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1.
Biomed Res Int ; 2021: 4873678, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34337013

RESUMO

LIHC (liver hepatocellular carcinoma) mostly occurs in patients with chronic liver disease. It is primarily induced by a vicious cycle of liver injury, inflammation, and regeneration that usually last for decades. The G protein nucleolar 2 (GNL2), as a protein-encoding gene, is also known as NGP1, Nog2, Nug2, Ngp-1, and HUMAUANTIG. Few reports are shown towards the specific biological function of GNL2. Meanwhile, it is still unclear whether it is related to the pathogenesis of carcinoma up to date. Here, our study attempts to validate the role and function of GNL2 in LIHC via multiple databases and functional assays. After analysis of gene expression profile from The Cancer Genome Atlas (TCGA) database, GNL2 was largely heightened in LIHC, and its overexpression displayed a close relationship with different stages and poor prognosis of carcinoma. After enrichment analysis, the data revealed that the genes coexpressed with GNL2 probably participated in ribosome biosynthesis which was essential for unrestricted growth of carcinoma. Cell functional assays presented that GNL2 knockdown by siRNA in LIHC cells MHCC97-H and SMCC-7721 greatly reduced cell proliferation, migration, and invasion ability. All in all, these findings capitulated that GNL2 could be a promising treatment target and prognosis biomarker for LIHC.


Assuntos
Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/terapia , Proteínas de Ligação ao GTP/metabolismo , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/terapia , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação para Baixo/genética , Proteínas de Ligação ao GTP/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Ontologia Genética , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Prognóstico , Reprodutibilidade dos Testes , Transdução de Sinais/genética
2.
Infect Drug Resist ; 13: 1775-1780, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32606827

RESUMO

Introduction: Antimicrobial resistance (AMR) of Neisseria gonorrhoeae (N. gonorrhoeae) becomes a grave public health problem in the world. A strengthened Antimicrobial Resistance Surveillance Program is needed to track the trend of AMR development. However, the lack of a proper antimicrobial susceptibility test (AST) method is a barrier to expand the AMR surveillance in China. Traditional agar dilution (AD) method is laborious and E-test strips have no approval license for clinical use. Herein, a Chinese group modified the microdilution (MD) method for clinical ASTs. The objective of this study is to compare the MD method with the AD method for N. gonorrhoeae AST. Materials and Methods: A total of 166 clinical isolates were tested for antimicrobial susceptibility of ceftriaxone, spectinomycin, azithromycin, ciprofloxacin, tetracycline, and penicillin using MD and AD method simultaneously. Results of MD method were read manually or automatically. Rates of essential agreement (EA), category agreement (CA), minor error, and very major error were compared. Results: The total EAs (compared with results read manually) of penicillin, tetracycline, ciprofloxacin, spectinomycin, ceftriaxone, and azithromycin were 90.4%, 97.0%, 85.5%, 100.0%, 94%, and 72.3%; and CAs were 82.5%, 94.0%, 100%, 100%, 95.2%, and 94%, respectively. Conclusion: We conclude that the MD method might be an alternative for clinical AST of N. gonorrhoeae in China. In particular, MD method has the potency of accurate differentiation of isolates resistant to ceftriaxone or azithromycin, which were empirically recommended for gonococcal treatment, but its quality remained suboptimal, and further improvement is needed for clinical use.

3.
Int J Antimicrob Agents ; 54(6): 757-765, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31425792

RESUMO

The prevalence of Neisseria gonorrhoeae infections has increased rapidly since 2015 in China. Antimicrobial resistance and molecular mobilisation in N. gonorrhoeae are two important factors driving this increasing prevalence. This study explored changes in antimicrobial susceptibility and molecular characteristics of N. gonorrhoeae collected in Guangdong, China (2013-2017). A total of 704 isolates were collected in two cities in Guangdong. MICs of major antimicrobials were determined. Penicillinase-producing N. gonorrhoeae (PPNG) and tetracycline-resistant N. gonorrhoeae (TRNG) were characterised, and N. gonorrhoeae multiantigen sequence typing (NG-MAST) was performed. High resistance to penicillin (68.2%), tetracycline (85.7%) and ciprofloxacin (98.2%) was observed. Spectinomycin, ceftriaxone and azithromycin appeared effective, with susceptibilities of 100%, 96.4% and 90.7%, respectively. Resistance to penicillin decreased significantly from 78.4% to 73.6% and to azithromycin from 11.9% to 3.7%. Total prevalence of PPNG, TRNG and PPNG/TRNG was 25.4%, 33.1% and 13.4%, respectively. Rates of PPNG decreased significantly from 37.3% to 23.9%, TRNG from 50.0% to 31.3%, and PPNG/TRNG from 23.5% to 11.7%. However, the ratio of African-type PPNG increased significantly (18.4% to 64.1%) compared with decreasing Asian-type PPNG (81.6% to 33.3%), and the ratio of American-type TRNG increased significantly (0% to 13.7%) compared with decreasing Dutch-type TRNG (100% to 86.3%). A total of 271 sequence types (STs) were identified by NG-MAST from 380 isolates collected in 2013, 2014 and 2017, with 145 novel STs. African-type PPNG is increasing and replacing Asian-type, and novel STs have emerged. Gonococcal isolates with new genotypes might contribute to the rising gonorrhoea epidemic in this area.


Assuntos
Epidemias , Gonorreia/epidemiologia , Gonorreia/microbiologia , Neisseria gonorrhoeae/efeitos dos fármacos , Antibacterianos/farmacologia , China/epidemiologia , Farmacorresistência Bacteriana Múltipla , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Neisseria gonorrhoeae/genética , Prevalência , Fatores de Tempo
4.
J Glob Antimicrob Resist ; 16: 202-209, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30321622

RESUMO

OBJECTIVES: The aim of the study is to identify ceftriaxone resistance-related genes in Neisseria gonorrhoeae. METHODS: Differences in gene expression were compared between ceftriaxone-susceptible N. gonorrhoeae isolates [minimum inhibitory concentration (MIC)=0.002-0.004mg/L] and isolates with decreased ceftriaxone susceptibility (MIC=0.125-0.5mg/L) using RNA-Seq (RNA sequencing). RESULTS: Total RNA of 10 clinical isolates was used to make libraries and generated an average of 24.07Mb reads per sample; these were assembled into 1871 mRNA genes. Moreover, 21 differentially expressed genes (DEGs) were found between the N. gonorrhoeae isolates with susceptibility and decreased susceptibility to ceftriaxone with a fold change of ≥2 (P<0.05), among which 11 were upregulated and 10 were downregulated. Furthermore, all DEGs were verified by quantitative reverse transcription PCR (qRT-PCR), which detected 25 clinical isolates with decreased ceftriaxone susceptibility and 21 ceftriaxone-susceptible isolates. In addition, seven DEGs revealed relative expression levels by 2-ΔΔCt and showed a statistical significance (P≤0.05). Analysis of Gene Ontology (GO) terms and KEGG pathway for functional enrichment showed that six DEGs were related to the cellular component and one DEG was related to the biosynthesis of antibiotics, and these results might be related to ceftriaxone resistance. CONCLUSIONS: Examining ceftriaxone resistance-related genes in N. gonorrhoeae is necessary owing to the high morbidity and antimicrobial resistance of N. gonorrhoeae, especially its eventual resistance to third-generation extended-spectrum cephalosporins (cefixime and ceftriaxone). Moreover, this report provides a new direction for the study and control of ceftriaxone-resistant N. gonorrhoeae.


Assuntos
Antibacterianos/farmacologia , Ceftriaxona/farmacologia , Farmacorresistência Bacteriana/genética , Neisseria gonorrhoeae/genética , Expressão Gênica , Gonorreia/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Neisseria gonorrhoeae/efeitos dos fármacos , Reação em Cadeia da Polimerase , RNA-Seq
5.
Diagn Microbiol Infect Dis ; 92(4): 325-331, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30292397

RESUMO

A microdilution method for the antibiotic susceptibility testing of Neisseria gonorrhoeae was established and improved, and the antibiotic resistance of N. gonorrhoeae samples isolated from 8 cities of Guangdong in 2016 was determined. The improved microdilution method was compared with the agar dilution method recommend by the World Health Organization (WHO) Western Pacific Region by testing the susceptibility of 100 clinical N. gonorrhoeae isolates. The essential agreement (EA), categorical agreement (CA), very major error (VME), major error (ME), and minor error (MIE) levels of the two methods were analyzed; the acceptable performance rates were measured as follows: ≥90% for EA or CA, ≤3% for VME or ME, and ≤7% for MIE. The EA, CA, VME, ME, and MIE of each method for 7 antibiotics, penicillin, tetracycline, ciprofloxacin, spectinomycin, ceftriaxone, cefixime, and azithromycin, were 96%-100%, 94%-100%, 0%-3%, 0%-2%, and 0%-6%, respectively. The Wilcoxon signed-rank test results indicated 94%-100% agreement between the 2 methods after excluding off-scale values (P > 0.05). The susceptibility of 634 N. gonorrhoeae strains to the 7 antibiotics above were tested through the microdilution method. The resistant rates of the isolates against ciprofloxacin, tetracycline, penicillin, and azithromycin were 99.8%, 88.3%, 53.8%, and 11%, and the percentages of the isolates with decreased susceptibility to ceftriaxone (minimum inhibitory concentration [MIC] ≥0.125 µg/mL) and cefixime (MIC ≥0.25 µg/mL) were 2.1% and 12%, respectively, in Guangdong. Among 8 cities, Shenzhen had the highest rates of resistance against penicillin (77.8%) and decreased susceptibility against ceftriaxone (5.6%). Zhuhai had the highest rates of decreased susceptibility against cefixime (30.1%), and Jiangmen had the highest azithromycin-resistant isolates (16.8%). The findings from this study indicated that the improved microdilution method is an alternative for testing the antimicrobial susceptibility of N. gonorrhoeae. The resistance rates of N. gonorrhoeae against penicillin, tetracycline, and ciprofloxacin were high. While ceftriaxone, cefixime, and spectinomycin remained effective against N. gonorrhoeae, their effectiveness seemed to be decreasing over time. Azithromycin therapy requires timely susceptibility test results.


Assuntos
Antibacterianos/farmacologia , Gonorreia/microbiologia , Testes de Sensibilidade Microbiana , Neisseria gonorrhoeae/efeitos dos fármacos , Antibacterianos/uso terapêutico , China , Cidades , Farmacorresistência Bacteriana , Feminino , Gonorreia/diagnóstico , Gonorreia/epidemiologia , Humanos , Masculino , Testes de Sensibilidade Microbiana/métodos , Neisseria gonorrhoeae/isolamento & purificação
6.
AMB Express ; 7(1): 48, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28233287

RESUMO

BACKGROUND AND OBJECTIVE: Gonorrhea is a sexually transmitted disease caused by the bacterium Neisseria gonorrhoeae. Rapid detection is crucial for effective prevention and treatment. This study developed and tested a low-cost effective method for detecting N. gonorrhoeae, especially in developing countries. METHODS: DNA from a N. gonorrhoeae standard strain, as well as from 26 genital secretion samples of gonorrhea patients, were isolated and used for loop-mediated isothermal amplification (LAMP) assay, which was conducted using either an automatic real-time PCR analyzer or a water bath. The amplified porA pseudogene sequence was compared with the NCBI database and the LAMP results were compared with that of the traditional culture method for its sensitivity and specificity. RESULTS: LAMP was able to detect Neisseria DNA at a concentration as low as 1 pg/µL (1 × 103 CFU/mL cells). The LAMP assay results obtained using an automatic real-time PCR analyzer was similar to that of the water bath. Relative to traditional culture, the sensitivity and specificity of the LAMP assay were 94.7 and 85.7%, respectively. CONCLUSION: LAMP was sensitive and reliable for detecting the porA gene of N. gonorrhoeae. It could be used as a rapid, low cost, and effective method for detecting N. gonorrhoeae.

7.
PLoS One ; 11(7): e0159658, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27434536

RESUMO

BACKGROUND: Chlamydia trachomatis is one of the most prevalent bacterial sexually transmitted infection in China. Although C. trachomatis genotypes can be discriminated by outer membrane protein gene (ompA) sequencing, currently available methods have limited resolutions. This study used a high-resolution genotyping method, namely, multilocus variable number tandem-repeat analysis with ompA sequencing (MLVA)-ompA, to investigate the local epidemiology of C. trachomatis infections among men who have sex with men (MSM) and men who have sex with women (MSW) attending a sexually transmitted diseases (STD) clinic in Guangzhou, China. METHODS: Rectal specimens from MSM and urethral specimens from MSW were collected between January 2013 and July 2014 at the Guangdong Provincial Center STD clinic. The specimens were sent to the laboratory for analyses. All specimens that were tested positive for C. trachomatis by the commercial nucleic acid amplification tests were genotyped by MLVA-ompA. RESULTS: Fifty-one rectal specimens from MSM and 96 urethral specimens from MSW were identified with C. trachomatis. One hundred and forty-four of the 147 specimens were fully genotyped by MLVA-ompA. Rectal specimens from MSM were divided into four ompA genotypes and urethral specimens from MSW into nine genotypes. No mixed infections were found among all specimens. The most frequent genotypes were D, G, J, E and F. All specimens were further divided into 46 types after ompA genotyping was combined with MLVA. Genotypes D-8.7.1 and G-3.4a.3 were the most frequent among MSM, whereas genotypes D-3.4a.4, E-8.5.1, F-8.5.1, and J-3.4a.2 were the most frequent subtypes among MSW. The discriminatory index D was 0.90 for MLVA, 0.85 for ompA, and 0.95 for MLVA-ompA. CONCLUSIONS: The most prevalent MLVA-ompA genotypes were significantly different between MSM and MSW from Guangzhou, China. Moreover, MLVA-ompA represented a more favorable degree of discrimination than ompA and could be a reliable complement for ompA for the routine subtypes of C. trachomatis.


Assuntos
Proteínas da Membrana Bacteriana Externa/genética , Infecções por Chlamydia/epidemiologia , Chlamydia trachomatis/genética , Genótipo , Homossexualidade Masculina , Parceiros Sexuais , Adolescente , Adulto , China/epidemiologia , Infecções por Chlamydia/diagnóstico , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/classificação , Chlamydia trachomatis/isolamento & purificação , Feminino , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus , Filogenia , Reto/microbiologia , Sequências de Repetição em Tandem , Uretra/microbiologia
8.
BMC Infect Dis ; 15: 412, 2015 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-26453557

RESUMO

BACKGROUND: Gonococcal antimicrobial resistance is a global problem. Different resistance plasmids have emerged and spread among the isolates of Neisseria gonorrhoeae worldwide and in China. We conducted this study to monitor the plasmid-mediated penicillin and tetracycline resistance among N. gonorrhoeae isolates in Guangzhou from 2002 to 2012. METHODS: Consecutive isolates of N. gonorrhoeae were collected from outpatients with gonorrhea attending the STD clinic in Guangdong Provincial Centre for Skin Diseases and STIs Control and Prevention. Penicillinase-producing N. gonorrhoeae (PPNG) isolates were analyzed by the paper acidometric method. Plasmid-mediated resistance to tetracycline in N. gonorrhoeae (TRNG) isolates was screened by the agar plate dilution method. Plasmid types were determined for TRNG and PPNG isolates using polymerase chain reaction (PCR). Minimum inhibitory concentrations (MICs) to penicillin and tetracycline were detected by the agar plate dilution. RESULTS: Of 1378 consecutive N. gonorrhoeae isolates, 429 PPNG and 639 TRNG isolates were identified. The prevalence of PPNG, TRNG, and PPNG/TRNG increased from 18.3 to 47.1 % (χ (2) = 31.57, p < 0.001), from 29.4 to 52.1 % (χ (2) = 16.28, p < 0.001) and from 10.0 to 26.2 % (χ (2) = 10.46, p < 0.001) between 2002 and 2012, respectively. Genotyping of plasmids among PPNGs showed that the majority (93.7 %) of the isolates were the Asian type plasmids, while the African type plasmid emerged in 2008 and rapidly increased to 14.0 % in 2012 (χ (2) = 25.03, p < 0.001). For TRNGs, all 639 isolates carried the Dutch type plasmid. MICs of penicillin G and tetracycline persisted at high levels and the MIC90s were 32-fold higher than the resistant cutoff point over 11 years. The prevalence rates of penicillin- and tetracycline-resistant N. gonorrhoeae varied from 90.9 to 91.1 % and from 88.3 to 89.3 % during 2002 to 2012, respectively. CONCLUSIONS: Resistance to penicillin and tetracycline among N. gonorrhoeae isolates remained at high levels in Guangzhou. The Asian type PPNG continued to spread and Dutch type TRNG was still the dominant strain. The African type PPNG has emerged and is spreading rapidly.


Assuntos
Farmacorresistência Bacteriana/genética , Gonorreia/epidemiologia , Gonorreia/microbiologia , Neisseria gonorrhoeae/efeitos dos fármacos , Neisseria gonorrhoeae/genética , Penicilinas/farmacologia , Antibacterianos/farmacologia , China/epidemiologia , Gonorreia/tratamento farmacológico , Humanos , Testes de Sensibilidade Microbiana , Neisseria gonorrhoeae/isolamento & purificação , Penicilina G , Penicilinase/genética , Penicilinase/metabolismo , Plasmídeos , Reação em Cadeia da Polimerase , Resistência a Tetraciclina/genética , beta-Lactamases/genética
9.
Jpn J Infect Dis ; 67(4): 288-91, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25056075

RESUMO

Antibiotic susceptibility of Neisseria gonorrhoeae in Guangzhou during 2002-2011 showed that resistance to penicillin and ciprofloxacin was high, while ceftriaxone remained effective although there was a trend towards reduced sensitivity.


Assuntos
Antibacterianos/farmacologia , Gonorreia/microbiologia , Neisseria gonorrhoeae/efeitos dos fármacos , Distribuição de Qui-Quadrado , China/epidemiologia , Estudos de Coortes , Farmacorresistência Bacteriana , Gonorreia/epidemiologia , Humanos , Testes de Sensibilidade Microbiana
10.
Int J Radiat Oncol Biol Phys ; 88(4): 955-60, 2014 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-24606854

RESUMO

PURPOSE: Ataxia telangiectasia mutated (ATM) protein is important in the DNA damage response because it repairs radiation-induced damage in cancers. We examined the effect of microRNA-223 (miR-223), a regulator of ATM expression, on radiation sensitivity of cancer cells. METHODS AND MATERIALS: Human embryonic kidney 293 T (293T) cells were infected with pLL3.7-miR-223 plasmid to generate the pLL3.7-miR-223 and -empty virus (EV) lentivirus (miR-223 and EV). A dual luciferase assay in which the reporter contained wild-type 3' untranslated region (UTR) of ATM was performed. U87MG cells were infected with miR-223 or EV to establish the overexpressed stable cell lines (U87-223 or U87-EV, respectively). Cells were irradiated in vitro, and dose enhancement ratios at 2 Gy (DER2) were calculated. Hind legs of BALB/c athymic mice were injected with U87-223 or U87-EV cells; after 2 weeks, half of the tumors were irradiated. Tumor volumes were tracked for a total of 5 weeks. RESULTS: The dual luciferase reporter assay showed a significant reduction in luciferase activity of 293T cells cotransfected with miR-223 and the ATM 3'UTR compared to that in EV control. Overexpression of miR-223 in U87MG cells showed that ATM expression was significantly downregulated in the U87-223 cells compared to that in U87-EV (ATM/ß-actin mRNA 1.0 vs 1.5, P<.05). U87-223 cells were hypersensitive to radiation compared to U87-EV cells in vitro (DER2 = 1.32, P<.01). Mice injected with miR-223-expressing tumors had almost the same tumors after 3 weeks (1.5 cm(3) vs 1.7 cm(3)). However, irradiation significantly decreased tumor size in miR-223-expressing tumors compared to those in controls (0.033 cm(3) vs 0.829 cm(3)). CONCLUSIONS: miR-223 overexpression downregulates ATM expression and sensitizes U87 cells to radiation in vitro and in vivo. MicroRNA-223 may be a novel cancer-targeting therapy, although its cancer- and patient-specific roles are currently undefined.


Assuntos
Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , MicroRNAs/fisiologia , Tolerância a Radiação/fisiologia , Animais , Linhagem Celular Tumoral , Síndrome de Down , Genes Reporter , Xenoenxertos , Humanos , Luciferases/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus
11.
Mol Oncol ; 8(2): 366-77, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24388360

RESUMO

In our previous study, we identified 1241 loci with somatic copy number alterations in human hepatocellular carcinoma (HCC) using Affymetrix SNP 6.0 arrays, and a putative cancer gene SERPINA5 was uncovered in a novel chromosomal region with recurrent copy number loss at 14q31.1-32.13. The SERPINA5 was reported to be deregulated in renal, breast, prostate and ovarian cancers. However, the roles of SERPINA5 in cancer remain greatly elusive. In this study, we found that the DNA dosage and expression level of the SERPINA5 gene were significantly decreased in HCC by quantitative real-time PCR. Notably, the expression levels of SERPINA5 negatively correlated with malignant progression of HCC. The SERPINA5 gene was further observed to reduce in vitro and in vivo metastatic potential of HCC cells. Moreover, secreted SERPINA5 protein also could inhibit the metastatic ability of HCC cells. Finally, we discovered that one of the mechanisms explaining SERPINA5 inhibition of HCC metastasis is through direct interaction with fibronectin and disruption of the fibronectin-integrin signaling pathway. These findings highlight an important role of SERPINA5 in the regulation of migratory and metastatic potentials of HCC and suggest a potential application of SERPINA5 in cancer treatment.


Assuntos
Carcinoma Hepatocelular/metabolismo , Integrina beta1/metabolismo , Neoplasias Hepáticas/metabolismo , Proteínas de Neoplasias/metabolismo , Inibidor da Proteína C/metabolismo , Transdução de Sinais , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Fibronectinas/genética , Fibronectinas/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Integrina beta1/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Proteínas de Neoplasias/genética , Inibidor da Proteína C/genética
12.
Int J Mol Med ; 30(6): 1321-6, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23023935

RESUMO

In the present study, we constructed a lentivirus vector encoding the miR-29a precursor and established two stably infected cell lines, PLC-29a and 97L-29a. The overexpression of miR-29a was confirmed by TaqMan RT-PCR and significantly suppressed the growth of the hepatocellular carcinoma cell lines MHCC-97L and PLC. Dual-luciferase reporter assays indicated that the SPARC mRNA 3'UTR was directly targeted by miR-29a since the mutated 3'UTR was not affected. Silencing SPARC expression by RNAi knockdown resulted in a similar effect as miR-29a overexpression on hepatocellular carcinoma (HCC) cell growth regulation. Anti-miR-29a oligonucleotides (AMOs) upregulated the levels of SPARC in the HCC cells. The phosphorylation of AKT/mTOR downstream of SPARC was inhibited in miR-29a-overexpressing HCC cells. We further examined and compared the expression levels of miR-29a in HCC tissues and the corresponding nearby non-cancerous liver tissues of 110 patients with HCC by qRT-PCR, and significantly lower expression of miR-29a was observed in the tissues affected by HCC. Our findings demonstrate that the expression of miR-29a is important in the regulation of the SPARC-AKT pathway and HCC growth.


Assuntos
Carcinoma Hepatocelular/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/metabolismo , MicroRNAs/fisiologia , Proteínas Supressoras de Tumor/genética , Adulto , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Técnicas de Silenciamento de Genes , Humanos , Fígado/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Osteonectina , Fosforilação , Processamento de Proteína Pós-Traducional , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Regulação para Cima
13.
Jpn J Infect Dis ; 63(5): 342-5, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20859001

RESUMO

This study was designed to determine the prevalence and distribution of Chlamydia trachomatis genotypes from clinical specimens in Guangzhou, China, obtained in the period 2005-2008. One hundred and ninety-four urogenital C. trachomatis samples were collected from sexually transmitted disease clinic patients, and the VS1-VS2 of OmpA gene was amplified by nested PCR and sequenced using an ABI-prism 3730 sequencer. Clinical C. trachomatis strains were genotyped and analyzed for a mutation with respect to the reference VS1-VS2 sequence. VS1-VS2 fragments with 453 bp were amplified from 194 clinical samples. Upon alignment with the sequences of the reference strains, 189 strains with discernible sequences were typed into 9 genotypes, while 5 with ambiguous sequences were considered to be mixed-serovar samples. The most prevalent genotypes were E (50, 26%), F (46, 24%), J (35, 19%), and D (24, 13%). There was no significant difference in the distribution of any of the genotypes detected during the study period, except for genotype K (P<0.01). A total of 16 (8%, 16/189) genetic variants of the OmpA VS1-VS2 of the reference strains were identified. Mutations occurred frequently for genotypes D (2/24, 8%), E (6/50, 12%), F (2/46, 4%), G (1/8, 13%), H (1/12, 8%), and K (4/11, 36%), with most of these being sense mutations that may result in amino acid substitution. Sequencing the OmpA VS1-VS2 enabled the genotype and sequence variations within each genotype to be analyzed. Genotypes E, F, J, and D continued to dominate among urogenital C. trachomatis, whereas genotype K increased significantly in Guangzhou between 2005 and 2008.


Assuntos
Infecções por Chlamydia/epidemiologia , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/genética , Doenças Bacterianas Sexualmente Transmissíveis/epidemiologia , Doenças Bacterianas Sexualmente Transmissíveis/microbiologia , Instituições de Assistência Ambulatorial , Proteínas da Membrana Bacteriana Externa/genética , Distribuição de Qui-Quadrado , China/epidemiologia , Estudos Transversais , Análise Mutacional de DNA/métodos , Feminino , Genótipo , Humanos , Masculino , Mutação , Reação em Cadeia da Polimerase
14.
Acta Pharmacol Sin ; 29(4): 413-20, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18358086

RESUMO

AIM: The aim of the present study was to understand the role of retinoic acid (RA) in the development of isolated patent ductus arteriosus and the features of arterial duct-derived vascular smooth muscle cells (VSMC). METHODS: The VSMC were isolated, and the biological characteristics and the response to RA were investigated in the arterial duct, aorta, and pulmonary artery VSMC from 6 human embryonic samples. Western blotting, immunostaining, and cell-based ELISA were employed to analyze the proliferation regulation of VSMC. RESULTS: The VSMC from the arterial duct expressed proliferating cell nuclear antigen (PCNA) at a significantly lower rate than those from the aorta and pulmonary artery, but expressed a higher level of Bax and Bcl-2. The expression level of PCNA or Bcl-2 was associated with the embryonic age. The effects of RA on the VSMC from the arterial duct were quite different from those from the aorta and pulmonary artery. In arterial duct VSMC, RA stimulated PCNA expression, but such stimulation could be suppressed by CD2366, an antagonist of nuclear retinoid receptor activation. In aorta or pulmonary artery VSMC, the expression response of PCNA to RA was insignificant. The ratio of Bax/Bcl-2 decreased in arterial duct VSMC after RA treatment due to the significant inhibition of Bax expression. CONCLUSION: The VSMC from the arterial duct possessed distinct biological behaviors. RA might be important in the development of ductus arteriosus VSMC.


Assuntos
Canal Arterial/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacos , Tretinoína/farmacologia , Antígenos CD/farmacologia , Aorta/citologia , Aorta/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Canal Arterial/citologia , Ativação Enzimática/efeitos dos fármacos , Feto/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Artéria Pulmonar/citologia , Artéria Pulmonar/metabolismo , Proteína X Associada a bcl-2/metabolismo
15.
World J Gastroenterol ; 11(37): 5763-9, 2005 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-16270382

RESUMO

AIM: To investigate the effect of N-(4-hydrophenyl) retinamide (4-HPR), the derivative of retinoic acid, on inhibition of migration, invasion, cell growth, and induction of apoptosis in hepatocellular carcinoma cells (HCCs) and malignant melanoma cells. METHODS: 4-HPR was chemically synthesized. Cellular migration and invasion were assayed by Borden chamber experiment. Cell growth was assayed by MTT chromometry. Apoptosis effect was measured using Hoechst 32258 staining and flow cytometry. Gene transfection was performed with lipofectamine. RESULTS: We observed that the migration of HCC and melanoma cells was significantly suppressed by 4-HPR and the migration cells were reduced to 58+/-5.03 (control 201+/-27.2, P<0.05, n = 4) in SMMC 7721-k3 HCC, and to 254+/-25.04 (control 302+/-30.1, P<0.05, n = 4) in melanoma cells after 6-h incubation with 4-HPR. The invasion through reconstituted basement membrane was also significantly reduced by 4-HPR treatment to 11.2+/-3.3 in SMMC 7721-k3 HCC (control 27+/-13.1), and to 24.3+/-3.2 in melanoma cells (control 67.5+/-10.1, P<0.05, n = 3). Cell growth, especially in melanoma cells, was also significantly inhibited. Furthermore, 3 micromol/L of 4-HPR induced apoptosis in B16 melanoma cells (37.11+/-0.94%) more significantly than all-trans retinoic acid (P<0.05), but it failed to induce apoptosis in SMMC 7721-k3 HCC. The mechanism for 4-HPR-induced apoptosis was not clear, but we observed that 4-HPR could regulate p27(kip1), and overexpression of cerebroside sulfotransferase (CST) diminished the apoptosis induced by 4-HPR in melanoma cells. CONCLUSION: 4-HPR is a potent inhibitor of HCC migration and inducer of melanoma cell apoptosis. CST and p27(kip1) expression might be associated with 4-HPR-induced apoptosis.


Assuntos
Antineoplásicos/uso terapêutico , Carcinoma Hepatocelular/tratamento farmacológico , Fenretinida/uso terapêutico , Neoplasias Hepáticas/tratamento farmacológico , Melanoma/tratamento farmacológico , Animais , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/patologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Fenretinida/farmacologia , Humanos , Neoplasias Hepáticas/patologia , Melanoma/patologia , Camundongos , Camundongos Nus , Sulfotransferases/genética , Sulfotransferases/metabolismo
16.
Biochem Biophys Res Commun ; 332(4): 934-40, 2005 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-15921657

RESUMO

In this study, significantly higher expression of Gal3ST-2 (Gal: 3-O-sulfotransferase-2) and 3'-sulfated glycoconjugates were observed in highly metastastic cancer cells and in larynx cancer tissues with lymph node metastasis than those in lowly metastatic cancer cells and larynx cancer tissues without metastasis (P<0.01, n=42). These results indicated that there was a marked correlation between the expression of Gal3ST-2 and tumor metastasis potential. After RNAi transfection, a striking morphological change of SMMC7721 hepatoma cells from polygon to shuttle shape and significant decrease in adhesion to sL-selectin and HUVEC were observed. Interestingly, the expression of integrin subunit alphaV was markedly downregulated and 3'-sulfated subunit alphaV almost disappeared in the transfectants, but integrin subunit beta3 almost had no change. These results suggested that Gal3ST-2 was involved in tumor metastasis process by regulation of adhesion ability to selectins and expression of integrins.


Assuntos
Integrina alfaV/biossíntese , Integrinas/biossíntese , Neoplasias Laríngeas/metabolismo , Selectinas/biossíntese , Sulfotransferases/fisiologia , Western Blotting , Adesão Celular , Linhagem Celular Tumoral , Células Cultivadas , Regulação para Baixo , Endotélio Vascular/citologia , Endotélio Vascular/patologia , Humanos , Imunoprecipitação , Integrina alfaV/química , Integrina alfaVbeta3/química , Integrina alfaVbeta3/metabolismo , Integrinas/metabolismo , Selectina L/biossíntese , Neoplasias Laríngeas/patologia , Lectinas/metabolismo , Metástase Linfática , Microscopia de Fluorescência , Metástase Neoplásica , Plasmídeos/metabolismo , Interferência de RNA , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção
17.
Artigo em Inglês | MEDLINE | ID: mdl-12174308

RESUMO

We studied the activities of calcium-independent phosphatidylcholine-specific phospholipase C (PC-PLC) and the relationship between PC-PLC and gamma-GT in hepatoma cells. We noted that PC-PLC activity decreased significantly during hepatocarcinogenesis and the proliferation of CBRH-7919 cells, but increased significantly during differentiation. There was a close relationship between PC-PLC and gamma-GT activities. When CBRH-7919 cells were treated with exogenous PC-PLC, it took only 15 minutes for gamma-GT activity to decrease significantly and to the lowest level at 24 hours. However, gamma-GT activity in culture medium increased within 2 hours, then decreased afterward. We conclude that PC-PLC may regulate gamma-GT via certain pathways.

18.
Artigo em Inglês | MEDLINE | ID: mdl-12215789

RESUMO

The correlation between the dynamic changes of l,2-diacylglyceride (DAG), phospholipids and phospholipase during the diethylnitrosoamine-induced hepatocarcinogenesis in rat was studied. It was found that liver DAG increased to the first peak at week 8 and maintained on a higher than normal level until week 14 when the second highest peak appeared, then gradually dropped. The composition of phospholipid was estimated simultaneously in order to elucidate the origin of DAG. It was discovered that only phosphatidylcholine (PC) and phosphatidylinositides decreased during liver carcinogenesis, suggesting these two phospholipids may be the origin of the increased DAG. PC was probably the major source of DAG, owing to the decreased amount of PIs which was not sufficient to generate the increased amount of DAG. Then the activities of PC-specific phospholipase C (PC-PLC) and phospholipase D (PLD) were determined by means of enzyme-coupling colorimetric methods. The results showed that the marked elevation of these two enzymes at week 8 might be related to the first peak of. DAG concentration and the PC hydrolysed by PC-PLC might be responsible for the second peak of DAG at week 14. The decline of DAG after week 14 was possibly due to the gradually decrease of PLD activity beginning from week 10, but other unknown factors might also be responsible for this decline. The regulation of PC-PLC and PLO by protein kinase C and related oncogenes were discussed.

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