Perfil imuno-histoquímico de marcadores associados à transição epitelial-mesenquimal em lesões odontogênicas epiteliais benignas / Immunohistochemical profile of markers associated with epithelial-mesenchymal transition in benign epithelial odontogenic lesions

Os cistos e tumores odontogênicos, lesões que acometem o complexo maxilomandibular, podem exibir comportamento clínico-biológico mais agressivo. E a transição epitelialmesenquimal (TEM), processo pelo qual as células epiteliais perdem propriedades fenotípicas e adquirem características de células mesenquimais, incluindo maior motilidade e capacidade de invasão, através da regulação de fatores centrais de transcrição e suas vias associadas, podem fazer parte de características associadas às lesões odontogênicas. Dessa forma, o presente trabalho buscou analisar e comparar a expressão imuno-histoquímica de proteínas (Zeb1, Ecaderina, N-caderina e vimentina) envolvidas no processo de TEM, em lesões odontogênicas epiteliais benignas. A amostra consistiu em 88 casos de lesões odontogênicas, das quais compreendem 28 casos de ameloblastoma (AB), 30 de ceratocisto odontogênico (CO) e 30 de cisto dentígero (CD). Todos os espécimes submetidos à técnica imuno-histoquímica foram avaliados por microscopia de luz, e submetidos à escolha aleatória de 5 (cinco) campos, os quais foram fotografados em um aumento de 400x. A avaliação da expressão de cada marcador, a partir da análise em seu compartimento celular específico, foi feita de forma semiquantitativa, através da multiplicação dos escores associados à porcentagem de células imunomarcadas pelos escores relacionados à intensidade da coloração, sendo feita uma média dos cinco campos e o resultado definido como baixa expressão ou alta expressão, conforme metodologia utilizada. As associações foram feitas através do teste de Qui-quadrado e as correlações através do teste de correlação de Spearman. O nível de significância foi estabelecido em 5% (p < 0,05). Os resultados mostraram um pico de prevalência entre a 2ª e 3ª décadas de vida, em todas as lesões estudadas, com um acometimento maior em região posterior de mandíbula, e os ABs foram as lesões de maiores tamanhos, com 65% medindo acima de 2,5cm. A imuno-histoquímica evidenciou baixa expressão de Zeb1 em epitélio odontogênico das lesões estudadas, alta expressão de E-caderina e N-caderina, e uma expressão intermediária de vimentina. Quando realizada a correlação entre os marcadores, observou-se nos casos de AB uma correlação positiva e moderada entre Zeb1 nuclear e E-caderina membranar, Zeb1 citoplasmática e E-caderina membranar e entre E-caderina e vimentina citoplasmáticas. Como também uma correlação positiva moderada, nos casos de CD, entre Zeb1 nuclear e vimentina citoplasmática, e entre Zeb1 e vimentina citoplasmáticas. Logo, podemos concluir que Zeb1 pode estar atuando indiretamente nas vias responsáveis pelo crescimento e características morfológicas dessas lesões estudadas. Além disso, a expressão diferencial de E-caderina, Ncaderina e vimentina demonstraram fazer parte de um processo de TEM parcial nas lesões odontogênicas epiteliais benignas estudadas (AU).

Odontogenic cysts and tumors, lesions that affect the maxillomandibular complex, may exhibit a more aggressive clinical-biological behavior. And the epithelial-mesenchymal transition (EMT), a process by which epithelial cells lose phenotypic properties and acquire characteristics of mesenchymal cells, including increased motility and invasiveness, through the regulation of central transcription factors and their associated pathways, may be part of characteristics associated with odontogenic lesions. Thus, the present work sought to analyze and compare the immunohistochemical expression of proteins (Zeb1, E-cadherin, N-cadherin and vimentin) involved in the MET process in benign epithelial odontogenic lesions. The sample consisted of 88 cases of odontogenic lesions, comprising 28 cases of ameloblastoma (AB), 30 of odontogenic keratocyst (CO) and 30 of dentigerous cyst (CD). All specimens submitted to the immunohistochemical technique were evaluated by light microscopy and submitted to the random choice of 5 (five) fields, which were photographed at a magnification of 400x. The evaluation of the expression of each marker, based on the analysis in its specific cellular compartment, was carried out in a semi-quantitative manner, through the multiplication of the scores associated with the percentage of immunostained cells by the scores related to the intensity of staining, with an average of the five fields and the result defined as low expression or high expression, according to the methodology used. The associations were made using the chi-square test and the correlations using the Spearman correlation test. The significance level was set at 5% (p < 0.05). The results showed a prevalence peak between the 2nd and 3rd decades of life, in all the lesions studied, with a greater involvement in the posterior region of the mandible, and the ABs were the largest lesions, with 65% measuring above 2, 5cm. Immunohistochemistry showed low expression of Zeb1 in the odontogenic epithelium of the lesions studied, high expression of E-cadherin, high expression of N-cadherin and an intermediate expression of vimentin. When the correlation between the markers was performed, a positive and moderate correlation was observed in the cases of AB between nuclear Zeb1 and membrane E-cadherin, cytoplasmic Zeb1 and membrane E-cadherin and between cytoplasmic E-cadherin and vimentin. As well as a moderate positive correlation, in CD cases, between nuclear Zeb1 and cytoplasmic vimentin, and between cytoplasmic Zeb1 and vimentin. Therefore, we can conclude that Zeb1 may be acting indirectly on the pathways responsible for the growth and morphological characteristics of these lesions studied. Furthermore, the differential expression of E-cadherin, N-cadherin and vimentin was shown to be part of a partial TEM process in the benign epithelial odontogenic lesions studied (AU).

Intervention Mechanism of Yishen Huayu Prescription on Glomerular Podocyte Injury in Diabetic Nephropathy Rats Based on EMT Regulated by Wnt/β-catenin Pathway / 中国实验方剂学杂志

Objective:To investigate the intervention mechanism of Yishen Huayu prescription on glomerular podocyte injury in diabetic nephropathy （DN） rats based on epithelialmesenchymal transition （EMT） regulated by Wnt/<italic>β</italic>-catenin pathway. Method:The 60 SD rats were divided into control group， model group， Wnt-C59 group （0.03 g·kg^-1 Wnt/<italic>β</italic>-catenin pathway inhibitor）， low-dose group （8 g·kg^-1）， medium-dose group （16 g·kg^-1） and high-dose group （32 g·kg^-1）. After 12 weeks， various indexes ， including general signs， serum creatinine （SCr）， blood urea nitrogen （BUN）， renal index， urinary protein， blood glucose， renal pathological changes， podocyte and expressions of glomerular basement membrane injury and podocyte injury related proteins ［nephrin， synaptopodin］， Wnt/<italic>β</italic>-catenin pathway related proteins （Wnt1， <italic>β</italic>-catenin）， podocyte EMT related protein ［<italic>α</italic>-smooth muscle actin （<italic>α</italic>-SMA）， E-cadherin］， were compared between groups. Result:Compared with the control group， the renal tissue in the model group showed significant pathological changes， including diffuse thickening of glomerular mesangial matrix and severe foot process fusion， and a significant increase in SCr， BUN， renal indexes， urinary protein， blood glucose， Wnt1， <italic>β</italic>-catenin， and <italic>α</italic>-SMA expression levels （<italic>P</italic><0.05） as well as a significant decrease in nephrin， synaptopodin and E-cadherin expression levels（<italic>P</italic><0.05）. Compared with model group， SCr， BUN， renal index， urinary protein， blood glucose， Wnt1， <italic>β</italic>-catenin， and <italic>α</italic>-SMA expression levels in each intervention group significantly decreased （<italic>P</italic><0.05）， while the expression levels of nephrin， synaptopodin and E-cadherin significantly increased （<italic>P</italic><0.05）. Among intervention groups， the improvement of above indexes in high-dose Yishen Huayu prescription group was the most obvious （<italic>P</italic><0.05）， which was similar to the effect in Wnt-C59 group. Conclusion:Yishen Huayu prescription prevents podocyte EMT by inhibiting Wnt/<italic>β</italic>-catenin pathway， thereby repairing glomerular podocyte injury in rats with diabetic nephropathy.

Effect of FOXK2 overexpression on malignant biological behaviors of ovarian cancer SK-OV-3 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of forkhead box transcription factor (FOXK2) overexpression on the proliferation, migration, invasion and adhesion of human ovarian cancer SK-OV-3 cells and its related molecular mechanism. Methods: The open reading frame (ORF) of FOXK2 was cloned into lentivirus expression vector, which was then enveloped in HEK293T cells and transfected into human ovarian cancerSK-OV-3cells.TheoverexpressionefficiencywasdetectedbyqPCRandWesternblotting.Theproliferation, migration, invasion and adhesion of SK-OV-3 cells were detected by CCK-8, Scratch-healing, Transwell and Cell adhesion assays respectively, and the expressions of epithelial-mesenchymal transition (EMT) markers were detected by qPCR. Results: The FOXK2 overexpression vector was constructed successfully and packaged into lentivirus, which was then transfected into SK-OV-3 cells. After transfection, the expression of FOXK2 was significantly increased (P<0.01); the proliferation, migration and invasion of SK-OV-3 cells were significantly reduced while the adhesion ability was significantly increased (P<0.05 or P<0.01); and the expression levels of E-cadherin and β-catenin were significantly increased while that of vimentin and fibronection were significantly decreased (all P<0.01). Conclusion: Overexpression of FOXK2 in SK-OV-3 cells leads to a significant decrease in proliferation, migration and invasion but increase in adhesion. The molecular mechanism may be related to the reversion of the EMT process in tumor cells, suggesting that FOXK2 may be a potential target for the diagnosis and treatment of ovarian cancer.

Effect of lncRNA MEG3 on the malignant biological behaviors of cervical cancer standard regimen for advanced colorectal cancer / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the regulatory effect of lncRNA maternal imprinting gene 3 (MEG3) on proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of cervical cancer cells via miR-9-5p/SOCS5 axis. Methods: A total of 20 pairs of cancer and para-cancerous tissue specimens resected from cervical cancer patients in Chongqing Hospital of Traditional Chinese Medicine from January 2017 to June 2019 were collected for this study. Using liposome transfection technology, pcDNA3.1-MEG3,si-MEG3, miR-9-5p mimics, miR-9-5p inhibitor and their control plasmids were transfected into cervical cancer HeLa and SiHa cells respectively to construct overexpression and silence cell model. qPCR was used to detect the expression levels of MEG3, miR-9-5p and SOCS5 in cervical cancer tissues and cell lines. CCK-8 method and Transwell chamber method were used to detect cell proliferation, migration and invasion ability. The expression levels of E-cadherin and vimentin in cells were detected by cellular immunofluorescence experiments. Target genes were predicted through the Online Bioinformatics TargetScan database. Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-9-5p and MEG3, SOCS5, respectively. Results: Compared with para-cancerous tissues and cervical epithelial HcerEpic cells, the expressions of MEG3 and SOCS5 were significantly down-regulated and the expression of miR-9-5p was significantly up-regulated in cervical cancer tissues and cell lines (all P<0.01). TargetScan database analysis and Dual luciferase reporter gene assay confirmed the targeting relationship between miR-9-5p and MEG3 or SOCS5. MEG3 and SOCS5 significantly inhibited while miR-9-5p significantly promoted cell proliferation, migration and invasion ability (all P<0.01). MEG3 and SOCS5 promoted E-cadherin expression and inhibited vimentin expression, while miR-9-5p inhibited E-cadherin expression and promoted vimentin expression (P<0.05 or P<0.01). Conclusion: lncRNA MEG3 regulates proliferation, migration, invasion and EMT of cervical cancer cells via miR-9-5p/SOCS5 axis.

Cucurbitacin B inhibits proliferation and invasion of osteosarcoma 143b cells and promotes apoptosis / 肿瘤

Objective: To investigate the effects of cucurbitacin B (CuB) on the proliferation, invasion and apoptosis of osteosarcoma 143B cells, to explore the related molecular mechanisms. Methods: 143B cells were treated with different concentrations of CuB. The proliferation ability of 143B cells was detected by crystal violet staining and colony formation assay. The invasion ability of 143B cells was detected by Transwell chamber assay. The apoptosis rate and cell cycle distribution were detected by flow cytometry (FCM) method. The expression levels of epithelial-mesenchymal transition (EMT)-related markers including Vimentin, Snail and matrix metalloproteinase-9 (MMP-9) mRNAs and proteins were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The expression levels of apoptosis-related proteins [Bad, cleaved-caspase 3, cleaved-polyadenosine diphosphate ribose polymerase (PARP) and cyclin B1] and protein kinase B (PKB, Akt) signal pathwayrelated proteins [Akt, phospho-Akt (p-Akt), phosphatase and tensin homolog deleted on chromosome ten (PTEN) and phospho-PTEN (p-PTEN)] were detected by Western blotting. Results: Compared with the untreated control group, the proliferation and invasion of 143B cells treated with different concentrations of CuB were significantly inhibited (all P < 0.05). After treatment with 30 or 40 nmol/L CuB, the apoptosis rate of 143B cells was increased (both P < 0.05), the cell cycle was blocked in G2/M phase (both P < 0.01). The expression levels of MMP-9, Vimentin and Snail mRNAs and proteins were remarkably down-regulated (all P < 0.05) in 143B cells treated with CuB. The expression levels of apoptosis-related Bad, cleavedcaspase 3 and cleaved-PARP proteins were remarkably up-regulated (all P < 0.05) in 143B cells treated with CuB, the expression of cell cycle-related cyclin B1 protein was down-regulated (all P < 0.01) in 143B cells treated with CuB. At the same time, CuB reduced the expression level of p-Akt protein and increased the expression level of p-PTEN protein (both P < 0.05). Conclusion: CuB can inhibit the proliferation and invasion of osteosarcoma 143B cells, and promote apoptosis. These effects may be related to Akt pathway impediment and EMT processing attenuation.

lncRNA SBF2-AS1 promotes epithelial-mesenchymal transition of cervical cancer cells via regulating miR-140-5p/VEGFAaxis / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of lncRNA SBF2-AS1 on epithelial-mesenchymal transition (EMT) of cervical cancer HeLa cell via regulating miR-140-5p/VEGFA (vascular endothelial growth factor A) axis. Methods: After cell culture and transfection, the cells were divided into 5 groups: NC group, miR-140-5p mimic group, miR-140-5p mimic+pcDNA-VEGFA group, si-lncRNA SBF2-AS1+pcDNA-VEGFA group and si-lncRNA SBF2-AS1+miR-140-5p mimic group. The expression level of lncRNA SBF2-AS1 in cervical cancer tissues and cell lines was detected by qPCR. The targeted relationship between lncRNA SBF2-AS1, miR-140-5p and VEGFA was confirmed by Dual luciferase reporter gene assay. The expression levels of VEGFA and EMT-related proteins N-cadherin, Vimentin and E-cadherin in HeLa cells were detected by WB. The invasion and migration of HeLa cells were detected by Transwell. Results: lncRNA SBF2-AS1 was highly expressed in cervical cancer tissues and cell lines (P<0.05 or P<0.01). Dual luciferase reporter gene assay confirmed that lncRNASBF2-AS1 targetedly combined with miR-140-5p and VEGFAwas a target gene of miR-140-5p (P< 0.05). Knockdown of lncRNA SBF2-AS1 inhibited invasion and migration as well as EMT of HeLa cells. Further experiment confirmed that lncRNA SBF2-AS1 up-regulated the expression level of VEGFA via miR-140-5p, thereby promoting invasion, migration and EMT of HeLa cells. Conclusion: lncRNASBF2-AS1 promotes EMT of HeLa cells via miR-140-5p/VEGFAaxis.

The application of detecting BCAR1 (+)-circulating tumor cells in postoperative monitoring of early stage lung adenocarcinoma / 中华胸心血管外科杂志

Objective To explore the clinical significance regarding monitoring circulating tumor cells in early stage lung adenocarcinoma.Methods From November 2015 to January 2018,48 patients with stage Ⅰ lung adenocarcinoma were included in the study.BCAR1 expression in CTCs in peripheral blood were detected by using CanPatrolTM and RNA in situ hybridization detection.Results Among the 48 cases,CTCs and BCAR1 (+)-CTCs were detected in 41 cases(85.4％) and 30 cases(62.5％),respectively.Number of BCAR1 (+)-CTCs seemed to be significantly positively related to that of CTCs.BCAR1 (+)-CTCs were more likely to appear in the M-CTCS and E&M-CTCS.BCAR1 (+)-CTCs remarkably increased in three relapsed cases.Furthermore,there were 19 stable cases who had postoperative CTCs data:(1) in 12 patients,either CTCs or BCAR1 (+)-CTCs were significantly reduced or remained stable;(2) in 7 cases,CTCs increased,but BCAR1 (+)-CTCs remained stable in 2 cases,reduced in 1 case,and the other 4 cases underwent close follow-up.Conclusion Evaluation of BCAR1 (+)-CTCs possibly can be contributive to prediction of early lung adenocarcinoma recurrence or metastasis.

Identification of a subtype of poorly differentiated invasive ductal carcinoma of the breast based on vimentin and e-cadherin expression / Identificação de um subtipo pouco diferenciado de carcinoma ductal invasivo de mama, baseado na expressão da vimentina e E-caderina

Abstract Objective The use of molecular markers can identify a subgroup of tumors with distinct recurrence patterns. The present study aimed to characterize the immunohistochemical expression of vimentin (VIM), of E-cadherin (CDH1), and of cytokeratin 5 (CK5) in patients with invasive ductal carcinomas (IDCs). Methods We have constructed a tissuemicroarray (TMA) from87 patients with IDC of the breast. Immunohistochemistry (IHC) was performed to study the expression of estrogen and progesterone receptors (ER and PgR), human epidermal growth factor receptor 2 (HER2), VIM, CDH1, CK5, and Ki67. The tumors were classified as luminal A and B (n = 39), HER2 enriched (n = 25), and triple-negative (TNBC) (n = 23), based on the IHC expression. Results We have observed that luminal A and B tumors lack the VIM+/CDH1-/low phenotype. This phenotype was observed in 16.5% of the HER2+ tumors and in 60% of the TNBC tumors (p = 0.0001). Out of a total of 20 TNBC tumors, the CK5 (basal-like marker) was positive in 11 of them. The VIM+/CDH1-/low phenotype was observed in 5 CK5+ TNBC tumors (45%) and in 7 out of 9 CK5- TNBC tumors (78%) (p = 0.02). The median Ki67 index in the VIM+/CDH1-/low tumors was 13.6 (range: 17.8-45.4) compared with 9.8 (range: 4.1-38.1) in other tumors (p = 0.0007). The presence of lymph nodemetastasis was less frequent in patients with VIM+/CDH1-/low tumors (23% versus 61%; X2 test; p = 0.01). Conclusion Our findings suggest that the expression of VIM and CDH1 can identify a subset of IDCs of the breast with a mesenchymal phenotype associated with poor prognosis, high-grade lesion, and high mitotic index.

Resumo Objetivo O uso de marcadores moleculares pode identificar subtipos tumorais com diferentes taxas de recidiva. O objetivo do presente estudo é caracterizar a expressão imunohistoquímica da vimentina (VIM), da E-caderina (CDH1) e de CK5 em pacientes com carcinoma ductal invasivo (CDI) da mama. Métodos Utilizamos uma matriz de amostras teciduais (TMA, na sigla em inglês) de 87 pacientes com CDI da mama. Para avaliar a expressão dos receptores de estrogênio (RE) e receptores de progesterona (RP), HER2, VIM, CDH1, CK5 e Ki67, utilizamos imunohistoquímica. Os tumores foram classificados como luminal A e B (n = 39), HER2+ (n = 25) e triplo negativo (TNBC) (n = 23). Resultados Foi observado que tumores luminais A e B não expressaram o fenótipo VIM+/CDH1-/low. Este fenótipo foi observado em 16,5% dos tumores HER2+ e em 60% dos tumores TNBC (p = 0,0001). Dos 20 tumores TNBC, a CK5 (marcador de tumor basalóide) foi super expressa em 11 amostras. O fenótipo VIM+/CDH1-/low foi observado em 5 tumores CK5+ TNBC (45%) e em 7 dos 9 tumores CK5- TNBC (78%) (p = 0,02). A expressão média de Ki67 nos tumores VIM+/CDH1-/low foi 13.6 (amplitude de 17,8 a 45,4) comparado com 9,8 (amplitude de 4,1 a 38,1) nos outros tumores (p = 0,0007). A presença demetástase linfonodal foimenor em tumores com fenótipo VIM+/CDH1-/low (23% contra 61%; teste X2; p = 0,01). Conclusão Nossos achados sugerem que a expressão de VIM e CDH1 pode identificar um subtipo de CDI da mama com fenótipo mesenquimal associado a pior prognóstico, lesões de alto grau e alto índice mitótico.

Caracterização do efeito anti-neoplásico do butirato em duas linhagens celulares de rato derivadas de tumores hepáticos com diferente ní­vel de diferenciação / Characterization of the anti-neoplastic effect of butyrate in two mouse cell lines derived from hepatic tumors with different levels of differentiation

O carcinoma hepatocelular (HCC) é uma neoplasia primária com mau prognóstico e alta taxa de recorrência. Estudos recentes demostram que o HCC pode ser classificado em três subtipos segundo o perfil molecular. Destes subtipos, o HCC pouco diferenciado apresenta pior prognostico. Neste sentido, torna-se de particular interesse o estudo de compostos com efeitos diferenciadores e citotóxicos nas células destas neoplasias pouco diferenciadas. O butirato, um ácido graxo de cadeia curta produzido pela fermentação microbiana da fibra alimentar no intestino, tem demonstrado atividade anti-neoplásica e capacidade moduladora da diferenciação celular em diversos tipos celulares, incluindo linhagens de HCC humano e células progenitoras hepáticas. Assim, objetivou-se neste estudo, caracterizar o efeito do butirato de sódio (NaBu) em duas linhagens de células neoplásicas de rato: uma pouco diferenciada (GP7TB) e a outra, uma linhagem derivada de um HCC diferenciado (JM-1). A linhagem GP7TB mostrou maior resistência ao NaBu (ED50= 7,7 mM) do que as células JM-1 (ED50= 5,2 mM). A redução na viabilidade celular após 72 h de tratamento com NaBu esteve relacionada com a diminuição na proliferação celular e no caso das células GP7TB, de um aumento na apoptose. O tratamento com NaBu induziu alterações morfológicas nas duas linhagens celulares, porém apenas nas células do tipo GP7TB, essas alterações sugerem um processo de diferenciação/transdiferenciação celular. O aumento na expressão de genes envolvidos no controle da pluripotência de células tronco, assim como de alguns marcadores de células tronco, sugere que o NaBu induziu uma reprogramação profunda das células GP7TB. Por outro lado, a redução na expressão de genes relacionados com migração e plasticidade celular assim como de proliferação celular apontam que estas células diminuíram seu potencial invasivo e a capacidade de autorenovação. Embora sejam necessárias análises adicionais para confirmar o efeito observado nos perfis de expressão gênica, os resultados deste estudo sugerem que o NaBu apresenta efeito antineoplásico por meio da redução da proliferação, aumento da apoptose e modulação da expressão de genes associados com a transição epitéliomesenquimal em células com características tronco tumorais

Hepatocellular carcinoma (HCC) is a primary neoplasia with poor prognosis and high recurrence rate. Recent evidence suggests that HCC can be classified in three different subtypes based on their molecular profile. Among these subtypes, the poorlydifferentiated HCC has the worst prognosis. Therefore, the study of compounds with pro-differentiating and cytotoxic effects on poorly-differentiated neoplastic cells represents a matter of primary concern. Butyrate which is a short-chain fatty acid produced by microbial fermentation in the intestine, has demonstrated anti-neoplastic activity and pro-differentiating potential in several cell types, including, human HCC cell lines and liver progenitor cells. In this study, we aimed to characterize the effect of sodium butyrate (NaBu) on two neoplastic cell lines derived from rats: a poorlydifferentiated cell line (GP7TB) and, a cell line derived from a well-differentiated HCC. GP7TB showed increased resistance to NaBu treatment (ED50= 7.7 mM) compared to JM-1 (ED50= 5.2 mM). The reduction in cell viability observed after 72 h of treatment was explained by a reduction in cell proliferation and, in the case of GP7TB, by increased levels of apoptosis. The NaBu treatment induced morphological alterations in both cell lines. However, only in the case of GP7TB cells, the alterations suggested a differentiation/transdifferentiation process. The up-regulation of genes involved in pluripotency and genes expressing stem cell markers indicated that NaBu triggered a deep reprogramming of GP7TB cells. Besides, a down-regulation in the expression of genes related with cell migration and plasticity suggested that these cells reduced their invasive potential and their self-renewal capacity. Additional analyses are necessary to confirm the observed effect on gene expression profiles. However, the results of this study suggest that NaBu exert anti-neoplastic effects through apoptosis, reduction of cell proliferation and downregulation of genes associated with epithelial-mesenchymal transition of cancer stem-like cells

Progress in EIF5A2 gene in malignant tumors / 肿瘤

Eukaryotic translation initiation factor 5A2 (EIF5A2) is a small universally conserved acidic protein classified in the EIF family. It has been found that EIF5A2 is widely over-expressed in various carcinomas, such as lung cancer, breast cancer and cervical cancer. EIF5A2 mainly promotes the occurrence and development of carcinoma by inducing epithelial-mesenchymal transition. A combination therapy with N1-guanyl-1, 7-diaminohep-tane (GC7), ciclopirox (CPX) and cytokines pharmacological agents can inhibit the activity of EIF5A2 through interfering the process of hypusine post-translational modification, and it is expected to be a good choice for clinical medication. Studies have shown that EIF5A2, as a carcinogenic gene, may be an effective molecular biomarker in the diagnosis and treatment of cancer. This review summarizes the progress in EIF5A2 in carcinoma.

Progress in research on regulation of tumor occurrence and development by forkhead box Q1 gene / 肿瘤

The transcription factor forkhead box Q1 (FOXQ1), a member of the forkhead box superfamily, is involved in various biological processes, and it also plays an important role in tumorigenesis and development. FOXQ1 protein activates the transcription by directly binding to the promoter of target genes or indirectly interacting with other transcription factors, and it affects initiation, progression, invasion and metastasis of tumor by promoting epithelial-mesenchymal transition (EMT) and activating multiple cellular signal transduction pathways. Previous studies have indicated that FOXQ 1 gene is significantly associated with the occurrence and development of a variety of tumors. The expression level of FOXQ 1 gene can be used as a prognostic indicator to judge the prognosis of various tumors. This review focuses on the possible mechanism of FOXQ 1 gene in tumorigenesis and development.

AGE/RAGE regulates the epithelial-mesenchymal transition and stemness maintenance of colorectal cancer cells / 肿瘤

Objective: To explore the effects of advanced glycation end products (AGEs) on the epithelialmesenchymal transition (EMT) and stemness maintenance of colorectal cancer HCT116 cells and its possible mechanism. Methods: The abilities of proliferation, migration, invasion, clone formation and sphere formation of HCT116 cells after treatment with 200 μg/mL AGEs and 200 μg/mL AGEs in combination with 50 μmol/L phosphatidylinositol-3 kinase (PI3K)-specific inhibitor LY294002 were detected by CCK-8, wound-healing assay, Transwell chamber assay, clone formation assay and sphere formation assay, respectively. After treatment with 200 μg/ mL AGEs and 200 μg/mL AGEs in combination with 50 μmol/L LY294002, respectively, the ability of invasion and the proportion of CD133plus signsphered HCT116 cells as well as the expressions of CD133, matrix metalloproteinase-2 (MMP-2) and MMP-9 in sphered HCT116 cells were detected by Transwell chamber assay, FCM and Western blotting, respectively. The expressions of the receptor of advanced glycation end products (RAGE), E-cadherin, N-cadherin, vimentin, Snail, PI3K, phosphorylated PI3K (p-PI3K), protein kinase B1 (PKB1, Akt1) and phosphorylated Akt1 (p-Akt1) in sphered HCT116 cells after treatment with different concentrations of AGEs, 200 μg/mL AGEs and 200 μg/mL AGEs in combination with 50 μmol/L LY294002 respectively were detected by Western blotting. Results: The abilities of migration, invasion, clone formation and sphere formation of HCT116 cells after treatment with 200 μg/mL AGEs were improved (P < 0.01, P < 0.05, P < 0.01, P < 0.01, P < 0.05), and these effects were inhibited by 50 μmol/L LY294002 (P < 0.05, P < 0.05, P < 0.01, P < 0.01, P < 0.05). After treatment with AGEs, the ability of invasion and the proportion of CD133plus sign sphered HCT116 cells were increased (P < 0.01, P < 0.05), and the expression levels of CD133, MMP-2 and MMP-9 were up-regulated (all P < 0.05), whereas these effects were opposite by 50 μmol/L LY294002 (P < 0.01, P < 0.05, all P < 0.05). The expression level of RAGE in HCT116 cells after treatment with different concentrations of AGEs was up-regulated (P < 0.05). After treatment with 200 μg/mL AGEs, the expression levels of p-PI3K, p-Akt1, N-cadherin, vimentin and Snail proteins were up-regulated (all P < 0.05), whereas the expression level of E-cadherin protein was down-regulated (P < 0.05); but these effects were opposite by 50 μmol/L LY294002 (all P < 0.05). Conclusion: AGE activates the RAGE signal pathway and improves the abilities of proliferation, migration, invasion, clone formation and stemness maintenance of colorectal cancer HCT116 cells, and this mechanism may be related to EMT mediated by AGE/RAGE/PI3K-Akt signal pathway.

The inhibitory effect of MIR-154-3p on metastasis of non-small cell lung cancer cell lines / 肿瘤

Objective: To investigate the expression of microRNA-154-3p (miR-154-3p) in non-small cell lung cancer (NSCLC) tissues and cell lines, and explore the role of miR-154-3p in metastasis process of NSCLC. Methods: The expression levels of miR-154-3p in NSCLC tissues and the corresponding paracarcinoma tissues, metastatic and non-metastatic NSCLC tissues, NSCLC cell lines (A549, H292, H1975, 95D and SPC-A-1) and human embryo lung WI38 cells were detected by realtime fluorogenic quantitative-PCR (RFQ-PCR). The miR-154-3p-negative control (miR-154-3p-NC, as a negative control) or miR-154-3p-mimics was transiently transfected into A549 and H292 cells by liposome. The effects of miR-154-3p on migration and invasion abilities of NSCLC A549 and H292 cells were determined by wound healing assay and Transwell invasion assay. The expressions of epithelial-mesenchymal transition (EMT)-related proteins E-cadherin, zonula occludens-1 (ZO-1) (epithelial marker), N-cadherin and vimentin (mesenchymal marker) induced by miR-154-3p overexpression were detected by Western blotting. The bioinformatic softwares were used to predict the potential targets of miR-1 54-3p, and the interested target genes were further validated by Dual-Luciferase Reporter Assay and Western blotting. Results: The expression levels of miR-154-3p were significantly lower in NSCLC tissues (P < 0.001), metastatic NSCLC tissues (P < 0.05) and NSCLC cell lines (P < 0.01) than those in their corresponding controls. Overexpression of miR-154-3p attenuated the migration and invasion abilities of both A549 and H292 cells (P < 0.05). The expressions of E-cadherin and ZO-1 proteins were up-regulated in A549 and H292 cells (all P < 0.05), whereas the vimentin expression in A549 cells and N-cadherin expression in H292 cells were down-regulated (both P < 0.05). Dual-Luciferase Reporter Assay and Western blotting results revealed that miR-154-3p could negatively regulate the expression of BMI1. Conclusion: MiR-154-3p is lowly expressed in NSCLC tissues and cell lines. The overexpression of miR-154-3p can inhibit the migration and invasion abilities of NSCLC cells and block the EMT progression. BMI1 may be one target gene of miR-154-3p.

Progress in renal tubulointerstitial fibrosis / 国际儿科学杂志

Renal tubulointerstitial fibrosis is the final common pathway in end-stage renal disease.Whereas molecular mechanisms underlying fibrogenesis are still not completely understood.To know the latest pathogenesis and treatment methods is of important significance to prevent disease progression and save life.

Effect of local aldosterone on renal epithelial-mesenchymal transition in diabetic nephropathy rats / 中华肾脏病杂志

Objective To explore the effect of aldosterone on renal epithelialmesenchymal transition in streptozocin(STZ)-induced diabetic nephropathy rats. Methods Wistar rats were intraperitoneally injected with STZ(60 mg/kg)for the preparation of diabetic model.After 4 weeks,the rats with urinary protein＞30 mg/d were regarded as successful diabetic nephropathy(n=16),and were randomly divided into diabetic nephropathy(DN group,n=8)and spironolactone group(SP group,n=8).Then eight healthy rats were selected randomly as control group(N group,n=8).SP group rats were treated with spironolactone 40 mg·kg-1·d-1,and N group and DN group rats were given equal water.After 8 weeks,rats were sacrificed to collect urine,blood plasma,kidney tissue for detection of 24 h urinary protein,creatinine and renal pathological changes.Aldosterone concentration in plasma and kidney tissue was detected by mdioimmunoassay;E-cadherin,α-SMA protein expression by immunohistochemistry,Western blotting; E-cadherin,α-SMA mRNA expression by RT-PCR. Results Compared with N group,serum creatinine, urinary protein excretion in the DN rats were significantly higher (P＜0.01,respectively), E-cadhefin protein and mRNA were significantly reduced (P＜0.01, respectively),α-SMA protein and mRNA expression was up-regulated (P＜0.01, respectively). Aldosterone level of kidney tissue in DN rats was increased obviously [(24.71±5.30) ng/g vs (16.38±2.85) ng/g, P＜0.01], which was positively correlated with urinary protein excretion, serum creatinine and α-SMA protein (r=0.737, 0.574, 0.688, P＜0.01, respectively), and negatively correlated with E-cadherin protein (r=-0.659, P＜0.O1). While no significant difference was found in serum aldosterone among three groups. Compared with DN rats, urinary protein excretion, serum creatinine were reduced (P＜0.01, respectively), E-cadherin protein and mRNA were increased (P＜0.01, respectively), α-SMA protein and mRNA expression were decreased (P ＜0.01, respectively) in SP group rats.Conclusions Local aldosterone involves in renal epithelial-mesenchymal transition in diabetic nephropathy rat. Spironolactone can block the effect of aldosterone and play a role in renal protection.