA IN LUNG CANCER TISSUE MICROARRAY DETECTED BY FISH

Objective To investigate the expression of Survivin mRNA in lung cancer tissue microarray （TMA） by fluorescence in .situ hybridization （FISH） method, and determine the role and significance of it in lung cancer genesis and progress. Methods The expression of Survivin mRNA was detected by FISH method and TMA technology. Fifty-four cases of lung cancer and 10 cases of normal lung tissue were examined. Survivin mRNA was expressed in 66.7% （36/54） of lung cancer; the positive ratio of lung cancer was significantly higher than that of normal lung tissue （0/10;X^2= 15.238, P 〈 0.05）. The positive ratio of Survivin mRNA was significantly higher in poor differentiated cancer （20/24, 83.3% ） than moderate and well differentiated cancer （16/30, 53.3%; X^2 = 5.40, P 〈 0.05）. The positive ratio of Survivin mRNA was significantly higher in group with lymph node metastasis （27/32, 84.4%） than without lymph node metastasis （9/22, 40.9%; X^2= 11.084, P 〈 0.05）. The positive ratio of Survivin mRNA was significantly higher in stage Ⅲ-Ⅳ（12/13, 92.3%） than stage Ⅰ- Ⅱ （24/41,58.5%; X^2=5.066, P〈 0.05）. Conclusion Survivin mRNA highly expresses in lung cancer, which is related to the progress and malignant behavior. Survivin may play a promoting role in lung cancer genesis and progress and provide a basis for estimating prognosis and treatment.

Clinicopathologic characteristics and therapeutic responses of Chinese patients with non-small cell lung cancer who harbor an anaplastic lymphoma kinase rearrangement

Introduction: The rearrangement of the anaplastic lymphoma kinase(ALK) gene accounts for approximately 1%–6% of lung adenocarcinoma cases and deines a molecular subgroup of tumors characterized by clinical sensitivity to ALK inhibitors such as crizotinib. This study aimed to identify the relationship between ALK rearrangement and the clinico?pathologic characteristics of non?small cell lung cancer(NSCLC) and to analyze the therapeutic responses of crizotinib and conventional chemotherapy to ALK rearrangement in NSCLC patients.Methods: A total of 487 lung cancer patients who underwent testing for ALK rearrangement in our department were included in this study. ALK rearrangement was examined by using fluorescence in situ hybridization(FISH) assay.Results: Among the 487 patients, 44(9.0%) were diagnosed with ALK rearrangement by using FISH assay. In 123 patients with adenocarcinoma who were non?smokers and of a young age(≤58 years old), the frequency of ALK rearrangement was 20.3%(25/123). Short overall survival(OS) was associated with non?adenocarcinoma tumor type(P = 0.006), poorly diferentiated tumors(P al growth factor rece= 0.001), advanced?stage tumors(P < 0.001), smoking history(P ptor(EGFR)(P = 0.008), and wild?type epidermrter time to cancer p= 0.008). Moreover, patients with poorly diferentiated and advanced?stage tumors had a shorogression compared with those with well diferentiated(P = 0.023) and early?stage tumors(P = 0.001), respectively.Conclusions: ALK?rearranged NSCLC tends to occur in younger individuals who are either non?smokers or light smokers with adenocarcinoma. Patients with ALK rearrangement might beneit from ALK inhibitor therapy.

Correlations of β-catenin,Ki67 and Her-2/neu with gastric cancer

Objective:To study the clinical pathologic characteristics ofβ-catenin,Ki67 and Her-2/neu in gastric cancer and the correlation ofβ-catenin and Ki67 to the protein expression and gene conditions of Her-2/Neu.Methods:The protein expression ofβ-catenin,Ki67 and Her-2/Neu was detected by immunohistochemistry in 101 cases of gastric cancer and the gene conditions of Her-2/Neu by fluorescence in situ hybridization(FISH).Results:The protein expression ofβ-catenin,Ki67 and Her-2/Neu had close relationship with the clinical pathologic characteristics of gastric cancer.Theβ-catenin and Ki67 had obvious correlation to the differentiation,infiltration and lymphatic metastasis of the gastric cancer(P<0.05).The Ki67 had close relationship with the tumor-node-metastasis staging staging of gastric cancer(P<0.05).Her-2/Neu had close relationship with the differentiation and tumor-node-metastasis staging of gastric cancer(P<0.05)but had no relationship with the infiltration and lymphatic metastasis of the gastric cancer(P<0.05).The protein expression of Ki67 had significantly positive correlation to the protein expression and gene amplification conditions of Her-2/Neu(r=0.567,P<0.05 for protein;r=0.304,P<0.05 for gene).Conclusions:Combined detection ofβ-catenin,Ki67 and Her-2/Neu can be used as a reliable method to help the observation of biological behavior,diagnosis and prognosis of gastric cancer,and Ki67 can be used to serve the preliminary screening of Her-2/Neu gene state.

Expression of VEGFR2 and NRP-1 in non-small cell lung cancer and their clinical significance

Objective： Vascular-targeted therapy is gradually becoming more appealing for patients with lung cancer. It is unclear whether vascular endothelial growth factor receptor 2（VEGFR2） and neuropilin-1（NRP-1） can be biomarkers for clinical treatment. We aimed to investigate the expression levels of VEGFR2 and NRP-1 in human non-small cell lung cancer（NSCLC） and their clinical significance by observing patient prognosis. Methods： VEGFR2 and NRP-1 were assessed by immunohistochemistry（IHC） in 40 patients with NSCLC and in 10 patients with benign lesions of lung; kinase insert domain receptor（KDR） and NRP-1 copy number gain（CNG） was assessed by fluorescence in situ hybridization（FISH）. The distributions of overall survival（OS） and progression-free survival（PFS） were estimated using the Kaplan-Meier method and compared between groups by log-rank test.Results： Rates of positive immunostaining for VEGFR2 and NRP-1 were 58% and 55%, respectively. KDR and NRP-1 CNG（＋） were detected in 32.5% and 30% of tumors, respectively. Levels of both VEGFR2 and NRP-1 in lung tumors were significantly different than in the control tissue（χ2=11.22, P=0.001; χ2=9.82, P=0.001, respectively）; similar results were obtained using CNGs（χ2=4.39, P=0.036; χ2=3.95, P=0.046, respectively）. Statistically significant correlations were observed with histological grade, clinical TNM stage and the lymph node status（P〈0.05）, but not age, gender or pathology type（P〉0.05）. VEGFR2 showed a strong correlation with NRP-1（Rs=0.68, P=0.00）; similar results were observed with KDR and NRP-1 CNG（Rs=0.32, P=0.04）. Significant differences in OS and PFS were observed between the groups with higher VEGFR2 and NRP-1 and those with lower expression（P〈0.05）. Conclusions： According to these data, VEGFR2 and NRP-1 are highly expressed in NSCLC. We can conclude that they play a key role in NSCLC occurrence, development and metastasis and are associated with patient prognosis（P〈0.05 for OS and PFS）. This information will be beneficial for clinical antiangiogenic treatment in NSCLC.

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.

Detection of TMPRSS2：ERG fusion gene in circulating prostate cancer cells

Aim： To investigate the existence of TMPRSS2：ERG fusion gene in circulating tumor cells （CTC） from prostate cancer patients and its potential in monitoring tumor metastasis. Methods- We analyzed the frequency of TMPRSS2： ERG and TMPRSS2：ETV1 transcripts in 27 prostate cancer biopsies from prostatectomies, and TMPRSS2：ERG transcripts in CTC isolated from 15 patients with advanced androgen independent disease using reverse transcription polymerase chain reaction （RT-PCR）. Fluorescence in situ hybridization （FISH） was applied to analyze the genomic truncation of ERG, which is the result of TMPRSS2：ERG fusion in 10 of the 15 CTC samples. Results： TMPRSS2： ERG transcripts were found in 44% of our samples, but we did not detect expression of TMPRSS2：ETV1. Using FISH analysis we detected chromosomal rearrangements affecting the ERG gene in 6 of 10 CTC samples, including 1 case with associated TMPRSS2：ERG fusion at the primary site. However, TMPRSS2：ERG transcripts were not detected in any of the 15 CTC samples, including the 10 cases analyzed by FISH. Conclusion： Although further study is required to address the association between TMPRSS2：ERG fusion and prostate cancer metastasis, detection of genomic truncation of the ERG gene by FISH analysis could be useful for monitoring the appearance of CTC and the potential for prostate cancer metastasis.

Cervical cancer screening: hTERC gene amplification detection by FISH in comparison with conventional methods

Aim: To assess the clinical significance of hTERC amplification for cervical cancer screening detected by fluorescence in situ hybridization (FISH) and compare it with that of current screening methods within the same group. Methods: A total of one hundred and nine women were recruited in this study. All of them had liquid-based thin-prep cytologic test (TCT), human papillomavirus (HPV) DNA testing and hTERC gene amplification analysis using interphase two-color FISH. In addition, colposcopically directed biopsy and/or cone biopsy were conducted for definite histopathologic diagnosis for each case. The optimal threashold of hTERC gene amplification by fluorescence in situ hybridization (FISH) were assecced by receiver operating characteristic (ROC) curve. The results of hTERC gene amplification analysis were compared with the cytological analysis, HPV DNA testing and those of subsequent biopsies. Results: Among the 109 patients, 18 were benign lesion, 17 were LSIL, 66 were HSIL and 8 were invasive carcinoma of cervix (ICC). Of them, hTERC-positive cases were found in 0.0% (0/18) of normal specimens, 11.8% (2/17) of LSIL, 72.7% (48/66) of HSIL and 100.0% (8/8) of ICC, respectively. The positive rate of hTERC gene amplification was significantly higher in HSIL and ICC compared with normal and LSIL (all P < 0.01).The optimal cut-off point of percentages of cells with hTERC amplification was determined as 5.5%. Using this threshold the hTERC test reached a much higher specificity(94.3%, 33/35) and a relatively lower sensitivity(77.0%, 57/74) to distinguish benign lesion and LSIL from HSIL and ICC in comparison with HR-HPV test (51.4%;91.9%) and TCT (74.3%;81.1%). Area Under the Curve revealed that hTERC amplification test performed more accurately (area under the curve = 0.857) compared to HPV test (area under the curve = 0.717) and cytology(area under the curve = 0.777) to discriminate HSIL or higher from LSIL or lower. This study also found a significant positive correlation between positive hTERC gain and HR-HPV infection, abnormal cytological or histopathologic lesions (all P < 0.01) in patients with cervical diseases. Conclusion: hTERC amplification testing may be a promising adjunct to screen women for cervical precancer or cancer with high specificity and accuracy.

Predictive biomarker and clinicopathological characteristics analysis for recurrence of early gastric cancer

Objective: The aim of this study was to identify the correlation between the clinicopathological characteristics and recurrence in early gastric cancer (EGC), what's more, we attempt to look for a predictive biomarker to predict and treat for re-currence of EGC. Methods: This study retrospectively analyzed 178 early gastric cancer patients who had the complete post-operative and follow-up medical records in the First Affiliated Hospital of Yangtze University (China) between January 1995 to December 2005. All of them were followed-up to December 2009 regularly. Computer tomography (CT), endoscopy, and single photon emission computed tomography (SPET-CT) were used to diagnose for recurrence of EGC. Immunohistochem-istry (IHC) and fluorescence in situ hybridization (FISH) were used for the detection of cerbB2. Chi-square test was applied to this study for statistics analysis. Results: Fourteen patients had recurrence. Eighteen patients were cerbB2-positive, including twelve recurrence patients and six norecurrence patients. Sex, tumor depth, and lymph node metastasis were related to the recurrence of EGC. Also, cerbB2-positive patients had the higher recurrence rate compared to the cerbB2-negative patients. Conclusion: Recurrence of EGC after curative resection can be predicted by using some clinicopathological characteristics. CerbB2 can be used as a predictive biomarker for recurrence of EGC.

人表皮生长因子受体2在结直肠癌中的表达及其临床意义

目的探讨人表皮生长因子受体2(HER-2)在结直肠癌中的表达及其与临床病理特征的关系。方法选取2019年4月至2022年1月徐州市肿瘤医院病理科100例结直肠癌标本设为结直肠癌组,将其中距肿瘤5 cm以上的60例正常肠黏膜设为正常肠黏膜组,另选取同期的60例结直肠腺瘤标本设为结直肠腺瘤组。三组均采用免疫组织化学(IHC)方法和荧光原位杂交(FISH)技术检测。分析HER-2在三组中的表达情况,比较三组HER-2(2+)基因扩增结果、三组HER-2的表达、结直肠癌不同临床病理特征的HER-2表达。结果结直肠癌组中,HER-2(3+)15例,HER-2(2+)45例,HER-2(0)或(1+)40例。结直肠腺瘤组中,HER-2(3+)7例,HER-2(2+)22例,HER-2(0)或(1+)31例。正常肠黏膜组中HER-2(2+)7例,HER-2(0)或(1+)53例。三组的HER-2(2+)基因扩增结果比较,差异无统计学意义(P>0.05)。结直肠癌组、结直肠腺瘤组的HER-2阳性表达均显著高于正常黏膜组,差异有统计学意义(P<0.05);结直肠癌组与结直肠腺瘤组的HER-2阳性表达比较,差异无统计学意义(P>0.05)。不同性别、年龄、肿瘤直径、TNM分期的HER-2阳性表达比较,差异无统计学意义(P>0.05);“高+中分化”的HER-2阳性表达低于“低分化”,“Tis+T1+T2”浸润深度的HER-2阳性表达低于“T3+T4”浸润深度,有淋巴结转移的HER-2阳性表达高于无淋巴结转移,差异有统计学意义(P<0.05)。结论HER-2在结直肠癌和腺瘤中均存在高表达,与癌组织的分化程度、浸润深度及淋巴结转移有密切关系。