Topoisomerase Ⅱα Gene as a Marker for Prognostic Prediction of Hepatocellular Carcinoma: A Bioinformatics Analysis / 中国医学科学杂志(英文版)

Objective To investigate the expression of topoisomeraseⅡα (TOP2α) in hepatocellular carcinoma (HCC) and its role in predicting prognosis of HCC patients. Methods We used HCC-related datasets in UALCAN, HCCDB, and cBioPortal databases to analyze the expression and mutation of TOP2α and its co-expressed genes in HCC tissues. GO function and KEGG pathway enrichment of TOP2α and its co-expressed genes were identified. The TIMER database was used to analyze infiltration levels of immune cells in HCC. The impacts of TOP2α and its co-expression genes and the infiltrated immune cells on the survival of HCC patients were assayed by Kaplan-Meier plotter analysis. Results TOP2α and its co-expression genes were highly expressed in HCC (P< 0.001) and detrimental to overall survival of HCC patients (P< 0.001). TOP2α and its co-expression genes were mainly involved in cell mitosis and proliferation, and cell cycle pathway (ID: hsa04110, P = 0.001945). TOP2α and its co-expression genes were mutated in HCC and the mutations were significantly detrimental to overall survival (P = 0.0247) and disease-free survival (P = 0.0265) of HCC patients. High TOP2α expression was positively correlated with the infiltration of B cell (r = 0.459, P< 0.01), CD8+ T cell (r = 0.312, P< 0.01), CD4+ T cell (r = 0.370, P< 0.01), macrophage (r = 0.459, P< 0.01), neutrophil (r = 0.405, P< 0.01), and dendritic cell (r = 0.473, P< 0.01) in HCC. The CD8+ T cell infiltration significantly prolonged the 3- and 5-year survival of HCC patients (all P< 0.05), and CD4+ T cell infiltration significantly shortened the 3-, 5-, and 10-year survival of HCC patients (all P< 0.05). ConclusionTOP2α may be an oncogene, which was associated with poor prognosis of HCC patients and could be used as a biomarker for the prognostic prediction of HCC.

Aspectos ecoepidemiológicos, detección natural e identificación molecular de Leishmania spp. en Lutzomyia reburra, Lutzomyia barrettoi majuscula y Lutzomyia trapidoi / Eco-epidemiological aspects, natural detection and molecular identification of Leishmania spp. in Lutzomyia reburra, Lutzomyia barrettoi majuscula and Lutzomyia trapidoi

Resumen Introducción. La provincia de Pichincha, Ecuador, es un área endémica de leishmaniasis cutánea, en donde se han determinado como vectores los flebotomíneos antropofílicos con infección natural por Leishmania spp. Sin embargo, no se ha evaluado el papel en la transmisión de las especies zoofílicas. Objetivo. Evaluar la infección natural por Leishmania en dos especies de flebotomíneos zoofílicos, Lutzomyia reburra y Lu. barrettoi majuscula, y en una antropofílica, Lu. trapidoi, así como la endofagia y la sinantropía de estas especies en el noroccidente de Pichincha. Materiales y métodos. Los flebotomíneos se recolectaron en trampas de luz CDC colocadas en diferentes hábitats y altitudes en sitios que son focos de leishmaniasis cutánea. La infección con Leishmania spp. se detectó en el ADN genómico de hembras de las especies de flebotomíneos de interés. Se amplificó el gen espaciador interno de la transcripción del ARN ribosómico, unidad I (ITS1), y los genes de las topoisomerasas mitocondrial II (mtTOPOII) y nuclear II (TopoII). Se determinaron los porcentajes de positividad para Leishmania a escala espaciotemporal, la proporción de endofagia y el índice de sinantropía. Resultados. Se determinó la presencia de infección natural por Le. amazonensis en Lu. reburra (9,5 %) y Lu. b. majuscula (23,8 %); en Lu. trapidoi se detectaron Le. amazonensis, Le. brazilienis y Le. naiffilainsoni. Los flebotomíneos eran asinantrópicos y con baja endofagia. Conclusión. Se registró por primera vez la presencia de infección natural en Lu. reburra y Lu. barrettoi majuscula por Le. amazonensis, y se demostró la importancia de los flebotomíneos zoofílicos en el mantenimiento del ciclo de transmisión de Leishmania spp. en focos endémicos.

Abstract Introduction: The province of Pichincha in Ecuador is an endemic area of cutaneous leishmaniasis, where anthropophilic sand flies with natural infection by Leishmania, have been reported as vectors. However, the role in transmission of zoophilic species has not been evaluated. Objective: To evaluate natural infection by Leishmania in two zoophilic phlebotomine sand fly species, Lutzomyia reburra and Lu. barrettoi majuscula, and one anthropophilic species, Lu. trapidoi, as well as the endophagy and synanthropism of these species in the northwest of Pichincha. Materials and methods: Phlebotomines were collected using CDC light traps in different habitats and altitudes with presence of cutaneous leishmaniasis. Leishmania infection was detected using genomic DNA from females of the collected sand flies. We amplified the internal transcribed spacer gene of ribosomal RNA I (ITS1), the mitochondrial topoisomerase II gene (mtTOPOII), and the nuclear topoisomerase II gene (TopoII). Percentages of positivity for Leishmania, at spatio-temporal scale, proportion of endophagy and synanthropism index were calculated. Results: Natural infection was determined for Le. amazonensis in Lu. reburra (9.5%) and Lu. b. majuscula (23.8%), while in Lu. trapidoi we detected Le. amazonensis, Le. brazilienis and Le. naiffilainsoni. Phlebotomines were asynanthropic and with low endophagy. Conclusion: Natural infection with Le. amazonensis was recorded for the first time in Lu. reburra and Lu. b. majuscula, demonstrating the importance of zoophilic phlebotomines in the maintenance of the Leishmania transmission cycle in endemic foci.

Estudo da regulação das DNA Estudo da regulação das DNA topoisomerases durante o desenvolvimento de resistência ao etoposídeo e do papel antitumoral de novos compostos em leucemias agudas / DNA topoisomerases's regulation study during resistance development toetoposide and antitumor role of new compounds in acute leukemias

Entre as neoplasias hematológicas, as leucemias agudas configuram o maior número de mortes a cada ano. A quimioterapia para estas neoplasias envolve inibidores de topoisomerase, como as antraciclinas, associados a outros fármacos. Entretanto, alguns pacientes não respondem ao tratamento devido ao desenvolvimento do fenótipo de resistência a múltiplas drogas (MDR), considerada a principal causa de refratariedade e falha no tratamento. Devido à alta taxa de proliferação celular, os tumores super expressam as DNA topoisomerases I e IIα humana (hTopo I e IIα), tornando essas enzimas bons alvos para o desenvolvimento de novos fármacos. Neste contexto, a procura por novos compostos capazes de aumentar a taxa de sobrevida global em pacientes com leucemias agudas e que sejam eficazes em células com fenótipo MDR se faz necessária. Os objetivos deste trabalho foram: a)desenvolver linhagens celulares de leucemias agudas resistentes ao etoposido (VP-16); b)caracterizar o fenótipo de resistência, mediado por alterações nas enzimas hTopo I e IIα; c)avaliar o mecanismo de ação dos novos compostos LQBs (LQB-118, -192, -223, -266, -268 e -326) e ácido pomólico (PA), como potenciais inibidores de hTopo I e/ou IIα; e d)investigar a atividade antitumoral dos compostos mais promissores nas linhagens de leucemias agudas resistentes em comparação às parentais. Foi demonstrado que dentre os compostos LQBs avaliados apenas LQB-118 e LQB-223 foram efetivos e específicos em inibir hTopoIIα. PA demonstrou ser um composto dual inibindo hTopo I e IIα. Os três compostos ativos não intercalam no DNA e atuam como inibidores catalíticos, não apresentando afinidade de interação ao sítio de ligação entre o DNA e camptotecina ou VP-16. Os estudos de modelagem molecular também sugeriram que LQB-118 e LQB-223 apresentam alta afinidade de ligação à região ATPase de hTopoIIα...

Acute leukemias represent the largest number of annual deaths from hematologic malignancy. The chemotherapy for these neoplasms involves topoisomerase inhibitors, such as anthracyclines, associated with other drugs. However, some patients do not respond to this treatment scheme because of the development of multiple drug resistance (MDR) phenotype.MDR phenotype is the main cause of refractoriness and treatment failure in acute leukemias. Due to the high rate of cell proliferation, tumors overexpress human DNA topoisomerases I and IIα (hTopo I and IIα), leading these enzymes as good targets for the development of new anticancer drugs. In this context, the searching for novel compounds capable of increase theoverall survival rate in patients with acute leukemias and be effective on cells with MDR phenotype is urgent. The objectives of this research are: a) todevelop acute leukemia cell linesresistant to etoposide (VP-16); b) to characterize the resistance phenotype mediated by changes on hTopo I and IIα; c) to evaluate themechanism of action of new compounds LQBs(LQB-118, -192, -223, -266, -268 and -326) and pomolic acid (PA), as potential inhibitors of hTopo I and/or IIα; and d) to investigate the antitumor activity of the most promising compounds on parental or resistant acute leukemia cell lines. It was demonstrated that among the LQBs evaluated only LQB-118 and LQB-223 were effective and specific to hTopo IIα and that PA inhibited both hTopo I and IIα. They did not intercalate into DNA and acted as catalytic inhibitors with poor affinity to interact with camptothecin or VP-16 biding site. The molecular modeling studies also suggested that LQB-118 and LQB-223 presented a high affinity to bindto ATPase region of hTopo IIα. Acute lymphoid CEM-R and myeloid leukemia U937-R cell lines were developed by exposition to increasing concentrations of VP-16...

A homeopathic nosode, Hepatitis C 30 demonstrates anticancer effect against liver cancer cells in vitro by modulating telomerase and topoisomerase II activities as also by promoting apoptosis via intrinsic mitochondrial pathway / 中西医结合学报

<p><b>OBJECTIVE</b>Homeopathic nosodes have seldom been scientifically validated for their anticancer effects. This study was conducted to examine if a recently developed hepatitis C nosode has demonstrable anticancer potential in cancer cells in vitro.</p><p><b>METHODS</b>Anticancer effects of Hepatitis C 30C (Hep C 30), if any, were initially tested on three cancer cell lines, HepG2 (liver cancer), MCF-7 (breast cancer) and A549 (lung cancer) and one normal liver cell line WRL-68 cells and subsequently a more thorough study using further scientific protocols was undertaken on HepG2 cells (against WRL-68 cells as the normal control) as HepG2 cells showed better anticancer response than the other two. Three doses, one at 50% lethal dose (LD50) and the other two below LD50, were used on HepG2 cells subsequently. Protocols like apoptosis induction and its possible signaling mechanism were deployed using immunoblots of relevant signal proteins and confocal microscopy, with particular reference to telomerase and topoisomerase II (Top II) activities, two strong cancer biomarkers for their direct relationship with divisional activities of cells and DNAs.</p><p><b>RESULTS</b>Hep C 30 induced apoptosis, caused distorted cell morphology typical of apoptotic cells, increased reactive oxygen species generation and produced increased DNA nicks. Further it enhanced pro-apototic signal proteins like Bax, cytochrome c and inhibited anti-apoptotic signal proteins, Bcl-2, cytochrome c and caspase-3, changed mitochondrial membrane potential and caused externalization of phosphatidylserine. The drug also decreased expression of two cancer biomarkers, Top II and telomerase, consistent with its anticancer effect.</p><p><b>CONCLUSION</b>Hep C 30 has demonstrable anticancer effects against liver cancer cells in vitro.</p>

Structural, mechanistic and functional insights into topoisomerase II / 生理学报

Topoisomerases are nuclear enzymes that regulate the overwinding or underwinding of DNA helix during replication, transcription, recombination, repair, and chromatin remodeling. These enzymes perform topological transformations by providing a transient DNA break, through which the unique problems of DNA entanglement that occur owing to unwinding and rewinding of the DNA helix can be resolved. In mammals, topoisomerases are classified into two types, type I topoisomerase (Top1) and type II topoisomerase (Top2), depending on the number of strands cut in one round of action. Top1 induces single-strand breaks in DNA, and Top2 induces double-strand breaks. In cells from vertebrate species, there are two forms of Top2, designated alpha and beta. Top2α is involved in the cellular proliferation and pluripotency, while Top2β plays key roles in neurodevelopment. In this review, we cover recent advances in structural, mechanistic and functional insights into Top2.

Histone acetylation modification of topoisomerase enzyme Ⅱα promoter regulation factors in patients with chronic benzene poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate histone acetylation modification of topoisomerase enzyme Ⅱα (TOPOⅡα) promoter regulation factors in patients with chronic benzene poisoning, to explore the possible regulatory mechanism of TOPOⅡα involved in toxicity of chronic benzene poisoning;</p><p><b>METHODS</b>The bone marrow samples were from 25 chronic benzene poisoning cases and 25 controls. The Chromatin Immunoprecipitation (ChIP) assay was carried out to study the possible mechanism of TOPOⅡα promoter regulation factors expression changes. TOPOⅡα promoter regulation factors mRNA were detected by RT-PCR technique.</p><p><b>RESULTS</b>(1) Compared with the control, the histone H4 acetylation, histone H3 acetylation level of TOPOⅡα promoter regulation factors SP1, ATF-2, SP3, NF-YA, P53, C-MYB, ICBP90, NF-M in chronic benzene poisoning patients decreased, with the significant difference (P<0.05) , except for C-JUN (P>0.05) ; (2) The mRNA expression of TOPOⅡαpromoter regulation factors SP1, NF-YA, C-MYB, C-JUN and NF-M were significantly lower than in the control with the significant difference (P<0.05) , while the expression of SP3、P53 mRNA increased (P<0.05) , ATF-2、ICBP90 mRNA wasn't changed (P>0.05) .</p><p><b>CONCLUSION</b>(1) Chronic benzene poisoning TOPO Ⅱα promoter regulation factors histone modification changes accompanied with mRNA level changed. (2) Histone acetylation modification of topoisomerase enzyme Ⅱα promoter regulation factors takes important role in the benezen's Hematopoietic toxicity.</p>

Expressions of HER2 and Topo IIα in breast cancer and its clinical significance / 中南大学学报(医学版)

To detect the expressions of human epidermal growth factor receptor 2 (HER2) and Topo IIα in breast cancer, and to analyze the clinical significance of neoadjuvant chemotherapy for the anthracycline-based drugs.
 Methods: The HER2 and Topo IIα gene and protein expressions in cancer tissues from 189 patients with breast cancer were detected by fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC). And the objective response rate (ORR) and pathological complete rate (pCR) were analyzed.
 Results: The HER2 protein expression in 46 patients (24.3%) and Topo IIα protein expression in 55 patients (29.1%) was 3+ by IHC or they were 49 (25.9%) and 94 (49.0%) by FISH, respectively. The ORR and pCR in HER2 negative or positive patients were 47.4% and 20.3% or 32.7% and 16.3%, respectively, with significant differences (All P<0.05). The ORR and pCR in Topo IIα positive or negative patients were 69.1% and 36.0% or 28.4% and 2.2%, respectively, with significant differences (All P<0.05).
 Conclusion: FISH and IHC were consistent in the determination of HER2 expression whereas they were inconsistent in the determination of Topo IIα expression. The amplification of Topo IIα can effectively improve the effect of the adjuvant treatment effect of the anthracyclines.

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.

International cooperation and health policy implementation in a post-conflict situation: the case of East Timor / Cooperação internacional e formulação de políticas de saúde em situação pós-conflito: o caso do Timor-Leste

This study centers on relationships among national and international actors in preparation of the first health policy document for East Timor, under the United Nations transitional administration, between 1999 and 2002. International cooperation support for the health system rehabilitation process during the post-conflict period is analyzed as part of reconstruction of the State in parallel with construction of the country's political and institutional framework. Knowledge, ideas, "ways of doing," and induced and accepted practices permeate an interplay of power relationships that condition both national political alliance-building and the architecture of international aid, pointing to input to a discussion of how these mechanisms interact at different conjunctures and times in different negotiating frameworks. .

Dedica-se, aqui, às relações entre diferentes atores na elaboração do primeiro documento de política de saúde para o Timor-Leste, sob a administração transitória das Nações Unidas, de 1999 a 2002. O apoio da cooperação internacional no processo de reabilitação do sistema de saúde no período pós-conflito é analisado como parte da reconstrução do Estado e concomitante à construção do arcabouço político e institucional no país. Conhecimentos, ideias, "modos de fazer" e práticas induzidas e aceitas entremeiam um jogo de relações de poder que condiciona tanto a articulação política nacional quanto a arquitetura da ajuda externa, apontando elementos para a discussão de como esses mecanismos se organizam em conjunturas diferentes de negociação.

The quantification and significance of muscle segment homeobox gene Msx2, human topoisomerase II-α, HPV16 and VEGF in sinonasal inverted papilloma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the quantification and significance of Msx2, topoII-α; HPV16 and VEGF in sinonasal inverted papilloma(SNIP), to study the correlation among the four factors,and to discover the relationship between Msx2 and topoII-α in the process of SNIP malignant transfomation.@*METHOD@#Real-time quantitative Polymerase Chain Reaction (RT-qPCR) was used to detect the expression of Msx2, topoII-α, HPV16 and VEGF in 13 cases of sinonasal inverted papilloma (SNIP), 10 cases of sinonasal squamous cell carcinoma(NSCC) and 10 cases of inflammatory nasal polyp paraffin (INP)tissues. According to the pathology results SNIP were divided into mild dysplasia, moderate dysplasia and severe dysplasia. All the data were analysised by SPSS17. 0, P<0. 05 was refered to statistically significant difference.@*RESULT@#The mRNA level of Msx2, topoII-α, VEGF and HPV16 in SNIP, NSCC tissues were significantly higher than in the INP tissues (P<0. 05). The expression differences of Msx2, topoII-α, HPV16 and VEGF mRNA level in SNIP tissues which were divided into three groups according to their pathological results,were all statistically significantly different between any two of the three groups (P< 0. 05). Using Pearson correlation coefficient analysis,we found positive correlation between any two of the mRNA level of Msx2, topoII-α, VEGF and HPV16 (P<0. 05).@*CONCLUSION@#Msx2 and topoII-α may play an important role in the process of SNIP Malignant transformation,which may be new targets for gene therapy of SNIP and NSCC.

Suppression of the growth of subcutaneous transplanted human liver cancer and lung metastasis in nude mice treated by sorafenib combined with fluorouracil / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to explore the inhibitory effect of sorafenib and 5-Fu on transplanted human liver cancer in nude mice, and to investigate the synergistic effect and mechanism between sorafenib and 5-Fu.</p><p><b>METHODS</b>The nude mouse model of human liver cancer was made by transplantation of human highly metastatic liver cancer cell line HCCLM3 cells, and the tumor-bearing nude mice were treated with sorafenib, 5-Fu or both, respectively, and mock-treated tumor-bearing nude mice as negative control. To assess the anti-tumor effect of sorafenib and the synergistic effect of sorafenib combined with 5-Fu by measuring the tumor weight and number of lung metastases. Moreover, the expressions of phosphorylated extracellular signal-regulated kinase (p-ERK), P-glycoprotein (P-gp) and topoisomerase 2-alpha (Topo IIa) in the nude mice were assayed by immunocytochemistry and Western blot.</p><p><b>RESULTS</b>The tumor weights and numbers of lung metastases were: (2.7 ± 0.825) g and 12.714 ± 6.317 in the negative control group, (0.933 ± 0.333) g and 4.333 ± 3.983 in the sorafenib group, (0.786 ± 0.212) g and 5.429 ± 4.315 in the Sorafenib + 5-Fu combination group, and (2.438 ± 0.793) g and 10.429 ± 6.241 in the 5-Fu group. Statistically, the tumor weights and numbers of lung metastases in the sorafenib group and combination group were significantly decreased, compared with that in the control group (P < 0.05). There was no significant difference in the tumor weight and number of lung metastases between the sorafenib group and the combination treatment group (P > 0.05). The expression levels of p-ERK, P-gp and Topo IIa proteins in the tumors after normalization were: negative control (0.017 ± 0.010, 0.085 ± 0.012, 0.103 ± 0.093), sorafenib group (0.010 ± 0.008, 0.044 ± 0.020, 0.020 ± 0.018), combination group (0.011 ± 0.007, 0.043 ± 0.023, 0.062 ± 0.026), and 5-Fu group (0.018 ± 0.009, 0.063 ± 0.032, 0.065 ± 0.034), respectively. Statistically, the expression of p-ERK, P-gp and Topo IIa in the Sorafenib group was significantly reduced compared with that of the control group (P < 0.05), and there was no significant difference in the expression of p-ERK, P-gp and Topo IIa between the sorafenib group and the combination treatment group (P > 0.05).</p><p><b>CONCLUSIONS</b>Sorafenib can inhibit not only the tumor growth and lung metastsis in the nude mouse models, but also reduce the expression of multidrug resistance proteins P-gp and Topo IIa as well. There is no significant advantage for the sorafenib + 5-Fu combination treatment than Sorafenib alone in inhibiting the expression of p-ERK, P-gp and Topo IIa.</p>

Cytotoxicity and Structure-activity Relationships of Naphthyridine Derivatives in Human Cervical Cancer, Leukemia, and Prostate Cancer

Naphthyridine compounds are important, because they exhibit various biological activities including anticancer, antimicrobial, and anti-inflammatory activity. Some naphthyridines have antimitotic effects or demonstrate anticancer activity by inhibiting topoisomerase II. These compounds have been investigated as potential anticancer agents, and several compounds are now part of clinical trials. A series of naphthyridine derivatives were evaluated for their in vitro cytotoxic activities against human cervical cancer (HeLa), leukemia (HL-60), and prostate cancer (PC-3) cell lines using an MTT assay. Some compounds (14, 15, and 16) were more potent than colchicine against all three human cancer cell lines and compound (16) demonstrated potency with IC50 values of 0.7, 0.1, and 5.1 microM, respectively. Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were used for quantitative structure-activity relationship (QSAR) molecular modeling of these compounds. We obtained accurate and predictive three-dimensional QSAR (3D-QSAR) models as indicated by the high PLS parameters of the HeLa (q2, 0.857; r2, 0.984; r2pred, 0.966), HL-60 (q2, 0.777; r2, 0.937; r2pred, 0.913), and PC-3 (q2, 0.702; r2, 0.983; r2pred, 0.974) cell lines. The 3D-QSAR contour maps suggested that the C-1 NH and C-4 carbonyl group of the naphthyridine ring and the C-2 naphthyl ring were important for cytotoxicity in all three human cancer cell lines.

Effect of miRNA-106a expression on the prognosis of patients with esophageal squamous cell carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the expression of miRNA-106a gene in esophageal squamous cell carcinoma (ESCC) and its association with clinicopathological features and prognosis of ESCC patients.</p><p><b>METHODS</b>Real-time fluorescence quantitative polymerase chain reaction (PCR) assay was used to determine the expression of miRNA-106a gene in esophageal cancer tissue and corresponding normal mucosa of 81 cases. Immunohistochemical technique was applied to detect the expression of p53, human epidermal growth factor receptor 2 (HER-2), DNA topoisomerase II (Topo II) and multidrug resistance-associated protein (MRP). The association of miRNA-106a expression with clinicopathological features, expression of related proteins, and prognosis of the patients was analyzed.</p><p><b>RESULTS</b>Among the 81 cases, under-expression of miRNA-106a gene was found in 48 cases (59.3%), normal expression in 22 cases (27.2%), and overexpression in 11 cases (13.6%). The expression of miRNA-106 gene was significantly associated with lymph node metastasis, pathological stage, and nerve invasion (all P < 0.05), significantly associated with expression of p53 (P = 0.006), and not significantly associated with expressions of HER-2, Topo II and MRP proteins (all P > 0.05). The expression of miRNA-106a gene was also significantly associated with progression-free survival (PFS, P = 0.032), but not significantly with overall survival (OS, P = 0.486). The results of Cox multivariate regression analysis showed that the PFS of ESCC patients was significantly correlated with lymph node metastasis (P = 0.029), but not correlated with the age, gender, tumor length, T stage, degree of differentiation, nerve invasion, and miRNA-106a expression (all P > 0.05).</p><p><b>CONCLUSIONS</b>In esophageal squamous cell carcinomas, the miRNA-106a gene is under-expressed, with tumor suppressor function, and may be regarded as a biological marker to assess the prognosis in patients with esophageal squamous cell carcinoma.</p>

Antibacterial activity and mechanism of baicalein / 药学学报

Baicalein (BAI) is an effective bactericide. The antibacterial activity and mechanism experiments were carried out by determining conductivity and content of macromolecules of membrane penetrability, the oxidative respiratory metabolism and protein synthesis changes and the inhibition of DNA topoisomerase activities. Electrical conductivity and the number of large molecules of BAI increased 2.48% and 1.8%, respectively, than that of the control. However, the membrane integrity did not destroyed by BAI directly. With BAI treatment, inhibition rates of activities for SDH and MDH were 56.2% and 57.4%, respectively, demonstrating that BAI could inhibit cell respiratory. After treated with BAI for 20 h, the total soluble content of proteins decreased by 42.83%. Moreover, the activities of DNA topoisomerase I and II were inhibited completely by 0.2 mmol x L(-1) BAI. These results indicated that BAI had obvious antibacterial activity on Staphylococcus aureus. The mechanism is that it could affect bacterial membrane penetrability, inhibit protein synthesis and influence SDH, MDH and DNA topoisomerase I and II activities to exert its antibacterial functions.

Expression and significance of Msx2 and topo II-alpha in sinonasal inverted papilloma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression and significance of muscle segment homeobox2 (Msx2) and topo II-alpha in sinonasal inverted papilloma (SNIP), and the relationship in the process of malignant transformation of SNIP.@*METHOD@#Immunohistochemical method was used to detect the expression of Msx2 and topo II-alpha in 32 cases of SNIP, 30 cases of inflammatory nasal polyp (INP) and 30 cases of SNIP with carcinoma. According to the pathology results, SNIP were divided into mild atypical hyperplasia, moderate atypical hyperplasia and severe atypical hyperplasia.@*RESULT@#The mean optical density of Msx2 in SNIP and SNIP with carcinoma tissues were 0.2183 +/- 0.0598 and 0.2521 +/- 0.0761,which were significantly higher than 0.1878 +/- 0. 0372 in the INP tissue (P<0.05 or 0.01). The mean optical density of topo II-alpha in SNIP and SNIP with carcinoma tissues were 0.2303 +/- 0.0397 and 0.2666 +/- 0.0483, which were significantly higher than 0.1978 +/- 0.0388 in the NIP tissue (P<0.01). There were significant difference of Msx2 and topo II-alpha in SNIP between any two of the three groups divided according to pathological morphology (P<0.01 or 0.05). The expression of Msx2 and topo II-alpha in SNIP were positively correlated (P<0.05).@*CONCLUSION@#Msx2 and topo II-alpha may play an important role in the occurrence and development of SNIP. So it can be used as new therapeutic targets.

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.

Cinética celular na endometriose profunda infiltrativa de reto-sigmoide: estudo anátomo-clínico / Cell kinetics in deep infiltrating endometriosis of rectosigmoid: an anatomoclinical study

INTRODUÇÃO: A endometriose, uma doença benigna, tem características invasivas com potencial proliferativo. O desenvolvimento das lesões pode ocorrer em decorrência de crescimento celular glandular e/ou estromal ou de alterações na cinética celular. Cinética celular refere-se ao equilíbrio entre a morte celular, ou apoptose, e a proliferação celular, que pode ser avaliada pela expressão de fatores de crescimento como, por exemplo, a topoisomerase 2-alfa (TOP2A). Também influenciam a cinética celular oncoproteínas como p53 e c-erB2, conhecidas por interferir na apoptose, podendo resultar em oncogênese. OBJETIVOS: O objetivo principal deste estudo foi comparar a cinética celular da endometriose infiltrativa de retosigmoide com a do endométrio eutópico de pacientes sem endometriose. Para tanto, foi avaliada a expressão de apoptose e de TOP2A bem como das oncoproteínas p53 e c-erB2. MÉTODOS: Foram obtidas amostras de lesões de endometriose envolvendo o reto-sigmoide de 60 mulheres com a doença e amostras de endométrio eutópico de 20 mulheres sem endometriose. A expressão de TOP2A e das proteínas p53 e c-erB2 foram quantificadas por técnica imuno-histoquímica. Método TUNEL foi utilizado para analisar os padrões de apoptose, que resultaram em índice de apoptose (IA). Índice de proliferação celular (IP) foi determinado a partir do nível de expressão de TOP2A. Índice de renovação celular (IRC) foi calculado pela razão entre IP e IA. As análises imuno-histoquímicas foram realizadas tanto no tecido endometrial como um todo, quanto nos componentes estromal e glandular separadamente. Coeficiente de Correlação de Spearman foi aplicado para identificar eventuais correlações entre variáveis clínicas, morfológicas (tamanho, quantidade e nível de invasão das lesões) e experimentais. RESULTADOS: Na análise da amostra do tecido como um todo, não foram evidenciadas diferenças entre os grupos experimental e controle em relação ao IA (p = 0,389). Por outro lado, o IP foi...

BACKGROUND: Endometriosis, a benign disease, has invasive features with its proliferative potential. Development of lesions may occur due to stromal and/or glandular cell growth and to alterations in cellular kinetics. Cellular kinetics involves a balance between the regulation of cell death, or apoptosis, and cell growth, that can be evaluated by the expression of growth factors, such as topoisomerase 2- alpha (TOP2A). Oncoproteins, such as p53 and c-erB2, known to affect apoptosis resulting in oncogenesis, also influence cellular kinetics. OBJECTIVES: The main objective of this study was to compare the cellular kinetics in deep endometriosis involving the recto-sigmoid to eutopic endometrium from patients without endometriosis. Apoptosis and TOP2A expression were primarily evaluated, as well as p53 and c-erB2 expression. METHODS: Study samples were obtained from endometriosis lesions involving the recto-sigmoid in 60 women, and control samples were obtained from eutopic endometrium from 20 women without endometriosis. The expression of TOP-2A, p53 and c-erB2 proteins were quantified using immuno-histochemistry. TUNEL method was used in the analysis of apoptosis patterns, and the apoptosis index (AI) was derived. The proliferation index (PI) was derived from the level of expression of TOP-2A. Cellular renew index (CRI) was calculated from the ratio of the PI and AI. Immunohistochemical analyses were performed in two ways: on the tissue collectively, and on the stromal and glandular components separately. Spearmans correlation coefficient was used to identify the correlation between clinical, morphological (size, number and level of invasion of lesions) and the study variables. RESULTS: When looked at collectively, there was no difference in the AI between study and control groups (p = 0.389). PI, however, was noted to be significantly higher in the control samples (p < 0.001). When evaluating the stromal cells separately from the glandular components...

Correlation of TOP2A gene expression and survival of breast cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to assess the TOP2A RNA expression and the relationship of TOP2A protein expression with metastasis-free interval in breast cancer patients.</p><p><b>METHODS</b>TOP2A expression was analyzed prior to surgery in 86 patients. The level of TOP2A gene amplification was analyzed by fluorescence in situ hybridization (FISH), its RNA expression level with RT-PCR, and their correlation with TOP2A protein expression was assessed by immunohistochemistry (IHC). The correlation between RNA expression level and metastasis-free interval in breast cancer patients was also analyzed.</p><p><b>RESULTS</b>Aberrations (amplification or deletion) of TOP2A copy number was observed in 25.6% (22/86) of the cases. TOP2A protein expression was detected in 66.3% (57/86) of the samples. There was a significant correlation between the TOP2A RNA expression and protein expression (P < 0.001). TOP2A gene expression was significantly associated with the metastasis-free interval in the breast cancer patients (P = 0.001). There was no significant correlation between TOP2A gene amplification and TOP2A protein expression (P = 0.211).</p><p><b>CONCLUSIONS</b>TOP2A RNA level is an objective and reliable prognostic indicator in breast cancer.</p>

The synergistic antitumor effects of berberine alpha-hydroxy-beta-decanoylethyl sulfonate with hydroxycamptothecine and its effect on topoisomerase / 药学学报

Synergistic antitumor effects of HB (berberine alpha-hydroxy-beta-decanoylethyl sulfonate, houttuyn berberine) with HCPT (hydroxycamptothecine), and its correlative mechanism were studied in vitro. MTT assay was employed to determine the cytotoxicity of HB combined with HCPT in tumor cells culture in vitro, IC50 and combination index (CI value) were used to evaluate the synergistic effects. The supercoiled DNA relaxation mediated by topoisomerase I & II was measured by agarose gel electrophoresis assay, and influence of HB was detected. The results showed that HB could inhibit the proliferation of tumor cells (SGC-7901, SW1116 and SW480) in vitro, and the inhibition ratio was increased, IC50 was reduced when combining with HCPT. CI value of the two drugs was less than 1 in HepG2, SW480, SGC-7901 and SW1116 cells. The lowest value was 0.447, 0.626, 0.161 and 0.178 in these tumor cells, respectively, further indicating HB has synergistic action with HCPT on suppressing tumor proliferation. The agarose gel electrophoresis assay showed HB can inhibit topoisomerase I & II activity of SW480 cells at the concentration of 2.0-8.0 mg x L(-1). HCPT is a typical inhibitor of topoisomerase I , the synergistic action between HCPT and HB on suppressing tumor proliferation is perhaps related to the congenerous inhibition of topoisomerase.