钙调蛋白结合蛋白在子宫平滑肌肿瘤诊断中的意义

目的　探讨高度敏感和特异的诊断子宫平滑肌肿瘤的指标。方法　选取子宫内膜间质肉瘤 2 6例 ,子宫普通型和富于细胞性平滑肌瘤各 2 5例 ,子宫平滑肌肉瘤 17例 ,正常子宫肌层和正常子宫内膜各 2 5例 ,采用免疫组化 S-P法检测各种组织中结蛋白、平滑肌肌动蛋白、雌激素受体以及重型钙调蛋白结合蛋白 (h-caldesmon)的表达。结果　正常的子宫肌层、子宫普通型平滑肌瘤、富于细胞性平滑肌瘤和子宫平滑肌肉瘤中重型钙调蛋白结合蛋白的表达分别为 10 0 .0 %、10 0 .0 %、96.0 %和 64.7% ,但正常子宫内膜和子宫内膜间质肉瘤细胞中均未见其表达。重型钙调蛋白结合蛋白诊断富于细胞性平滑肌瘤不仅敏感度高于结蛋白 ,而且特异度达 10 0 .0 %。结论　重型钙调蛋白结合蛋白可辅助正确鉴别子宫富于细胞性平滑肌瘤和子宫内膜间质肉瘤     

Elevated levels and distinct patterns of expression of A-type cyclins and their associated cyclin-dependent kinases in male germ cell tumors

Aberrant expression of several key regulators controlling the G1/S phase of the cell cycle has been implicated in human male germ cell tumorigenesis. Given the critical role of cyclin A2 at both the G1/S and G2/M transitions and the essential role for cyclin A1 in male germ cell development, our present study focused on the involvement of the A-type cyclins in the transformation and progression of male germ cell tumors (GCTs). The expression of the A-type cyclins and their catalytic partners Cdk1 and Cdk2 was examined in all types and stages of human male GCTs, including carcinoma in situ(CIS), seminoma and non-seminoma GCTs, along with normal testis samples. Elevated levels of cyclin A2, Cdk1 and Cdk2 were detected in the majority of GCTs and were correlated with the invasiveness of the tumors (p < 0.05). Cyclin A1 expression was virtually undetectable in CIS and seminoma, but was aberrantly expressed in all non-seminomatous GCTs. Cyclin A2 expression was strongly correlated with that of its catalytic partners Cdk1 and Cdk2 in all types of testicular tumors examined (p < 0.05), whereas a strong correlation between cyclin A1 and Cdk1 or Cdk2 was only seen in non-seminomatous GCTs (p < 0.05). Histone kinase activities of cyclin A1/Cdks and cyclin A2/Cdks were found to be elevated in tumors. Our data suggest that aberrant expression of A-type cyclins and their Cdks is a significant factor in male germ cell tumorigenesis. The abundant ectopic expression of cyclin A1 in non-seminomatous GCTs and its absence in CIS and seminomas is likely linked to the tumor transformation and progression and may be relevant to clinical prognosis.     

Overexpression of G1-S cyclins and cyclin-dependent kinases during multistage human pancreatic duct cell carcinogenesis.

PURPOSE: Molecular analysis of pancreatic intraepithelial neoplasia lesions and ductal adenocarcinoma suggested a multistage paradigm for pancreatic duct cell carcinogenesis. This study investigated the molecular basis for the neoplastic duct cells in this pancreatic intraepithelial neoplasia-carcinoma sequence to acquire progressive enhancement of their proliferative potential. EXPERIMENTAL DESIGN: Using tissue microarray blocks containing 15 to 40 pancreatic intraepithelial neoplasia lesions and ductal adenocarcinoma of pancreas, we studied by immunohistochemistry the expression profiles of cyclins and cyclin dependent kinases (CDKs) that regulate the G1-S cell cycle checkpoints. The role of cyclins D3 and D1 in three pancreatic cancer cell lines was investigated using specific short interfering RNA technique. RESULTS: Cyclin D3 overexpression was noted the earliest in pancreatic intraepithelial neoplasia-1A and was prevalent in 90% to 100% of high-grade pancreatic intraepithelial neoplasias and ductal cancer. Cyclin A overexpression was also noted early and reached 50% to 100% of high-grade pancreatic intraepithelial neoplasias and cancer, but the percentage of abnormal duct cells showing overexpression of cyclin A was significantly lower than cyclin D3. Cyclin E overexpression occurred in 20% to 25% of high-grade pancreatic intraepithelial neoplasias and in 75% of ductal carcinoma. Cyclin D1 demonstrated the lowest frequency of overexpression that occurred late. CDK2 and CDK4 overexpression was also noted in early pancreatic intraepithelial neoplasias and progressively increased to reach 60% to 75% in carcinoma. The down-regulation of cyclin D3 mRNA and protein levels using specific short interfering RNA resulted in growth inhibition of pancreatic cancer cell lines. CONCLUSION: The results provide additional insight into the mechanism of G1-S cell cycle checkpoints deregulation during stepwise pancreatic duct cell carcinogenesis, and suggest a p16-independent role for cyclin D3 in deregulating the G1 cell cycle checkpoints during early stages of pancreatic duct cell carcinogenesis.     

Clinical implications of cyclins,cyclin-dependent kinases,RB and E2F1 in squamous-cell lung carcinoma

In the search for new risk factors at the molecular and cellular levels, clinical data [lymph-node involvement (LN) and stage] were used and 104 squamous-cell lung carcinomas were analyzed by immuno-histochemistry for expression of cyclin D1, cyclin A, cdk2, cdk4, RB, and E2F1. The results of the univariate analysis of all 8 factors showed that cyclin A and cdk2 gave the best prognostic information, while no prognostic value could be found associated with cyclin D1, cdk4, RB and E2F1. The subsequent multivariate analysis of all possible combinations of the important factors showed that the pairs LN/cyclin A, LN/cdk2 and cyclin A/cdk2, and the triplet LN/cyclin A/cdk2 yielded the best prognostic information. It was essentially better than the information given by a single factor. Int. J. Cancer (Pred. Oncol.) 79:294–299, 1998.© 1998 Wiley-Liss, Inc.     

Expression and CpG methylation of the insulin-like growth factor II gene in human smooth muscle tumors.

Previously we have shown that expression of the insulin-like growth factor II (IGF-II) gene in 36 normal smooth muscle tissues (myometria) and 26 benign smooth muscle tumors (leiomyomas) was detectable by Northern blot analysis but that the RNA levels were low. In 9 of 20 malignant smooth muscle tumors (leiomyosarcomas) IGF-II gene expression was also low or absent, while in 11 of 20 the IGF-II gene was abundantly expressed. In 32 of these tissues we have now studied the DNA methylation state of the IGF-II gene. For the analysis of overall methylation of the gene the restriction endonucleases HpaII and MspI were used. In normal smooth muscle and in leiomyomas the IGF-II gene appeared to be methylated. In leiomyosarcomas with low IGF-II gene expression the DNA was partly demethylated. In leiomyosarcomas with abundant IGF-II gene expression overall methylation of the DNA tended to be low. In addition, we have studied the methylation state of one particular CpG site in the IGF-II gene with the restriction endonuclease AvaII. The results of the latter analysis confirm the analysis with HpaII and MspI. In conclusion, in malignant smooth muscle tumors the data indicate an inverse correlation between CpG methylation and expression of the IGF-II gene.     

Mesenchymal tumors of the uterus. VI. Epithelioid smooth muscle tumors including leiomyoblastoma and clear-cell leiomyoma: a clinical and pathologic analysis of 26 cases.

Twenty-six cases of atypical smooth muscle tumors of the uterus, including leiomyoblastoma, epithelioid leiomyoma, clear-cell leiomyoma, and plexiform tumorlet, are presented. The characteristic microscopic feature serving as the basis for inclusion in this study is the rounded to polygonal shape of the majority of cells instead of the elongated blunt-ended shape of smooth muscle cells seen in the typical leiomyoma. Mixtures of epithelioid, clear-cell, and plexiform patterns occurred with sufficient frequency to indicate that such patterns are variants of a single entity. A transition to typical smooth muscle cells was found in most instances, confirming the smooth muscle origin of these tumors. Features related to a favorable prognosis include the presence of clear cells, an expansile tumor margin, extensive hyalinization, and absence of extensive necrosis as seen microscopically. Until additional evidence has been accumulated it is proposed that neoplasms having five or more mitotic figures per ten high-power fields (HPF) be tentatively termed epithelioid leiomyosarcoma or leiomyosarcoma with epithelioid features and those with less than five mitotic figures per ten HPF, epithelioid leiomyoma.     

Increased expression of G1 cyclins and cyclin-dependent kinases during tumor progression of chemically induced mouse skin neoplasms

Cyclins and cyclin-dependent kinases (Cdks) are central to regulation of the cell cycle. Their abnormal expression may cause loss of cell-cycle control and result in autonomous cell growth, a critical feature of neoplasias. In this study, using immunoblotting, we analyzed the protein levels of several G₁/S cyclins (cycling D1, D2, D3, A, and E) and their respective Cdks (Cdk 2, 4, and 6) in 17 mouse squamous cell carcinomas (SCCs) and 18 mouse skin tumor cell lines. Overexpression of these cell cycle-related genes was frequent in tumors and cell lines. Of special interest was the fact that a group of cell lines that became more aggressive after animal passaging expressed more cyclins D2 and D3 than their respective parental lines did. In addition, SCCs had higher cyclin D3 expression levels than papillomas, and metastases had higher levels than the respective primary tumors, indicating that overexpression of cyclin D3 may be associated with increased aggressiveness of mouse SCC. Interestingly, overexpression of cyclin E was seen in most SCCs induced by a complete carcinogenesis protocol with benzo[a]pyrene (B(a)P) and only in a few SCCs induced by a two-stage carcinogenesis protocol using 7,12-dimethylbenz[a]anthracene as initiator. In contrast, more of the latter tumors overexpressed cyclin D1 and D2 than those induced by B(a)P. Thus, it is possible that different components of the cell-cycle machinery are involved in proliferative dysfunctions that take place during tumor development with different carcinogenesis protocols. Taken together, these results indicate that overexpression of G₁ cyclins and their related Cdks is a significant molecular abnormality that could be involved in the process of tumor progression. Mol. Carcinog. 18:142–152, 1997. © 1997 Wiley-Liss, Inc.     

子宫平滑肌肿瘤组织bcl-2蛋白表达的研究

目的 :了解bcl 2蛋白在子宫平滑肌肿瘤 (USMTs)的表达和意义。方法 :在 2 0例良性子宫平滑肌瘤 (UL)、18例子宫平滑肌肉瘤 (LMS)及 70例交界性子宫平滑肌瘤(BLM )中 ,应用免疫组织化学法检测bcl 2蛋白的表达。结果 :UL组bcl 2蛋白表达阳性率明显高于LMS组 ,P <0 0 5 (18例 /10例 ) ;BLM组bcl 2蛋白表达阳性率显著高于LMS组 ,P <0 0 1(62例 /10例 )。结论 :bcl 2蛋白过度表达对细胞凋亡旁路的抑制可能在USMT的发生、发展起到重要的作用     

DNA计量图像分析诊断子宫平滑肌肿瘤的特异性及标准化检测

应用电子计算机辅助ＤＮＡ影像计量分析仪（ＣＭ１ＤＮＡＣｙｔｏｍｅｔｅｔ．ＨＵＮＤ，Ｗｅｔｚｌａｒ，Ｇｅｒｍａｎｙ）测量３０例人正常子宫平滑肌、内膜上皮、纤维及淋巴细胞核ＤＮＡＩＯＤ值，以确定子宫组织特异性正常二倍体参照细胞及其校正因子。测量８０例子宫平滑肌瘤细胞核ＤＮＡ含量，发现各ＤＮＡ计量结果与正常平滑肌近似，无１例出现干系ＤＮＡ非整倍体和＞９ｃＥＥ，有４例出现＞５ｃＥＥ。测量３２例生长活跃的子宫平滑肌瘤，１８例检出干系ＤＮＡ非整倍体，１例检出＞９ｃＥＥ，１１例检出＞５ｃＥＥ，测量２７例子宫平滑肌肉瘤，２２例出现干系ＤＮＡ非整倍体，５例出现＞９ｃＥＥ，１８例出现＞５ｃＥＥ。本文结果表明，用于系ＤＮＡ非整倍体和＞９ｃＥＥ作为指标诊断子宫平滑肌瘤的特异性均为１００％，诊断子宫平滑肌肉瘤的敏感性分别为８１％和１９％，对生长活跃的子宫平滑肌瘤恶性性质的预测具有重要意义。＞５ｃＥＥ这一指标阈值不适合用于对子宫平滑肌肿瘤良、恶性质的判断。     

p53和c-myc蛋白在子宫平滑肌肿瘤的表达及意义

目的：了解p53和c-myc蛋白在子宫平滑肌肿瘤(USMTs)的表达及意义。方法：收集本院20例良性子宫平滑肌瘤(UL)、18例子宫平滑肌肉瘤(LMS)及70例交界性子宫平滑肌瘤(BLM)，其中40例富于细胞型子宫平滑肌瘤(CL)、13例奇异型子宫平滑肌瘤(BL)、4例核分裂活跃型子宫平滑肌瘤(ML)、10例不典型子宫平滑肌瘤(AL)及3例恶性潜能未定型子宫平滑肌瘤(STUMP)应用免疫组化法检测p53和c-myc蛋白的表达。结果：肉瘤组p53蛋白表达阳性率明显高于肌瘤组(P＜0．01)和交界瘤组(P＜0．01)；肉瘤组c-myc蛋白表达阳性率显著高于肌瘤组(P＜0．01)和交界瘤组(P＜0．01)，c-myc蛋白表达形式在良、恶性USMT有显著不同(P＜0．01)。结论：肉瘤的发病机制涉及p53基因功能的丧失，p53蛋白表达增加提示USMT细胞恶性生物学行为的潜能增加。c-myc的过量表达与USMT的发生、发展密切有关，在良、恶性USMT表达形式有显著不同。     

Cell cyclins and cyclin-dependent kinase activities in mouse mammary tumor development

Breast cancer in humans, as in mice and rats, is thought tobe the result of sequential changes in the epithelial cellsof the mammalian glands. This study examines the altered expressionor activation of cell cycle related proteins in an in situ systemcomposed of hyperplasia, preneoplasia and neoplasia of mousemammary glands. The results showed a high level of cdc2/cdk2kinase activities in tumors compared to hyperplasias which wasindependent of cdc2/ cdk2 protein levels. Some of the cdk-associatedproteins which are thought to regulate cdk kinase activity wereexamined in these tissues. Cyclin A was overexpressed in allhyperplasias irrespective of their tumorigenic potentials. However,a number of alterations in cyclin E protein were associatedwith cdk2 and its associated kinase activity during mammarytumorigenesis. First, the level of normal cyclin E (p50) expressionwas positively correlated with the tumorigenic potentials ofdifferent hyperplasia lines. Second, several cyclin E isoforms(p48, p43, p35, p34, p32) were detected only in tumor tissues.Third, a 2.3- and 8.3-fold increase in cyclin E-associated cdk2kinase activity was present in highly tumorigenic hyperplasiasand neoplasias respectively compared to the low tumorigenichyperplasias. Polymorphic cell nuclear antigen (PCNA) proteinbound to cdk2 was a better indicator for cell proliferationand cdk2 kinase activity than the PCNA labeling index. Theseresults suggest a sequential pattern of multiple derangementsin factors regulating cdk2 protein function during mammary tumorigenesis.High levels of cdk2 kinase activity are observed only in tumorsand appear to be closely related to alterations in cyclin Eprotein expression.     

C-kit expression in uterine leiomyosarcomas: an immunocytochemical study of 29 cases of malignant smooth muscle tumors of the uterus

Uterine malignant stromal tumors are rare neoplasms characterized by fatal prognosis. At the moment no effective systemic treatment is available for metastases or recurrent disease. The drugs employed in advanced neoplasms are iposfamide, doxorubicin or epidoxorubicin, but the clinical response to chemotherapy is poor. Recent studies have shown that cells in gastrointestinal stromal tumors express a growth factor receptor with tyrosine kinase activity termed c-kit. Lately reports of efficacy of a specific anticancer drug with imatinib (ST1571) based on specific molecular abnormalities of proto-oncogene c-kit present in gastrointestinal stromal tumors induced us to identify the c-kit phenotype also in uterine leiomyosarcomas. These data may be useful for treating metastatic uterine leiomyosarcomas with increased c-kit kinase activity.     

Flow cytometric DNA analysis of gastric smooth muscle tumors.

BACKGROUND. To better understand the malignant grade of gastric smooth muscle tumors, the DNA content of these tumors was studied. METHODS. In 43 patients with gastric smooth muscle tumors, the cellular DNA content was determined by flow cytometry and compared with the histologic classification and the prognosis. RESULTS. Flow cytometry indicated that all 21 leiomyomas and 9 of the 10 low-grade leiomyosarcomas were diploid; 10 of the 12 high-grade leiomyosarcomas were aneuploid. All patients with leiomyomas and 8 of the 11 patients with diploid leiomyosarcomas had neither local recurrence nor metastasis. By contrast, 8 of the 10 patients with aneuploid leiomyosarcomas died of their disease (mean survival, 41 months; range, 18-78 months). CONCLUSIONS. These results indicate that the DNA ploidy pattern shown by flow cytometry is related closely to the histologic classification and prognosis of gastric smooth muscle tumors.     

Colorectal mesenchymal tumors - from smooth muscle tumors to stromal tumors

Colorectal mesenchymal tumors are rare. Therefore, distinguishing between gastrointestinal stromal (GIST) and smooth muscle tumors is important. This study aimed to delineate the immunophenotype and prognostic factors of 75 colorectal mesenchymal tumors. Fifty-three GIST and 22 smooth muscle tumor specimens were included from 1986 to 2007. Forty of 53 GIST were initially diagnosed as smooth muscle tumors and re-diagnosed as CD117 (+) GIST. Immunohistochemical studies were performed with antibodies of CD117, CD34, smooth muscle actin (SMA), desmin, S-100, Ki-67 and PCNA for clinicopathologic and prognostic correlation. In comparison, colorectal GIST exhibited a larger tumor size (P<0.001), higher mitotic count (P<0.001), higher cellularity (P<0.001), less spindle cell type (P=0.004), higher nuclear pleomorphism (P=0.004), and a higher NIH risk (P<0.001) than that of smooth muscle tumors. Positive immunoreactivities of GIST to a panel of antibodies were 88.6% to CD34, 28.3% to SMA, 1.8% to S-100 and 15.1% to desmin. For 75 mesenchymal tumors, survival analyses revealed that older patients (P=0.006), with a large tumor size (P<0.001), high mitotic count (P<0.001), increased NIH risk (P<0.001), non-spindle cell type (P<0.001), high cellularity (P=0.015), high cell pleomorphism (P<0.001), positive Ki-67 (P<0.001), high PCNA (P<0.001) and GIST (P=0.001) had a shorter disease-free survival than that of comparative groups. When the analyses concentrated on 53 GIST, the cell type and cellularity were no longer viable prognostic factors. The tumor mitotic count was the only independent prognostic factor for either mesenchymal tumors or GIST. In conclusion, GIST exhibited heterogeneous characteristics and was significantly larger, more mitotic and a poorer prognostic factor than smooth muscle tumor. The mitotic count is still the most valuable prognostic factor for colorectal mesenchymal tumors after KIT.     

食管间质瘤与平滑肌肿瘤的临床鉴别及治疗

背景与目的：食管间质瘤与平滑肌肿瘤虽然具有不同的病理学特征,但是临床上不易鉴别。本研究比较食管间质瘤和平滑肌肿瘤的临床特征,并探讨其治疗原则。方法：用免疫组织化学法检测原诊断为食管平滑肌（肉）瘤20例和间质瘤1例肿瘤组织中CD117和CD34等一组抗体表达。结合病理学形态表现和临床表现进行分析,并总结各自的治疗原则及疗效。结果：16例原诊断为食管平滑肌瘤有5例CD117（＋）,按照间质瘤评定标准,均为非高度侵袭危险性食管间质瘤,余11例CD117（-）;1例原间质瘤CD117（＋）和CD34（＋）,为高度侵袭危险性食管间质瘤;4例食管平滑肌肉瘤CD117（-）和CD34（-）。食管非高度侵袭危险性间质瘤和平滑肌瘤的病理学形态表现、临床表现、治疗方法和预后没有明显差异;食管高度侵袭危险性间质瘤和平滑肌肉瘤的病理学形态表现、临床表现、治疗方法和预后也没有明显差异。结论：病理学形态表现和临床表现无法区分食管间质瘤和平滑肌肿瘤,CD117等抗体的免疫组织化学法检测是区分食管间质瘤和平滑肌肿瘤必不可少的手段。食管非高度侵袭危险性间质瘤和平滑肌瘤局部切除术或部分食管切除术可以达到根治,预后良好;高度侵袭危险性食管间质瘤和平滑肌肉瘤需行食管部分切除术。     

Regulators of G1 cyclin-dependent kinases and cancers

The mammalian cell cycle can be divided into four phases: G1 (gap phase 1), S (DNA synthesis), G2 (gap phase 2), and M (mitosis). Progression through each phase of the cell cycle is delicately controled by the activity of different cyclin-dependent kinases (CDKs) and their regulatory subunits known as cyclins. CDK2, CDK4, CDK6 and their associated cyclins control the G1 to S phase transition. The association of CDK4 or CDK6 with D-type cyclins is critical for G1 phase progression, whereas the CDK2-cyclin E complex is important for initiation of the S phase. Cancer can originate from dysregulation of these regulators. A variety of intrinsic and extrinsic signals were recently identified to regulate these G1 or G1/S CDKs and cyclins. Here we discuss the regulators of these protein kinases at different mechanistic level with a hope that these insights can be applied to develop therapeutic strategies for cancer treatment.