Coexpression index of estrogen receptor alpha mRNA isoforms in simple, complex hyperplasia without atypia, complex atypical hyperplasia and adenocarcinoma

OBJECTIVE: Estrogen receptor isoforms are postulated to play an important role in modulating the estrogen response. To clarify the molecular mechanisms through which malignant changes are activated in endometrium, this study aims to examine the expression profiles of wild-type ER-alpha and their splice variants and to assess the number of coexisting mRNA isoforms of ER-alpha in normal endometrium as well as in endometrial hyperplasia and endometrial endometrioid adenocarcinoma. METHODS: Human endometrium and specimens including endometrial hyperplasia and endometrial cancer were obtained during surgery. Endometrial data were classified into four groups: simple hyperplasia (n=24), complex hyperplasia (n=15), atypical hyperplasia (n=11), endometrial endometrioid adenocarcinoma (n=19) (grade 1, grade 2 morphological degree) and proliferative endometrium (n=24) as a control group. Total cellular RNA was extracted from endometrial tissues using Total RNA Prep Plus. A real-time quantitative RT-PCR assay was developed to quantify the wild-type ER-alpha and ER-alpha mRNA isoforms copy numbers. We have evaluated the variation in ERs mRNA level between normal endometrium and endometrial hyperplasia and adenocarcinoma. We also evaluated the "sharing indicator". It is a factor of mRNA ER-alpha holding shares in whole mRNA it assume quotient of ER-alpha slicing variant to all variants of mRNA ER-alpha. RESULTS: It was found that the number of coexisting mRNA isoforms was significantly higher in adenocarcinoma endometrium than that evaluated for various degrees of hyperplasia endometrium and normal proliferative endometrium (p<0.05, the Kruskal-Wallis test). CONCLUSION: The risk for progression of endometrial hyperplasia to atypical hyperplasia and eventually endometrioid adenocarcinoma may be accompanied by an increase in the number of alternative splicing variants of mRNA ER-alpha.     

Expression of cathepsin L in normal endometrium and endometrial cancer

OBJECTIVE: To evaluate the expression of cathepsin L in normal endometrium and endometrial adenocarcinoma. STUDY DESIGN: Tissue from eight cases of G1 and eight of G2 endometrioid adenocarcinoma, and 15 normal endometrial specimens were examined by immunohistochemistry. RESULTS: In the normal endometrium, cathepsin L was expressed in a few cell layers of the apical part of the glandular epithelium throughout the menstrual cycle. In the carcinomas, there was an inverse correlation between the grade of tumor and the cathepsin L expression. CONCLUSION: Cathepsin L expression may cease during endometrial carcinogenesis and its expression may be less important in tumor progression than it is in tumors of other tissues.     

Correlation of survivin mRNA detection with histologic diagnosis in normal endometrium and endometrial carcinoma

BACKGROUND: The objective of this study was to determine if survivin mRNA expression is a marker of endometrioid adenocarcinoma and if survivin mRNA levels correlate with tumor grade and stage. METHODS: Twenty-six samples of endometrioid adenocarcinoma and 18 cases of benign endometrium were obtained at surgery. RNA was extracted from tissues and was used for quantitative real-time RT-PCR, targeting a 91-bp sequence of survivin mRNA and the levels were standardized to the levels of ribosomal RNA. Statistical analysis of the correlation between histologic diagnosis and the corrected survivin mRNA levels was performed by the Fisher Exact test and the Kruskal-Wallis test. RESULTS: Survivin mRNA was detected in all specimens. Survivin mRNA levels were increased in proliferative endometrium (P = 0.0509) and was increased in correlation with ascending grade in endometrioid adenocarcinoma (P = 0.01). CONCLUSION: Survivin mRNA is not a specific marker of endometrial cancer, but may reflect an important mechanism in tumor progression of the endometrial mucosa.     

微管相关蛋白LC3和组织蛋白酶D在子宫内膜样腺癌中的表达及意义

目的:探讨微管相关蛋白LC3、组织蛋白酶D在子宫内膜样腺癌组织中的表达及与临床病理参数之间的关系。方法:用免疫组织化学法检测12例正常增生期子宫内膜、12例子宫内膜增殖症以及51例子宫内膜样腺癌组织中LC3和组织蛋白酶D的表达并分析。结果:LC3在子宫内膜样腺癌组织的表达强度显著低于正常增生期内膜和子宫内膜增殖症(P＜0．001,P＜0．001) LC3在子宫内膜样腺癌中的表达强度与组织学分级以及手术病理分期呈明显的负相关(r=-0．390,P＜0．001;r=-0．312,P＜0．05)。组织蛋白酶D在子宫内膜样腺癌组织中的表达显著低于正常增生期内膜(P＜0．05)。结论:自噬活性相关的LC3、组织蛋白酶D在子宫内膜样腺癌中的表达下降,自噬活性的改变可能参与了子宫内膜样腺癌的发生和演进过程。     

子宫内膜增生过长和子宫内膜样腺癌中VEGF 和TSP-1表达与血管新生研究

目的探讨子宫内膜增生过长及子宫内膜样腺癌中血管内皮生长因子(VEGF)和血小板反应素1(TSP-1)表达与血管新生的关系以及对子宫内膜样腺癌的发生、发展中的作用.方法采用免疫组织化学方法分别检测子宫内膜正常(12例)、增生过长(13例)、不典型增生(18例)及子宫内膜样腺癌(50例)中微血管密度(MVD)、VEGF及TSP-1表达情况.结果子宫内膜不典型增生和内膜样腺癌中MVD明显大于内膜正常及增生过长者(P＜0.05),子宫内膜样腺癌IA期与不典型增生二者的MVD差异无显著性(P＞0.05),而与正常内膜和增生过长者差异有显著性(P＜0.05);VEGF表达与上述不同内膜病变中MVD呈正相关(r=0.843,P=0.000 1),而TSP-1表达仅在子宫内膜样腺癌中分别与MVD和VEGF呈负相关趋势(r=-0.233,P=0.1041;r=-0.235,P=0.100 3);在子宫内膜样腺癌中TSP-1间质高表达且具有异质性.结论子宫内膜不典型增生和子宫内膜样腺癌中VEGF对血管新生起正向调节作用;TSP-1在部分腺癌患者中表达增强,但其负向调节作用较弱,可能会使血管新生开关平衡失调,与子宫内膜样腺癌发生及发展有关.     

skp2和p27~(kip1)在子宫内膜样癌组织中的表达

目的:检测子宫内膜样癌组织中skp2和p27kip1mRNA及其蛋白的表达,探讨其在子宫内膜样癌发生、发展中的意义。方法:采用半定量逆转录聚合酶链反应法(RT-PCR)检测37例子宫内膜样癌组织中skp2mRNA和p27kip1mRNA的表达,并进行半定量分析;应用免疫组化(PV-6000)二步法检测skp2和p27kip1蛋白的表达,以24例正常子宫内膜作为对照。结果:skp2mRNA平均表达水平在子宫内膜样癌组织中为0.79±0.10,正常子宫内膜为0.24±0.02,两者比较差异有统计学意义(P<0.01)。p27kip1mR-NA在子宫内膜样癌组织和正常内膜组织中表达无明显差异。子宫内膜样癌组织中skp2蛋白阳性表达率为78.38%,高于正常子宫内膜组织的4.17%,两者比较差异有统计学意义(P<0.01)。p27kip1蛋白在内膜癌组织阳性表达率低于正常内膜组织(48.65%vs79.17%),差异有统计学意义(P<0.05)。skp2 mRNA及蛋白表达与病理分级、淋巴结转移及肌层浸润深度呈明显正相关(P<0.05);p27kip1蛋白表达则与病理分级、淋巴结转移及肌层浸润深度呈明显负相关(P<0.05),两者均与年龄及手术病理分期无关(P>0.05)。子宫内膜样癌组织中skp2蛋白表达与p27kip1蛋白表达呈负相关(P<0.05)。结论:子宫内膜样癌组织中skp2 mRNA及蛋白高表达,p27kip1蛋白低表达,参与了子宫内膜样癌的发生、发展过程,并有可能成为子宫内膜样癌的早期诊断和治疗的新靶点。     

Expression of matrix metalloproteinase-26 and tissue inhibitors of metalloproteinases TIMP-3 and -4 in benign endometrium and endometrial cancer

OBJECTIVE: Matrix metalloproteinases (MMPs) and their physiological inhibitors, the tissue inhibitors of MMPs (TIMPs), play a key role in tumor cell invasion, angiogenesis, and growth. The aim of this study was to determine the expression and cellular distribution of MMP-26, TIMP-3, and TIMP-4 in endometrial cancers and benign endometrium throughout the menstrual cycle and the correlation with tumor histological subtype, stage, and grade. METHODS: Immunohistochemical analysis using polyclonal antibodies generated against pro- and active MMP-26, and mono- and polyclonal antibodies specific to TIMP-3 and TIMP-4, respectively, was performed. RESULTS: MMP-26, TIMP-3, and TIMP-4 are expressed in endometrial carcinomas (N = 86) and benign endometrium (N = 50) from various stages of the menstrual cycle. Semi-quantitative analysis of staining intensity indicated that endometrial carcinomas expressed more MMP-26, TIMP-3, and TIMP-4 compared to benign endometrium from the postmenopausal period, but not from the secretory phase of the menstrual cycle. The highest staining intensity was associated with endometrial epithelial cells, followed by vascular endothelial cells, myometrial smooth muscle cells, and endometrial stromal cells. Increased staining intensity of MMP-26 and TIMP-3 correlated with grade III tumors and MMP-26 and TIMP-4 with the depth of myometrial invasion in tumors histologically characterized as endometrioid adenocarcinoma, clear-cell, and papillary serous carcinoma staged/graded based on FIGO criteria. CONCLUSION: MMP-26 and TIMP-4 are expressed in endometrium and endometrial carcinoma and their elevated expression and correlation with myometrial invasion suggests that MMP-26 and TIMP-4 may play a key role in endometrial tumor progression.     

Expression of IGF-1R, VEGF-C and D2-40 and their correlation with lymph node metastasis in endometrial adenocarcinoma

Insulin-like growth factor-1 receptor (IGF-1R) regulates carcinogenesis and tumor development and is expressed in normal endometrium. Vascular endothelial growth factor C (VEGF-C) promotes lymph node metastasis. We investigated IGF-1R, VEGF-C and D2-40 in endometrial adenocarcinoma and the association between IGF-1R and lymphatic metastasis, using an immunohistochemical S-P method with 40 cases of endometrial adenocarcinoma and 14 of normal endometrium. IGF-1R expression differed significantly between normal endometrium and adenocarcinoma; it was associated with histological grade but not surgical stage. IGF-1R overexpression was associated with metastasis, but expression was not. VEGF-C expression was greater in normal endometrium than in adenocarcinoma and was associated with metastasis but not with surgical stage or histological grade. IGF-IR and VEGF-C expression were correlated in endometrial adenocarcinoma, and lymphatic vessel density was closely related to both. Abnormal IGF-IR and VEGF-C expression may be important in lymph node metastasis of endometrial adenocarcinoma and might be used to evaluate the prognosis.     

VEGF在绝经后子宫内膜样腺癌中的表达及临床意义

目的:探讨血管内皮生长因子(VEGF)在绝经后子宫内膜样腺癌中的表达及临床意义。方法:选择我院30例绝经后子宫内膜样腺癌组织和30例绝经后正常子宫内膜组织,应用免疫组织化学、原位杂交组织化学分别从细胞蛋白水平和分子基因水平检测VEGF在绝经后子宫内膜样腺癌组织的表达。结果:VEGF在绝经后子宫内膜样腺癌组织的表达显著高于正常子宫内膜组织(P=0.000);VEGF与组织学分级相关:其表达按高、中、低分化顺序升高(P=0.007,P=0.001);但与病理分期无关(P=0.160)。结论:VEGF在绝经后子宫内膜样腺癌高表达,参与绝经后子宫内膜样腺癌的肿瘤血管形成,可能在该疾病的早期发生和发展中发挥重要作用。     

Expression of survivin, PTEN and p27 in normal, hyperplastic, and carcinomatous endometrium

We aimed to investigate if expressions of survivin and p27 proteins are involved in the development of endometrioid carcinoma, along with whether there are any correlations between these proteins and loss of wild-type PTEN that is found in up to 80% of endometrial carcinomas. We also studied their correlations with classical prognostic factors and survival in endometrial carcinoma. To our knowledge, this is the first time survivin expression is investigated in endometrial hyperplasia along with endometrioid adenocarcinoma. For immunohistochemical analysis, 29 endometrioid adenocarcinoma, 38 endometrial hyperplasia, and 10 proliferative endometrium tissue samples were selected in the pathology archives. Staining of cells was scored as +2 if >50%, +1 if <50%, and negative if none were stained positive. Survivin expression increased from proliferative to hyperplasia to carcinoma cases. PTEN and p27 expressions decreased in hyperplasia and carcinoma cases with respect to proliferative endometrium. All these differences were statistically significant (P < 0.05). PTEN positively correlated to p27 (P < 0.05); however, neither was correlated with survivin. None of these genes were correlated with classical prognostic factors such as grade and myometrial invasion in endometrioid adenocarcinoma. However, mean survival was statistically significantly higher in PTEN-positive cases (46.6 vs 16.4 months) (P < 0.05). Survivin overexpression might be one of the important mechanisms in the development of endometrioid adenocarcinoma along with lost or decreased activity of PTEN and p27. However, survivin seems to exert its role in ways different from those of PTEN or p27 in the development of endometrioid adenocarcinoma. These findings on the role of survivin in endometrioid adenocarcinoma should be confirmed and the pathways through which survivin acts in endometrioid adenocarcinoma studied further with a larger sample size.     

HIF1α、Glut1在子宫内膜癌中的表达及其临床意义

目的:探讨HIF1α、G lut1在子宫内膜癌中的表达及其临床意义,并分析两者的相关性。方法:用免疫组化S-P法检测15例正常子宫内膜、17例不典型增生子宫内膜、45例子宫内膜癌组织中HIF1α、G lut1的表达。结果:HIF1α、G lut1在子宫内膜癌和不典型增生子宫内膜中的阳性率明显高于正常子宫内膜,差异有统计学意义(P<0.05)。在子宫内膜癌组织中HIF1α蛋白阳性表达率与组织病理学分级、手术病理分期无关(P>0.05),但与肌层浸润深度和淋巴结转移有关,差异有统计学意义(P<0.05);G lut1随病理组织学分级的增加、手术病理分期的进展、肌层浸润深度的加深和淋巴结转移,其阳性率逐渐上升,差异有统计学意义(P<0.05);HIF1α蛋白、G lut1蛋白表达呈正相关(r=0.741,P=0.000)。结论:HIF1α、G lut1蛋白的异常表达与子宫内膜癌的发生、发展有一定的关系。     

Heparanase expression increases throughout the endometrial hyperplasia-cancer sequence.

OBJECTIVE: To assess the expression of heparanase in the different stages leading to endometrial cancer. METHODS: The 38 examined specimens included adenocarcinoma, hyperplasia, and normal endometrium specimens. Heparanase, estrogen, and progesterone receptor expressions were analyzed immunohistochemically and the intensity was scored. RESULTS: Secretory normal endometrium and simple hyperplasia specimens expressed the lowest mean values of expression (1.00 and 0.63, respectively); the complex hyperplasia specimens and G2 endometrioid adenocarcinoma showed the highest values of expression (2.33 and 2.71, respectively). A linear trend (P=0.005) of heparanase expression was observed when comparing the normal endometrium and simple hyperplasia group with the complex hyperplasia+G1 carcinoma group and the G2+G3 carcinoma group. Evaluation of atrophic and inactive endometrium compared with papillary serous carcinomas yielded no significant differences. We found no significant correlation between heparanase expression and estrogen receptor or progesterone receptor expression. CONCLUSION: Heparanase expression was tightly regulated in endometrial tumorigenesis.     

Apoptosis and the expression of Bcl-2 and Bax in patients with endometrioid, clear cell, and serous carcinomas of the uterine endometrium

OBJECTIVES: Clear cell and serous carcinoma of the uterus are rare types of endometrial carcinomas. This study was designed to investigate the differential occurrence of apoptosis, Bcl-2, and Bax in endometrioid, clear cell, and serous carcinomas. METHODS: In a total of 28 endometrial carcinomas as well as 4 samples of normal postmenopausal endometria, apoptotic changes were examined using molecular biochemical techniques. The expression of Bcl-2 and Bax proteins was also investigated by immunohistochemical staining with appropriate antibodies. RESULTS: Labeling of DNA in situ indicated that apoptotic cells were sporadically seen in postmenopausal endometrium (5.2 +/- 2.1, n = 4). In contrast, cells undergoing apoptosis apparently were detected in endometrioid carcinoma (29.3 +/- 3.7, n = 20), and their numbers increased intensely in clear cell (49.5 +/- 5.6, n = 5) and serous carcinomas (50.8 +/- 6.0, n = 3). Autoradiographic analysis revealed that high-molecular-weight DNA was predominant in postmenopausal endometrium. However, a DNA ladder was identified in 7 of 10 carcinomas. Although Bcl-2 was immunonegative or faintly immunopositive in all cases, many cases of endometrioid carcinoma (43.6 +/- 4.1%, n = 20) were immunopositive for Bax, unlike postmenopausal endometrium (17.6 +/- 6.7%, n = 4). Moreover, the number of cells expressing Bax increased in clear cell (60.4 +/- 6.5%, n = 5) and serous carcinomas (66.8 +/- 7.6%, n = 3) compared with that in endometrioid carcinoma. CONCLUSIONS: These results indicate that apoptosis occurs in a specific population of cells in different histologic components of endometrial carcinomas. The expression of Bax, but not of Bcl-2, might suggest histologic differentiation in endometrial carcinomas.     

肿瘤浸润性树突状细胞在子宫内膜癌中的表达及其临床意义

目的探讨不同病理分级、临床分期的子宫内膜癌中肿瘤浸润性树突状细胞(TIDC)临床表达及其临床意义。方法 2005年1月至2009年10月在暨南大学附属第一医院对45例子宫内膜癌组织中TIDC行S-100单克隆抗体免疫组织化学染色,通过体视学参数分析TIDC在子宫内膜癌中的浸润状况;采用流式细胞仪检测TIDC膜表面MHCⅡ-分子及CD54的表达。结果子宫内膜癌中TIDC数量显著少于正常子宫内膜组织中树状突细胞(DC)数量(数密度参数比为:9.81±1.03 vs 19.37±1.14,P<0.05);不同病理分级子宫内膜癌中TIDC数量有统计学意义(即高分化、中分化、低分化子宫内膜腺癌TIDC数密度参数比:11.67±1.98 vs 7.08±1.17 vs 3.36±0.28,P<0.05);不同临床分期子宫内膜癌中TIDC数量有统计学意义(Ⅰ期、Ⅱ期、Ⅲ~Ⅳ期子宫内膜癌中TIDC数密度参数比:12.85±2.01 vs 6.89±1.07 vs 3.85±0.56,P<0.05);子宫内膜癌组织TIDC膜表面MHCⅡ-分子、CD54的表达显著低于正常子宫内膜组织[(5.92±1.14)%vs(11.09±1...     

Expression of fos and jun proto-oncogenes in benign versus malignant human uterine tissue

OBJECTIVE: The objective of this study was to evaluate expression of fos and jun proto-oncogenes in benign human uterine tissue compared with malignant uterine tissue. METHODS: Forty-two endometrial tissue specimens were obtained at the time of hysterectomy. Tissue samples from different phases of the menstrual cycle and from postmenopausal patients were stained using immunohistochemical methods to detect Fos and Jun proteins, estrogen and progesterone receptor status, and Ki67 (detects a nuclear antigen associated with proliferating cells). Tissue was examined microscopically for nuclear staining in endometrial epithelium and stroma. The endometrium was based on the patient's last menstrual period, pathologic dating, and proliferative versus nonproliferative status as determined by Ki67. Benign and malignant specimens were subjected to Northern blot analysis to evaluate levels of expression of c-fos, c-jun, and jun-B mRNA. The pattern of c-fos mRNA expression in malignant samples was further evaluated using in situ hybridization. RESULTS: In proliferative, secretory, postmenopausal, and progesterone-influenced, uterine specimens immunohistochemically stained and examined, the endometrial and stromal nuclei stained for both Fos and Jun in varying intensities. However, no pattern was found in the variation of intensity according to the phase of the endometrium. Similarly, in malignant and benign endometrial tissue examined by Northern blot and in situ hybridization analyses, expression of proto-oncogene mRNAs was readily detectable, but no statistical correlation between type of tissue examined, grade of adenocarcinoma, and stage of endometrial cancer was found in this study. CONCLUSIONS: In rodent models, control of uterine cell proliferation is related to change in expression of fos and jun proto-oncogenes. Our results indicate that hormonal control is likely to be different in human endometrium and probably involves genes other than the proto-oncogenes under study. Expression of Fos and Jun do not correlate with endometrial cancer stage and grade.     

Expression of aquaporin-1 in normal, hyperplasic, and carcinomatous endometria

OBJECTIVE: To explore the relationship between aquaporin-1 (AQP1) and endometrial adenocarcinoma. METHOD: Intratumoral microvessel density (IMD) was assessed as well as AQP1 and vascular endothelial growth factor expression in samples from 117 women, 75 with endometrioid adenocarcinoma, 17 with endometrial hyperplasia, and 25 with normal proliferative endometria. RESULTS: AQP1 was located in the epithelial cells of microvessels and small vessels in all samples. The AQP1/IMD ratio was highest in samples from the first, less in samples from the second, and least in samples from the third group. In samples from endometrioid adenocarcinoma, the AQP1/IMD ratio was significantly correlated with histologic grade, surgical stage, myometrial invasion, and extrauterine metastasis. There was a positive correlation between AQP1 expression and IMD and between AQP1/IMD ratio and VEGF expression. CONCLUSION: AQP1 may be involved in the tumorigenesis and progression of endometrioid adenocarcinoma by promoting angiogenesis, and AQP1 level may be both a tumor indicator and a new therapeutic target.     

The clinical significance of tumor-associated antigen RCAS1 expression in the normal, hyperplastic, and malignant uterine endometrium

OBJECTIVE: A tumor-associated antigen, RCAS1, is recognized by 22-1-1 monoclonal antibody. It was found in carcinomas derived from the uterus and ovary and was especially strongly expressed in invasive cancers. A previous investigation showed the RCAS1 expression to be correlated with a poor prognosis in uterine cervical adenocarcinoma. In this study, we examined whether the expression of RCAS1 is associated with the progression of the uterine endometrial neoplasms. METHODS: The expression of RCAS1 was evaluated by an immunohistochemical analysis. The tissue specimens used in this study included 46 cases of normal uterine endometrium, 40 cases of hyperplasia, and 121 cases of adenocarcinoma. The relationship between RCAS1 expression and several clinicopathological variables (clinical stage, histology, grade, myometrial invasion, lymph-vascular space invasion, and lymph node metastasis) was also assessed in endometrial adenocarcinoma. RESULTS: RCAS1 was positive in 26% of the normal uterine endometrium specimens (12 of 46 total cases), in 32% of the hyperplasia specimens (13 of 40 total cases), and in 68% of the adenocarcinoma specimens (83 of 121 total cases). As a result, the expression of RCAS1 was statistically higher in adenocarcinoma than in the normal and hyperplastic endometrium (P < 0.0001). RCAS1 was statistically detected more frequently in grade 3 than in grade 1 or 2 (P < 0.05); however, there was no correlation between the antigen expression and the clinical stage, myometrial invasion, lymph-vascular space invasion, or lymph node metastasis. CONCLUSION: RCAS1 expression might thus be associated with the malignant transformation and poor differentiation observed in uterine endometrial adenocarcinoma.     

Grade 2 endometrioid adenocarcinoma arising from adenomyosis of the uterus: report of a case

Adenomyosis is defined by the presence of endometrial tissue (glands and stroma) within the myometrium and malignant transformation of adenomyosis in premenopausal women with normal endometrium is extremely rare. Adenocarcinomas arising within adenomyosis need to be distinguished from endometrial carcinomas which arise from the eutopic endometrium, then extend into preexisting adenomyosis of the uterine wall. We report a case of grade 2 endometrioid adenocarcinoma arising from an adenomyotic focus in the uterus.