Evaluation of topoisomerase II, ki-67, and P53 expression in non-muscle-invasive urothelial carcinoma and their clinical significance

Background: Transurethral resection of tumor is the main treatment of non-muscle-invasive urothelial carcinoma, but it is associated with high rate of recurrence and/or progression and this arouses the need for adjuvant therapy. Topoisomerase II (Top II), KI-67, and P53 are proliferation and cell cycle regulation markers that may predict tumor response to therapy. Aim: This study aimed to assess Top II, KI-67, and P53 expression and their effect on clinical outcome and response to therapy of non-muscle-invasive urothelial carcinoma. Materials and Methods: Fifty cases of non-muscle invasive urothelial carcinoma were collected; Top II, KI-67, and P53 expression was evaluated. Patients received treatment then tumor recurrence was correlated with the expression of previous markers. Results: There was a significant association between high Top II score, P53, and KI-67 and high tumor grade (P = 0.0001, 0.001, and 0.0001), submucosal infiltration (P = 0.0001 and 0.01), and recurrence (P = 0.01, 0.001, and 0.001). Conclusion: Top II, P53, and KI-67 may predict tumor response to therapy and the clinical outcome in non-muscle-invasive urothelial carcinoma.

Novel berberine derivatives: Design, synthesis, antimicrobial effects, and molecular docking studies / 中国天然药物

A series of berberine derivatives were synthesized by introducing substituted benzyl groups at C-9. All these synthesized compounds (4a-4m) were screened for their in vitro antibacterial activity against four Gram-positive bacteria and four Gram-negative bacteria and evaluated for their antifungal activity against three pathogenic fungal strains. All these compounds displayed good antibacterial and antifungal activities, compared to reference drugs including Ciprofloxacin and Fluconazole; Compounds 4f, 4g, and 4l showed the highest antibacterial and antifungal activities. Moreover, all the synthesized compounds were docked into topoisomerase II-DNA complex, which is a crucial drug target for the treatment of microbial infections. Docking results showed that H-bond, π-π stacked, π-cationic, and π-anionic interactions were responsible for the strong binding of the compounds with the target protein-DNA complex.

Novel berberine derivatives: Design, synthesis, antimicrobial effects, and molecular docking studies / 中国天然药物

A series of berberine derivatives were synthesized by introducing substituted benzyl groups at C-9. All these synthesized compounds (4a-4m) were screened for their in vitro antibacterial activity against four Gram-positive bacteria and four Gram-negative bacteria and evaluated for their antifungal activity against three pathogenic fungal strains. All these compounds displayed good antibacterial and antifungal activities, compared to reference drugs including Ciprofloxacin and Fluconazole; Compounds 4f, 4g, and 4l showed the highest antibacterial and antifungal activities. Moreover, all the synthesized compounds were docked into topoisomerase II-DNA complex, which is a crucial drug target for the treatment of microbial infections. Docking results showed that H-bond, π-π stacked, π-cationic, and π-anionic interactions were responsible for the strong binding of the compounds with the target protein-DNA complex.

Trimethoxy-benzaldehyde ciprofloxacin schiff base induces apoptosis in human hepatocarcinoma SMMC-7721 cells / 中国药学杂志

OBJECTIVE: To investigate the effect of 1-cyclopropyl-6-fluoro-7-(piperazin-1-yl)-3-[5-benzylsulfanyl-4-(3, 4, 5-trimethoxybenzylidene)amino-4H-1, 2, 4-triazol-3-yl]-quinolin-4(1H) -one (M27) on apoposis in hepatocarcinoma SMMC-7721 cells in vitro. METHODS: SMMC-7721 cells, colon adenocarcinoma cells(HCT-116) and leukemia cell line JURKET were treated by M27 with different concentrations for different time in vitro, the inhibitory effect of M27 and its precursor ciprofloxacin on the cell proliferation were examined by MTT assay. Cell apoptosis was determined by Hoechst 33258 fluorescence staining and TUNEL assay. The effect of M27 on topoisomerase II activity was measured using agarose gel electrophoresis by Plasmid pBR322 DNA as the substrate. Mitochondrial membrane potential(Δψm) was measured by high content screening imaging system. The p53, Caspase-9, Caspase-3, Caspase- 8, Bcl-2, Bax and cytochrome C protein expressions were determined by Western blotting analysis. RESULTS: The proliferation of the cancer cells was inhibited by M27 at 10-60 μmol·L-1 in time-and dose-dependent manner. Ciprofloxacin showed weak cytotoxicity against SMMC-7721 cell. SMMC-7721 cells treated by M27 with different concentrations for 24 h increased the percentage of apoptosis cells obviously (P < 0.05) with a decrease in the mitochondrial membrane potential. Compared with control group, M27 influenced obviously DNA topoisomerase II activity, stimulated DNA cleavage and inhibited DNA reunion mediated by topoisomerase II. In addition, M27 increased protein expression of p53, Bax, Caspase-8, Caspase-9, Caspase-3, as well as the cleaved activated forms of Caspase-9, Caspase-8 and Caspase-3 significantly, whereas the expression of Bcl-2 decreased. There was a significant increase of cytochrome C in the cytosol after 24 h of treatment with M27 and a decrease in the mitochondrial compartment. CONCLUSION: M27 as a fluoroquinolone derivative exerted potent anticancer activity through the mechanism of eukaryotic topoisomerase II poisoning. The growth inhibition is mainly mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

Proposal of Dual Inhibitor Targeting ATPase Domains of Topoisomerase II and Heat Shock Protein 90

There is a conserved ATPase domain in topoisomerase II (topo II) and heat shock protein 90 (Hsp90) which belong to the GHKL (gyrase, Hsp90, histidine kinase, and MutL) family. The inhibitors that target each of topo II and Hsp90 are intensively studied as anti-cancer drugs since they play very important roles in cell proliferation and survival. Therefore the development of dual targeting anti-cancer drugs for topo II and Hsp90 is suggested to be a promising area. The topo II and Hsp90 inhibitors, known to bind to their ATP binding site, were searched. All the inhibitors investigated were docked to both topo II and Hsp90. Four candidate compounds as possible dual inhibitors were selected by analyzing the molecular docking study. The pharmacophore model of dual inhibitors for topo II and Hsp90 were generated and the design of novel dual inhibitor was proposed.

Multiplication of Chromosome 17 Centromere Is Associated with Prognosis in Patients with Invasive Breast Cancers Exhibiting Normal HER2 and TOP2A Status / 한국유방암학회지

PURPOSE: This study aimed to investigate the clinical significance of chromosome 17 centromere (CEP17) multiplication (increased copy number of CEP17) related to human epidermal growth factor receptor 2 (HER2) and topoisomerase II alpha (TOP2A) status in patients with invasive breast cancer. METHODS: We constructed tissue microarrays using 594 invasive breast cancer samples and performed single-color silver-enhanced in situ hybridization (SISH) assay for HER2, TOP2A, and CEP17 to assess for copy number aberrations. The association of CEP17 multiplication with patient survival was analyzed according to HER2 and TOP2A status. RESULTS: Among 567 informative cases, HER2 amplification was noted in 22.8%, TOP2A amplification in 8.3% and TOP2A deletion in 11.1%. CEP17 multiplication was identified in 33.2% and was significantly associated with worse overall survival (OS) (p=0.02) and disease-free survival (DFS) (p=0.02). CEP17 multiplication correlated with patient survival in patients with normal TOP2A or non-amplified HER2 status, but the prognostic significance was lost in those with altered TOP2A or amplified HER2. On multivariate analyses, CEP17 multiplication was an independent prognostic factor for poorer OS (p=0.02) and DFS (p=0.01) in patients with normal TOP2A and non-amplified HER2. CONCLUSION: CEP17 multiplication was identified as a promising prognostic marker in patients with invasive breast cancer exhibiting either non-amplified HER2 or normal TOP2A status.

Multiplication of Chromosome 17 Centromere Is Associated with Prognosis in Patients with Invasive Breast Cancers Exhibiting Normal HER2 and TOP2A Status / 한국유방암학회지

PURPOSE: This study aimed to investigate the clinical significance of chromosome 17 centromere (CEP17) multiplication (increased copy number of CEP17) related to human epidermal growth factor receptor 2 (HER2) and topoisomerase II alpha (TOP2A) status in patients with invasive breast cancer. METHODS: We constructed tissue microarrays using 594 invasive breast cancer samples and performed single-color silver-enhanced in situ hybridization (SISH) assay for HER2, TOP2A, and CEP17 to assess for copy number aberrations. The association of CEP17 multiplication with patient survival was analyzed according to HER2 and TOP2A status. RESULTS: Among 567 informative cases, HER2 amplification was noted in 22.8%, TOP2A amplification in 8.3% and TOP2A deletion in 11.1%. CEP17 multiplication was identified in 33.2% and was significantly associated with worse overall survival (OS) (p=0.02) and disease-free survival (DFS) (p=0.02). CEP17 multiplication correlated with patient survival in patients with normal TOP2A or non-amplified HER2 status, but the prognostic significance was lost in those with altered TOP2A or amplified HER2. On multivariate analyses, CEP17 multiplication was an independent prognostic factor for poorer OS (p=0.02) and DFS (p=0.01) in patients with normal TOP2A and non-amplified HER2. CONCLUSION: CEP17 multiplication was identified as a promising prognostic marker in patients with invasive breast cancer exhibiting either non-amplified HER2 or normal TOP2A status.

A Case of Therapy-related ALL with MLL Gene Rearrangement Following Treatment of Breast Cancer / 대한진단검사의학회지

ALL with MLL gene rearrangement secondary to chemotherapy has been rarely reported. We report a case of therapy-related ALL (t-ALL) with MLL gene rearrangement in a patient who had undergone treatment for breast cancer. A 60-yr-old woman with breast cancer underwent breast-conserving surgery followed by 6 cycles of adjuvant chemotherapy (cyclophosphamide, epirubicin, and fluorouracil) and radiation therapy (dose, 5,040 cGy to the left breast and a 1,000 cGy boost to the tumor bed). A follow-up examination performed 14 months after the chemotherapy revealed no evidence of breast malignancy. However, the patient's complete blood cell count indicated acute leukemia: white blood cell count, 174.1x10(9)/L with 88% blasts; Hb level, 12.5 g/dL; and platelet count, 103.0x10(9)/L. Examination of the bone marrow aspirate smear revealed a high percentage of blasts (85.1% of all nucleated cells); the blasts showed a pro-B immunophenotype and were positive for CD19, CD79a, HLA-DR, CD34, and terminal deoxynucleotidyl transferase (TdT). Cytogenetic and FISH analyses revealed t(4;11)(q21;q23) and MLL gene rearrangement, respectively. The patient received induction chemotherapy with cyclophosphamide, vincristine, doxorubicin, and dexamethasone and achieved complete remission. Following consolidation chemotherapy, she underwent allogenic peripheral blood stem cell transplantation and has been clinically stable. To our knowledge, this is the first reported case of t-ALL with MLL gene rearrangement following treatment of breast cancer in Korea.

Immunohistochemical Studies of Fascin, MMP-9 Overexpression, and Proliferating Index as Prognostic Factors in Cases of a Colon Adenocarcinoma

PURPOSE: The malignant conversion of epithelial cells involves alterations in the expression and the function of cell-matrix and cell-cell adhesive systems that enable a switch to a migratory phenotype in tumor invasion and metastasis. Here, the author studies the prevalence and the potential clinical significance of fascin and Matrix metalloproteinase-9 (MMP-9) expression in relation to the progression of colon adenocarcinoma and of tumor cell proliferation as measured by using the topoisomerase II-alpha (Topo II-alpha) index. METHODS: Relatively well-preserved paraffin-embedded tissues of 120 cases of colon adenocarcinomas were immunohistochemically stained for fascin, MMP-9, and Topo II-alpha expression. A reaction was determined as being positive when more than 10% of the cells were positive for fascin, and/or MMP-9. The Topo II-alpha index is defined as the positive number of tumor cells divided by the total number of tumor cells counted times 100. At least 1,000 cells were counted for this analysis. A chi-square test, by using Epi info 2000, for Fascin and/or MMP-9 and a two-sided test for the Topo II-alpha index were employed with a significance of P65 yr, P=0.028), tumor grading (P=0.009), and lymph node metastases (P=0.005). However, MMP-9 immunoreactivity was not statistically associated with age, gender, tumor stage, or lymph node metastases. Fascin expression was statistically associated with MMP-9 expression, especially for left colon adenocarcinomas (P=0.0032). Although the topo II-alpha proliferating index was associated with lymph node metastasis (P<0.01), this result was not statistically associated with Fascin or MMP-9 expression. CONCLUSION: Fascin expression may be closely linked with tumor grading and lymph node metastasis of more aggressive colon adenocarcinomas and partly associated with MMP-9 expression in tumor invasion. However, further studies of fascin expression as an independent prognostic factor are required for the determination of significant relationships with other clinicopathologic indices.

Alteraciones cromosómicas estructurales inducidas por bioflavonoides de la dieta en linfocitos de anemia de Fanconi / Structural Chromosomal Alterations Induced by Dietary Bioflavonoids in Fanconi Anemia Lymphocytes

Introducción. La anemia de Fanconi es una enfermedad genética con herencia autosómica recesiva caracterizada por aplasia medular, predisposición a leucemia mieloide aguda, tumores sólidos y aumento en la inestabilidad cromosómica. Este síndrome puede considerarse como modelo biológico para analizar sustancias naturales con posible efecto genotóxico, difíciles de evaluar en células normales. Los objetivos de este estudio son describir y cuantificar las alteraciones cromosómicas estructurales inducidas por cinco flavonas, dos isoflavonas y una droga quimioterapéutica inhibidora de la topoisomerasa II, en cultivos de linfocitos de anemia de Fanconi a fin de determinar si existe un incremento en el número y tipo de daño cromosómico respecto al control. Materiales y métodos. Se analizaron los cromosomas de linfocitos estimulados con fitohemaglutinina M de una paciente con anemia de Fanconi. Se evaluaron 100 metafases de cada uno de los cultivos expuestos a las sustancias y control basal. Las alteraciones cromosómicas fueron documentadas con fotografías convencionales y digitales mediante analizador de imágenes. Se utilizó la prueba de c2 para determinar diferencias significativas entre el daño cromosómico entre cultivos expuestos y no expuestos con un valor de P <0,05. Resultados. Se encontraron 431 alteraciones cromosómicas en 1000 metafases analizadas; la genisteína tuvo el mayor efecto genotóxico, seguida de la genistina, fisetina, kaenferol, quercetina, baicaleína y miricetina. Las anomalías más frecuentes fueron las rupturas cromatídicas, rupturas cromosómicas, brechas (gaps) cromatídicas y cromosómicas, intercambios cuadri-radiales, cromosomas dicéntricos y rearreglos complejos. Conclusión. Los bioflavonoides genisteína, genistina y fisetina, presentes comúnmente en la dieta, aumentaron el número de alteraciones cromosómicas de manera significativa respecto al control en linfocitos de anemia de Fanconi.

Introduction. Fanconi anemia is an autosomal recessive disease characterized by a variety of congenital abnormalities, progressive bone marrow failure, increased chromosomal instability and higher risk to acute myeloid leukemia, solid tumors. This entity can be considered an appropriate biological model to analyze natural substances with possible genotoxic effect. The aims of this study were to describe and quantify structural chromosomal aberrations induced by 5 flavones,² isoflavones and a topoisomerase II chemotherapeutic inhibitor in Fanconi anemia lymphocytes in order to determine chromosomal numbers changes and/ or type of chromosomal damage. Materials and methods. Chromosomes stimulated by phytohaemagglutinin M, from Fanconi anemia lymphocytes, were analysed by conventional cytogenetic culture. For each chemical substance and controls, one hundred metaphases were evaluated. Chromosomal alterations were documented by photography and imaging analyzer. To statistical analysis was used chi square test to identify significant differences between frequencies of chromosomal damage of basal and exposed cell cultured a P value less than 0.05. Results. There were 431 chromosomal alterations in 1000 metaphases analysed; genistein was the more genotoxic bioflavonoid, followed in descendent order by genistin, fisetin, kaempferol, quercetin, baicalein and miricetin. Chromosomal aberrations observed were: chromatid breaks, chromosomal breaks, cromatid and chromosomal gaps, quadriratials exchanges, dicentrics chromosome and complex rearrangements. Conclusion. Bioflavonoids as genistein, genistin and fisetin, which are commonly present in the human diet, showed statistical significance in the number of chromosomal aberrations in Fanconi anemia lymphocytes, regarding the basal damage.

Amplification of the HER-2/neu Oncogene and Topoisomerase II-alpha by Chromogenic in Situ Hybridization in Breast Cancer

PURPOSE: HER-2/neu is the most frequently amplified oncogene in breast cancer. Topoisomerase II-alpha is a key enzyme in DNA replication and it is a molecular target for many anti-cancer drugs that are called topo II inhibitors; in addition, it is a new marker of proliferation. Because of the physical proximity of the ER-2/neu and topoisomerase II-alpha genes, co-amplification of the HER-2/neu and topoisomerase II-alpha may be important determinates of the response to chemotherapy for advanced breast cancer patients. METHODS: We studied the correlation of gene amplification of HER-2/neu and topoisomerase II-alpha by chromogenic in situ hybridization (CISH) in 43 infiltrating duct carcinomas of the breast. The over-expression of HER-2/neu protein and the staining index for the proliferation marker of topoisomerase II-alpha were examined immunohistochemically. The correlations between the status of HER-2/neu and topoisomerase II-alpha and the other clinicopathologic variables such as tumor size, lymph node metastasis, TNM stage, histologic grade, nuclear grade, and the estrogen receptor and progesteron receptor were investigated. RESULTS: Of the 43 infiltrating ductal carcinomas, the amplifications of HER-2/neu and topoisomerase II-alpha by CISH were observed in 8 cases (18.6%) and 14 cases (32.6%), respectively. Amplification of HER-2/neu showed the statistically significant correlations with tumor size, histologic grade and the topoisomerase II-alpha staining index. Amplification of topoisomerase II-alpha showed statistically significant correlations with axillary lymph node metastasis, the stage, the nuclear grade and the estrogen receptor status. CONCLUSION: These data suggest that amplification of HER-2/neu oncogene and topoisomerase II-alpha by CISH may be valuable for determining the response to chemotherapy, and detection of HER-2/neu and topoisomerase II-alpha in tumor sections may have prognostic value in human breast cancer.

p53, Heat Shock Protein 70 and Topoisomerase II Expression in Gallbladder Carcinoma

BACKGROUND: The present study was designed to investigate the expression of p53, Heat Shock Protein 70 (HSP70), and Topoisomerase (Topo) II alpha in the preneoplastic lesions and carcinomas of the gallbladder (GB) and to assess the correlation between the expression of these proteins and the clinicopathologic parameters by performing immunohistochemistry. METHODS: The immunohistochemical expressions of p53, HSP70 and Topo II alpha were evaluated in 38 gallbladder carcinomas and 3 adenomas. Fifteen CIS(s) and 8 dysplasias that were located adjacent to invasive carcinomas were also studied. RESULTS: A p53 expression was identified in 22 (57.9%) of the 38 GB carcinomas, in 9 (64.3%) of 14 CISs, and in none of the 8 dysplasias and 3 adenomas. A HSP70 expression was found in 11 (29%) of the 38 carcinomas, in 11 (78.6%) of 14 CIS(s), and in 4 (57.2%) of 7 dysplasias. A Topo II alpha expression was present in 36 (94.7%) of the 38 carcinomas, in 13 (92.9%) of 14 CIS(s), in 7 (100%) of 7 dysplasias and in 3 (100%) of 3 adenomas. p53 overexpression was related to the T stage of the primary tumor, while HSP70 and Topo II alpha were not related to any of the clinicopathologic parameters. CONCLUSION: p53 may be involved in GB carcinogenesis and in the progression of cancer. p53 may be also helpful for making the differential diagnosis between dysplasia and CIS. A further large study is needed to better elucidate the roles of HSP70 and Topo II alpha in GB carcinogenesis.

Identification of Candida Species by Multiplex Polymerase Chain Reaction / 대한임상미생물학회지

BACKGROUND: Polymerase chain reacation (PCR)-based methods have been described for rapid detection and identification of Candida spp. Multiplex PCR assay was developed using internal transcribed spacers and topoisomerase II gene for the accurate identification of Candida species. METHODS: We designed Dual Specificity Oligo (DSO) primers for multiplex PCR. Multiplex PCR was followed by agarose gel electrophoresis to test 8 type strains (C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae, C. dubliniensis) and 96 clinical isolates (C. albicans 51 isolates, C. parapsilosis 10 isolates, C. glabrata 10 isolates, C. tropicalis 9 isolates, C. krusei 6 isolates, C. guilliermondii 5 isolates, C. lusitaniae 5 isolates) of Candida spp. RESULTS: With multiplex PCR using DSO primers, the eight Candida type strains each could be easily differentiated and all 96 clinical isolates were identified as the same species as were identified by the conventional method. CONCLUSION: Multiplex PCR followed by electrophoresis can be useful for the simple and rapid identification of Candida species in routine laboratories.

Immunohistochemical Study of Metallothionein Expression in Colonic Adenocarcinoma: Correlation with p53, Topoisomerase II-alpha Expression and Apoptosis

PURPOSE: Although immunohistochemically detectable metallothionein (MT) overexpression has been described in proliferation epithelial tumor cells, the clinical significance of the expression remains to be elucidated. Therefore, the present article is focused on evaluating the possible significance of MT expression in colonic adenocarcinoma and its relationship with p53 overexpression, Topoisomerase II-alpha as new cell proliferating marker and apoptosis. METHODS: The following formalin-fixed paraffin embedded surgical or biopsied samples were immunohistochemically stained for MT, p53 and topoisomerase II-alpha, and performed in situ TUNEL method for evaluation of apoptotic cell ; normal control mucosa (78 cases), tubular adenomas (20 cases) and adenocarcinomas with various degree of differentiation (78 cases). RESULTS: The MT immunohistochmical reactivity was decreased in colonic adenocarcinoma than that of normal glandular epithelial and tubular adenoma, with the frequency of MT expression in colonic adenocarcinoma depending upon tumor differentiation only. But the frequency of p53 expression was correlated with T-stage, lymph node metastasis and clinical staging, while topoisomerase II-alpha expression and apoptosis in colonic adenocarcinoma were correlated with lymph node metastasis and clinical staging. The immunohistochemical expression of MT and p53 expression in colonic adenocarcinoma was inversely correlated. Also, the inverse correlation between MT expression and expression of toposiomerase II-alpha indices and apoptotic indices were noted. CONCLUSION: These data suggest that MT expression may play a role in proliferative activity and apoptosis in colonic adenocarcinoma. Although MT expression is correlated to tumor differentiation, further studies of a possibility of prognostic factor, such as p53, are required for the determination of significant relationships in other clinicinopathologic indices.

Proliferative Markers of Breast Cancer / 한국유방암학회지

PURPOSE: The mitotic index (MI) and Ki-67 labeling index have been used as cell proliferative markers in the various tumors. Topoisomerase II alpha (Topo II alpha) is also expressed in proliferating cells. The aim of this study was to evaluate the correlations between the MI, Ki-67, and Topo II alpha expression as proliferative markers of breast cancer. METHODS: The cell proliferative activity of 181 breast cancers was measured using MI, Ki-67 labeling index and Topo II alpha expression. The correlation between the measured markers was also analyzed. RESULTS: The MI, Ki-67, and Topo II alpha were significantly correlated with each other(p < 0.000). The MI and Ki-67 labeling index were associated with high histological grade, and absence of hormone receptor (p < 0.000). Topo II alpha expression was correlated with high histological grade (p < 0.000), absence of hormone receptor and HER-2/neu overexpression (p < 0.043). The MI, Ki-67, and Topo II alpha were not associated with any other clinical variables, such as age, tumor size, and lymph node status. CONCLUSION: The three proliferative indices were significantly associated with aggressive features of breast cancer, and significantly correlated with each other.

Correlation of Immunohistochemical Expression of MDR1, MRP1, Topoisomerase IIalpha with Prognostic Factors and Histoculture Drug Response Assay (HDRA) Result in Breast Carcinoma / 한국유방암학회지

PURPOSE: Drug resistance plays an important role in the failure of chemotherapy in breast cancer. The purpose of the study was to investigate the chemosensitive and chemoresistance indices of breast carcinomas and see if the in vitro chemosensitivity test correlated with the prognostic indices. METHODS: The immunohistochemical expressions of MDR1, MRP1 and topoisomerase IIalpha(topo IIalpha) were studied and then correlated these with the in vitro chemosensitivities using the histoculture drug response assay (HDRA) and clinicopathological factors in 51 breast carcinomas. RESULTS: In the breast carcinomas examined, the immunohistochemical expressions of MDR1, MRP1 and topo II alpha were strongly observed in 26 (51.0%), 16 (32.0%), 15 (31.3%) carcinomas, respectively. The MRP1 was more frequently expressed in poorly differentiated carcinomas (P= 0.006), and those of MDR1 and topo II alpha were more frequently observed in tumor overexpressing cerbB2 (P=0.038, P=0.036). The expression of MDR1 was related to that of topo II alpha (P=0.015). Comparing these markers with the in vitro chemosensitivities to cyclophosphamide, 5-FU, adriamycin, taxol and taxotere, no correlations were found between the expression of MDR1, MRP1, and topo II alpha but from the chemosensitivity using the HDRA, the growth inhibition rate for cyclophosphamide was higher in MRP1 expressing carcinomas (P=0.009). CONCLUSION: MDR1, MRP1 and topo II alpha were all found to be associated with the poor prognostic indices, but assessment of their immunohistochemical expressions did not allow for prediction of the response to chemotherapy by the in vitro chemosensitivity test in breast carcinomas.

The Immunohistochemical Expression of DNA Topoisomerase II-alpha and E2F-1 in the Breast Carcinoma and Their Relationship with Clinicopathologic Factors

PURPOSE: Clinical courses of breast cancer are very different, and concern for finding a predictable marker of breast carcinomas has increased. This study focused on the relationship between the expressions of DNA topoisomerase II-alpha as a proliferative marker, and E2F-1 as a transcription factor, with clinicopathological factors of infiltrating duct carcinomas of the breast. METHODS: We investigated the expressions of E2F-1 and DNA topoisomerase II-alpha in 43 patients with infiltrating ductal carcinomas using immunohistochemical staining, and the results were analyzed with regard to clinicopathological parameters. RESULTS: Among 43 infiltrating ductal carcinomas, 24 (55.8%) were immunohistochemically negative on E2F-1 and 19 (44.2%) were positive. The expression of E2F-1 correlated with increased tumor size, positive axillary lymph node meta stasis and high stage. The topoisomerase II-alpha index correlated with increased tumor size, positive lymph node metastasis, high stage, high histological grade and negative estrogen receptor. The expression of E2F-1 and the topo II-alpha index were significantly correlated. CONCLUSION: These results suggest that the expressions of DNA topoisomerase II-alpha and E2F-1 play some role as prog nostic factors for infiltrating duct carcinomas of the breast, but much more study will be required.

The Immunohistochemical Expression of E2F-1 Protein and DNA Topoisomerase II-alpha E2F-1 Protein in Colorectal Cancer and Their Relationship with Clinicopathologic Factors

PURPOSE: E2F-1 is a transcriptor that converts G1 to S in the cell cycle, and Topoisomerase II-alpha is a key enzyme in the metabolism of DNA, and an indicator of cell replication. The purpose of this study was to evaluate the clinical validity of E2F-1 and Topoisomerase II-alpha as prognostic factors in colorectal cancer. METHODS: The expressions of E2F-1 and Topoisomerase II-alpha were studied immunohistochemically using tumor specimen sections fixed with formalin and paraffin-embedded for 84 cases of colorectal cancer. The correlation between E2F-1 and Topoisomerase II-alpha expressions, and their relationship with the clinicopathological factors, such as tumor differentiation, tumor invasion, lymph node metastasis and tumor stage were investigated. RESULTS: Of the 84 specimens, 43 (51.2%) were immunohistochemically negative for E2F-1, and 41 (48.8%) were positive. The expression of E2F-1 correlated with poor tumor differentiation, increased lymph node metastasis and high tumor stage. The expression of Topoisomerase II-alpha also correlated with poor tumor differentiation, increased lymph node metastasis and high tumor stage. The E2F-1 and Topoisomerase II-alpha expressions indices were significantly correlated. CONCLUSION: These results suggest that the expressions of E2F-1 and DNA Topoisomerase II-alpha may play a role as a prognostic factor for colorectal cancer, but further studies will be required for its comfirmation.

Expression of c-erbB-2 Oncoprotein and DNA Topoisomerase II-alpha in Premalignant Lesions and Invasive Squamous Cell Carcinomas of the Uterine Cervix / 대한산부인과학회잡지

OBJECTIVE: The relationship was studied between expression of c-erbB-2 oncoprotein and topoisomerase II-alpha as proliferating marker in precancerous lesions and invasive squamous carcinomas of the uterine cervix. METHODS: Total 81 formalin-fixed, paraffin-embedded sections of low-grade intrasquamous lesion (22 cases), high-grade intraepithelial lesions (42 cases) and invasive squamous cell carcinomas (17 cases) in the uterine cervix were stained by immunohistochemistry for expression of the c-erbB-2 oncoprotein and topoisomerase II-alpha. RESULTS: The expression of c-erbB-2 oncoprotein and staining index (mean+/-S.D) of topoisomerase II-alpha were statistically significant between precancerous lesions and invasive carcinoma. The expression of c-erbB-2 oncoprotein has correlation with staining index (mean+/-S.D) of topoisomerase II-alpha. CONCLUSION: There results suggest that the expression of c-erbB-2 protein has relationship with progression of squamous lesions and topoisomerase II-alpha is an useful proliferating marker in the uterine cervix. And, the expression of c-erbB-2 protein has correlation with expression of topoisomerase II-alpha.

Expression of Multidrug Resistance Genes, mdr1, mrp, Topo IIalpha and Topo IIbeta in Childhood Acute Lymphoblastic Leukemia / 대한소아혈액종양학회지

PURPOSE: Multidrug resistance in the treatment of cancer mediated by several drug resistant genes impedes the successful management of cancer. The authors investigated the expression of drug resistant genes, including multidrug resistance (mdr1), multidrug resistance-associated protein (mrp), topoisomerase IIalpha (Topo IIalpha), and topoisomerase IIbeta (Topo IIbeta) in patients with childhood acute lymphoblastic leukemia (ALL; n=24) and in normal controls (n=6). METHODS: The levels of their transcripts in the bone marrow mononuclear cells were compared by semiquantitative RT-PCR, comparing the optical density ratio of PCR products of target genes to that of beta2-microglobulin. RESULTS: The expression of all 4 genes were detected in ALL patients as well as in normal controls except for Topo IIalpha, which were not detected in controls. The expression levels of mdr1, mrp and Topo IIbeta in ALL patients were significantly higher than those of normal controls [(2.1+/-2.2 vs 0.9+/-0.3, P> 0.024), (0.5+/-0.2 vs 0.2+/-0.1, P=0.016), and (0.7+/-0.2 vs 0.3+/-0.1, P=0.016)], respectively. The expression level of mdr1 gene transcript was relatively higher than those of mrp and Topo IIbeta. However, there was no correlation between the expression of multidrug resistant genes and survival and/or relapse of ALL. CONCLUSION: Though multidrug resistance genes did not serve as an independent prognostic factor, they might be used for markers for disease progression or relapse in childhood ALL. A longitudinal study of those genes is necessary to elucidate the prognostic significance in childhood ALL.