CA 19-9, CA 125 and CEA in the endometrial mucosa during the menstrual cycle, in atypical hyperplasia and endometrial carcinoma

CA 19-9, CA 125 and CEA were demonstrated by immunohistochemistry in 58 tissue samples of normal mucosa, 21 samples of atypical hyperplasia and 74 samples of endometrial carcinoma. CA 19-9 was mainly detected in the mid phase of secretion (8/11). CA 125 in the mid (6/11) and in the late phase (8/9). As opposed to CEA, both tumor markers are secretion products of the normal endometrium and are not expressed in the endometrial glands during the proliferation phase. CA 125 expression does not correlate with the degrees of differentiation or malignancy. The percentage of CA 19-9 positive cases rises with increasing differentiation. In atypical hyperplasia, however, this percentage is as small as in undifferentiated carcinoma. 93% of the endometrial carcinomas were CA 19-9, 65% CA 125 and 58% CEA positive.     

BAT-26 microsatellite instability does not correlate with the loss of hMLH1 and hMSH2 protein expression in sporadic endometrial cancers

Microsatellite instability (MSI) is detected in about 20-25% of endometrial cancers (ECs). Incidence of this alteration correlates with lack of expression of certain mismatch repair genes such as hMLH1 and hMSH2. Although assessment of several markers has been proposed for identification of microsatellite unstable tumours, BAT-26, a mononucleotide microsatellite repeat, has been shown to be highly efficient when used as a single marker. The aim of the study was to evaluate instability within BAT-26 and expression of hMLH1 and hMSH2 proteins in sporadic endometrial cancer as well as to correlate these findings with histopathologic and clinical characteristics of tumours. Samples of 88 (74 endometrioid and 14 non-endometrioid) ECs were investigated for instability within BAT-26 by means of PCR and expression of hMLH1 and hMSH2 proteins using immunohistochemistry. BAT-26 MIS was discovered in 23.9% of endometrial cancers. Incidence of MSI did not correlated with grade, stage or depth of invasion. BAT-26 MSI was more frequent in non-endometrioid compared to endometrioid tumours (35.7% vs. 21.6%, respectively), but the difference was not statistically significant. Lack of hMLH1 and hMSH2 protein expression was detected in 21.6 and 15.9% of ECs, respectively, and did not correlate with clinicopathologic features of tumours. Loss of both hMLH1 and hMSH2 protein expression was similar in BAT-26 stable and unstable cancers. All cases of non-endometrioid tumours with BAT-26 MSI were positive for hMLH1. We can conclude that BAT-26 used alone may not be a reliable marker for identification of sporadic ECs with microsatellite instability induced by deficient expression of hMLH1 and hMSH2.     

Epidermal growth factor receptor and c-erbB-2 expression in normal breast tissue during the menstrual cycle

Summary EGF receptor (EGF-R) and c-erbB-2 are homologous tyrosine kinase transmembrane receptors. They are involved in controlling proliferation, and probably differentiation, of normal breast epithelial cells, and their expression has been linked to the prognosis of breast cancer. Their physiological roles in normal breast tissue remain to be elucidated, as most studies to date have involved breast cancer cell lines. We studied the location of EGF-R and c-erbB-2 in 100 samples of normal breast with standard immunohistochemical methods and double-labelling techniques. EGF-R was mainly expressed on the stroma and myoepithelial cells, whereas c-erbB-2 expression was exclusively epithelial. An image analyser was used to quantitate variations in their expression during the mentrual cycle. EGF-R and c-erbB-2 expression on epithelial cells was stronger during the luteal phase than the follicular phase (p < 0.01 for EGF-R). The pattern of expression was also compared with that in 28 breast cancers and 7 fibroadenomas.     

Loss of hMSH2 and hMSH6 expression is frequent in sporadic endometrial carcinomas with microsatellite instability: a population-based study.

Microsatellite instability (MSI) seems to be important in the development of various human cancers including sporadic endometrial cancer. It has previously been shown that alterations in the mismatch repair gene hMLH1 seem to be important for the development of MSI in these tumors. The role of the other mismatch repair genes hMSH2 and hMSH6 has been less well studied, but investigations on patients with hereditary nonpolyposis colorectal cancer indicate that these genes also may be involved. We therefore wanted to investigate the pattern of hMSH2 and hMSH6 expression in a prospective and population-based series of endometrial carcinomas with known hMLH1 expression and MSI status. A total of 138 patients were studied, and pathological staining was seen in 19 cases (14%) for hMLH1, 26 cases (19%) for hMSH2, and 17 cases (12.3%) for hMSH6. Pathological hMLH1 expression was more frequent among tumors with high MSI (those positive for four to five of five markers), whereas pathological expression of hMSH2 and hMSH6 was more frequent among tumors with intermediate MSI (those positive for two to three of five markers). MSI was significantly correlated with pathological expression of hMLH1 (P < 0.001), hMSH2 (P = 0.04), and hMSH6 (P = 0.001). In the group with high MSI, 14 of 16 tumors (88%) showed pathological expression for at least one of the markers. The expression of hMLH1, hMSH2, or hMSH6 did not significantly influence survival. In conclusion, pathological expression of hMLH1 does not seem to account for all tumors with a MSI-positive phenotype in this population-based series of endometrial carcinomas. Our data indicate that the other mismatch repair genes hMSH2 and hMSH6 are also involved, especially in cases with intermediate MSI.     

2-DE protein expression in endometrial carcinoma

The objective of this study was to explore the protein expression pattern in normal endometrial mucosa (n = 5) and endometrial carcinoma (n = 15) of low (diploid) and high (aneuploid) malignancy potential by two-dimensional gel electrophoresis (2-DE). The specimens were evaluated for histopathologic subtype, stage and grade in relation to DNA ploidy. A match-set consisting of five samples from normal endometrium, eight diploid and seven aneuploid tumours was created. All the diploid and three of the aneuploid tumours were of endometrioid subtype, while the remaining four were of uterine seropapillary type. There were 192 protein spots differentiating diploid tumours from normal endometrium and 238 protein spots were separating aneuploid tumours from normal endometrium (p < 0.01). A cluster analysis based on 52 significantly deviating protein spots within the groups showed clustering and separation of the normal endometrium, diploid and aneuploid tumours. In conclusion this study showed significant differences in protein expression between normal endometrium and endometrial carcinoma as well as between endometrial carcinoma of low and high malignancy potential. In future studies these results may provide useful in finding new sensitive prognostic markers for endometrial cancer.     

散发性子宫内膜癌组织中错配修复基因hMLH1表达与微卫星不稳定性的相关性分析

目的：分析散发性子宫内膜癌组织中微卫星不稳定性（microsatellites instability,MSI）与错配修复基因hMLH1表达之间是否存在相关性。方法：应用免疫组化SP法检测40例子宫内膜癌、22例子宫内膜不典型增生、23例正常子宫内膜组织中错配修复基因hMLH1的表达;应用PCR方法检测子宫内膜癌中5个微卫星位点（D2S123、D10S197、D13S175、D10S215和D10S541）的微卫星不稳定性。结果：错配修复基因hMLH1在正常子宫内膜、子宫内膜不典型增生、子宫内膜癌组织中的表达逐渐下降,差异有统计学意义,χ^2=33.34,P=0.00;错配修复基因hMLH1蛋白的表达与内膜癌组织分化程度有关,χ^=7.98,P=0.02;hMLH1蛋白在内膜癌G1和G2期表达均为阳性;子宫内膜癌中微卫星不稳定性的发生与错配修复基因hMLH1的失表达密切相关,Pearson相关系数r=1,P=0.00。MSI在内膜癌组织中发生率显著高于正常子宫内膜,χ^2=5.26,P=0.02。结论：MSI是子宫内膜癌发生过程中重要的分子水平改变。hMLH1蛋白的表达有望成为筛查子宫内膜不典型增生向内膜癌发展的高危人群的一种手段,同时也可能成为判断内膜癌患者预后的指标。     

Activation of the receptor protein tyrosine kinase EphB4 in endometrial hyperplasia and endometrial carcinoma.

BACKGROUND: Members of the Eph family of tyrosine kinases have been implicated in embryonic pattern formation and vascular development; however, little is known about their role in the adult organism. We have observed estrogen-dependent EphB4 expression in the normal breast suggesting its implication in the hormone-controlled homeostasis of this organ. Since the endometrium is a similarly hormone dependent organ and endometrial carcinoma is thought to result from estrogenic stimulation, we have investigated EphB4 expression in normal human endometrium and during its carcinogenesis. PATIENTS AND METHODS: EphB4 expression was analyzed immunohistochemically in 26 normal endometrium specimens, 15 hyperplasias and 102 endometrioid adenocarcinomas and correlated with clinical and prognostic tumor characteristics. RESULTS: In normal endometrial tissue no EphB4 protein was detected. Strikingly, we observed a drastic increase (P <0.0001) in the number of EphB4 protein-expressing glandular epithelial cells in the majority of hyperplasias and carcinomas. Moreover, we found a statistically highly significant positive correlation between EphB4 expression and post-menopausal stage of the patient (P = 0.007). CONCLUSIONS: These findings indicate that in the endometrium, EphB4 is an early indicator of malignant development and, thus, EphB4 may represent a potent tool for diagnosis and therapeutic intervention.     

hMLH1 and hMSH2 expression in human hepatocellular carcinoma

The role of microsatellite instability (MSI) in the pathogenesis of hepatocellular carcinoma (HCC) is incompletely defined. Although high-frequency MSI (MSI-H) is infrequently seen in HCC, some studies have suggested a role for MSI in HCC development. While MSI has been clearly defined for a subset of tumors, in particular colorectal, gastric and endometrial cancers, generally accepted criteria have not been developed for other tumors. Colorectal cancers (CRC) are classified as MSI-H if >30-40% of >5 microsatellite loci analyzed show instability. The MSI-H phenotype is associated with defective DNA mismatch repair (MMR) and is observed in the majority of tumors from patients with hereditary non-polyposis colon cancer (HNPCC) and also in 15% of sporadic CRCs. Inactivating mutations of the hMLH1 or hMSH2 genes lead to defects in MMR in HNPCC. In sporadic CRCs, MMR is usually due to hypermethylation of the hMLH-1 promoter. The role of defective MMR in hepatocellular carcinogenesis is controversial. Immunohistochemistry for hMLH1 and hMSH2 reliably indicates hMLH1 or hMSH2 loss in MSI-H CRC tumors. To investigate the role of defective MMR in HCC carcinogenesis, we performed immunohistochemistry for hMLH1 and hMSH2 on 36 HCCs. BAT26, a microsatellite marker that reliably predicts MSI-H was also examined. All 36 of the tumors stained positively for both hMLH1 and hMSH2, strongly suggesting an absence of either inactivating mutations of hMLH1 and hMSH2 or promoter hypermethylation of hMLH1. None of the tumors showed MSI at the BAT26 locus. These findings suggest that defective MMR does not contribute significantly to hepatocellular carcinogenesis.     

散发性结直肠癌中p53蛋白表达与微卫星不稳定性的关系

目的 探讨散发性结直肠癌中微卫星不稳定性（ＭＩＮ）发生与Ｐ５３蛋白表达的相关性及其意义。方法：采用微卫星ＤＮＡ－ＰＣＲ－银染色法检测６７例散发性结直肠癌４条染色体上６个微卫星位点的ＭＩＮ,应用ＳＡＢＣ方法检测Ｐ５３蛋白表达。结果 以２个或２个以上位点有ＭＩＮ定义为复制误差阳性（ＰＣＲ＋）,ＲＥＰ＋率４１．８％（２８．．６７）。５０例Ｐ５３蛋白表达阳性６０．６７,７４．６％）。ＲＥＰ＋组Ｐ５３蛋白阳     

Negative hMSH2 protein expression in pancreatic carcinoma may predict a better prognosis of patients

The prognostic values of immunohistochemical staining for hMSH2 and hMLH1 in patients with pancreatic carcinomas were investigated. Fifty-five patients with histologically proven pancreatic carcinomas were studied. Immunohistochemical staining for hMSH2 and hMLH1 was performed by avidin-biotin-peroxidase complex method with a catalyzed signal amplification system. Tumor cells that exhibited an absence of nuclear staining in the presence of non-neoplastic cells with nuclear staining were considered to have an abnormal pattern. Four tumors (7.3%) demonstrated abnormal hMS2-negative staining. One tumor (1.8%) had abnormal hMLH1-negative staining and this tumor showed negative hMSH2 staining. No particular clinicopathologic factor such as tumor location, histology, T-factor, N-factor and TNM-stage were observed in patients with negative hMSH2 staining tumors. Median survival time (MST) of patients with hMSH2-negative tumors was comparatively longer than that of patients with hMSH2-positive tumors, however, there was no significant difference between MSTs of the two groups (56 months vs. 14 months; p=0.16). We concluded that negative hMSH2 immunohistochemical staining in the pancreatic carcinomas may demonstrate certain patients, but not all, who had microsatellite instability-positive tumors. The patients with hMSH2-negative tumors may have better prognoses than those with hMSH2-positive tumors. To clarify these findings, further large number of pancreatic carcinomas must be investigated.     

结直肠癌中FHIT蛋白的异常表达及其临床意义

目的探讨结直肠癌中脆性组氨酸三联体(FH IT)基因蛋白表达状况与临床病理指标的关系。方法采用半定量免疫印迹(W estern b lot)检测30例结直肠癌及其癌旁正常组织(≥5 cm)FH IT蛋白表达状况。结果30例癌组织有56.7%的FH IT蛋白表达异常,FH IT蛋白表达量与患者的年龄、性别、肿瘤部位、肿瘤大小及组织学类型无关(P>0.05)而与肿瘤的组织分化程度、浸润深度、Dukes分期和淋巴结转移有关(P<0.05)。在浸润深度越深、分化程度越低、Dukes分期越晚和有淋巴结转移的癌组织中,FH IT蛋白低表达越明显。结论FH IT蛋白表达状况可能与结直肠癌组织学分级、浸润程度、Dukes分期及淋巴结转移相关,提示FH IT蛋白表达降低可能对结直肠癌发生、发展具有重要作用。     

错配修复基因hMLH1和hMSH2在散发性大肠癌中的表达及其意义

目的 探讨错配修复基因hMLH1和hMSH2在散发性大肠癌(SCC)组织中的表达及其临床意义.方法 采用免疫组化Max Vision二步法对63例SCC标本中的癌组织、距癌灶3 cm以外的癌旁组织、距癌灶10 cm以外的正常组织中hMLH1和hMSH2蛋白的表达进行检测.结果 hMLH1蛋白在63例正常大肠组织、癌旁组织和SCC组织中的阳性表达率分别为95.2%、85.7%和81.0%,hMLH1蛋白在SCC组织中的阳性表达率明显低于正常大肠组织(P＜0.05).hMSH2蛋白在63例正常大肠组织、癌旁组织和SCC组织中的阳性表达率分别为76.2%、66.7%和52.4%,hMSH2蛋白在SCC组织中的阳性表达率明显低于正常大肠组织(P＜0.01).hMLH1蛋白在＜60岁的SCC患者组织中的阳性表达率(100%)明显高于≥60岁的SCC患者组织(75.0%,P＜0.05),在有淋巴结转移的SCC组织中的阳性表达率(50.0%)明显低于无淋巴结转移的SCC组织(93.3%,P＜0.05).hMSH2蛋白在＜60岁的SCC患者组织中的阳性表达率(80.0%)明显高于≥60岁的SCC患者组织(43.8%,P＜0.05),在癌组织浸润至肠壁浆膜层SCC组织中的阳性表达率(61.5%)明显高于浸润至黏膜下层和肌层的SCC组织(37.5%,P＜0.05).SCC组织中hMLH1和hMSH2蛋白的表达呈正相关关系(r=0.254,P＜0.01).结论 hMLH1和hMSH2蛋白在SCC组织中的表达均有一定的缺失,并且与患者的年龄、淋巴结转移和癌组织浸润的范围有关.hMLH1和hMSH2基因可以作为临床预测和判断SCC发生和发展有用的实验室指标.     

Microsatellite instability and immunohistochemical analysis of MLH1 and MSH2 in normal endometrium, endometrial hyperplasia and endometrial cancer from a hereditary nonpolyposis colorectal cancer patient

Hereditary nonpolyposis colorectal cancer (HNPCC)-related endometrial cancer is associated with mutations in DNA mismatch repair genes. However, chronological changes of these genes in the endometrium have not been studied in women from HNPCC families. Tissue samples of normal endometrium, endometrial hyperplasia without atypia and endometrial cancer were collected at different times from a 41-year-old Japanese woman with a family history of HNPCC. Combined microsatellite instability (MSI) and immunohistochemical analysis of MLH1 and MSH2 predicted the presence of a mutation in MSH2 when she had endometrial hyperplasia without atypia 7 months before the diagnosis of endometrial cancer. Endometrial hyperplasia without atypia may indicate an early development of endometrial cancer in women from HNPCC families.     

p53和hMSH2蛋白在甲状腺乳头状癌组织表达的意义

目的:探讨抑癌基因p53和错配修复基因hMSH2 蛋白在甲状腺乳头状癌和结节性甲状腺肿(简称结甲)组织中表达的意义.方法:用免疫组织化学链霉菌抗生物素过氧化物酶(SP)法检测65例甲状腺乳头状癌及其21例癌旁结甲、14例单纯性结甲和7例甲状腺瘤组织中p53和 hMSH2 蛋白的表达.结果:p53蛋白在甲状腺乳头状癌组织中表达率35.38%明显高于癌旁结甲9.52%和单纯结甲7.14%(P＜0.05),但与甲状腺瘤42.86%无显著性差异; p53 蛋白在有淋巴结转移组表达率53.85%明显高于无淋巴结转移组30.77%(P＜0.05).hMSH2 蛋白在甲状腺乳头状癌组织表达率72.31%明显高于癌旁结甲52.38%(P＜0.05)、单纯结甲21.43%(P＜0.01)和甲状腺瘤28.57%(P<0.01);hMSH2 蛋白在有淋巴结转移组表达率76.92%与无淋巴结转移组71.15%无显著性差异(P＞0.05).p53 蛋白阳性组甲状腺乳头状癌hMSH2蛋白阳性表达率82.61%,阴性组66.67%,两种蛋白表达呈正相关(P＜0.05).结论:甲状腺癌细胞的异常增生可能导致碱基错配的增多继而导致p53突变的积累,是甲状腺乳头状癌发生的原因之一,hMSH2 蛋白高表达可能是甲状腺癌发生的一个早期标志.     

Microsatellite instability and hMLH1/hMSH2 expression in young endometrial carcinoma patients: associations with family history and histopathology

Endometrial cancer is the second most common malignancy in patients with hereditary nonpolyposis colorectal cancer (HNPCC). The age at diagnosis of HNPCC-associated endometrial cancer is approximately 15 years younger than for sporadic endometrial cancer. Our current study was undertaken to determine the frequency of microsatellite instability (MSI) and absence of hMLH1 or hMSH2 protein expression in young patients with endometrial carcinoma and to correlate these findings with histopathologic and clinical features. Endometrial carcinoma from 62 women (23-52 years, median age 46) were assessed for MSI. Twenty-one of the 62 (34%) tumors demonstrated MSI. Of the 21 tumors demonstrating MSI, 12 showed an absence of hMLH1 expression, 4 showed an absence of hMSH2 expression, and 5 demonstrated normal expression of both proteins. All 41 tumors without MSI demonstrated normal hMLH1 and hMSH2 expression. Two patients with MSI tumors fulfilled the Amsterdam criteria for HNPCC, while 2 had histories suggestive of HNPCC. None of the patients with tumors without MSI had a personal or family cancer history suggestive of HNPCC. The MSI phenotype was associated (p < 0.05) with high FIGO stage and grade, cribriform growth pattern, mucinous differentiation and necrosis. Our findings suggest that the frequency of HNPCC in young endometrial cancer patients is relatively low when compared with the frequency of HNPCC in young colorectal cancer patients. Defects of the MMR proteins hMSH2 or hMLH1 account for MSI in most but not all endometrial cancers from young patients.     

子宫内膜癌中PTEN蛋白、P-AKT蛋白的表达及意义

目的:检测子宫内膜癌组织中PTEN蛋白及P-AKT蛋白表达并探讨其意义.方法:采用免疫组织化学法检测66例子宫内膜癌、28例正常子宫内膜组织中PTEN蛋白及P-AKT的表达.结果:子宫内膜癌组织中PTEN蛋白为缺失表达,高于正常子宫内膜组织(P＜0.05),而且PTEN表达在G1级肿瘤高于G2、G3级(P＜0.05),PTEN蛋白缺失率与肿瘤组织类型有关(P＜0.05),与肌层浸润、淋巴结转移及病理分期无明显关系(P＞0.05).子宫内膜癌组织中P-AKT蛋白表达为高表达,P-AKT的表达与肿瘤组织学分级、病理学分期、有无淋巴结转移以及肿瘤浸润子宫肌层深度显著相关(P＜0.05),其表达与组织学类型无关(P＞0.05).P-AKT与PTEN的表达呈明显负相关.结论:PTEN表达缺失与临床病理参数无关,蛋白表达缺失常发生细胞分化较差的子宫内膜癌.PTEN蛋白表达缺失与P-AKT表达水平有关.     

The expression of syndecan-1 is related to the risk of endometrial hyperplasia progressing to endometrial carcinoma

Objective

Aberrant expression of the cell surface proteoglycan, syndecan-1, is found in many malignancies. The current study describes the immunohistochemical study of syndecan-1 expression in normal, hyperplastic, and malignant endometrial tissues for evaluation of application as a parameter of cancer progression in patients with endometrial hyperplasia.

Methods

Immunohistochemical staining of syndecan-1 was performed in 101 formalin fixed, paraffin embedded sections of normal, hyperplastic, and malignant endometrial tissues. We analyzed specimens from patients with normal endometrium (NE, N=10) as controls, and those of simple hyperplasia (SH, N=20), complex hyperplasia without atypia (CH, N=20), atypical hyperplasia (AH, N=20), and endometrial cancer (EC, N=31).

Results

The mean rank of expression scores based on the frequency of syndecan-1 staining were 31.6, 20.5, 52.9, 72.1, and 62.1 for NE, SH, CH, AH and EC, respectively (p<0.001). Syndecan-1 expression was significantly greater in CH (p<0.001) or AH (p<0.001) than in SH, and significantly greater in AH compared to CH (p=0.028). Syndecan-1 is more frequently expressed in CH (p=0.042), AH (p<0.001), or EC (p=0.002) than in NE. Syndecan-1 expression did not differ significantly between NE and SH (p=0.248).

Conclusion

Syndecan-1 expression appears to be useful as a predictive indicator in endometrial hyperplasia.     

子宫内膜癌中EGFR蛋白的表达及EGFR基因突变

目的:研究表皮生长因子受体(epidermal growth factor receptor,EGFR)在子宫内膜癌组织中的表达及其基因突变情况。方法:取江西省肿瘤医院及南昌大学第二附属医院2007年1月至2011年4月期间的子宫内膜石蜡包埋组织104例,其中子宫内膜癌组织56例、非典型增生子宫内膜组织18例、正常子宫内膜组织30例,利用免疫组化检测上述各组织中EGFR蛋白的表达,PCR法扩增子宫内膜癌或正常子宫内膜组织中EGFR基因外显子19、21序列并测序检测其突变情况。结果:子宫内膜癌组织中EGFR的阳性表达率高于正常子宫内膜[73.2%(41/56)vs 30.0%(9/30),P<0.01]及非典型增生子宫内膜[73.2%(41/56)vs 44.4%(8/18),P<0.05]。进一步分析表明:G3级子宫内膜癌中EGFR的阳性表达率明显高于G1级[81.8%(9/11)vs 66.7%(12/18),P<0.01]及G2级[81.8%(9/11)vs 74.1%(20/27),P<0.05],在肌层浸润>1/2组中的阳性表达率也明显高于肌层浸润≤1/2组[86.8%(33/38)vs 44.4%(8/18),P<0.01]。但EGFR的表达与子宫内膜癌的FIGO分期、有无淋巴结转移无明显相关性。发现1例子宫内膜癌组织中,EGFR基因外显子19有G2281A突变,而外显子21无突变。结论:EGFR的阳性表达率与子宫内膜癌的组织学分级及肌层浸润深度有关,子宫内膜癌组中存在EGFR外显子19的G2281A点突变。