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1.
Chinese Journal of Pathology ; (12): 604-608, 2013.
Artigo em Chinês | WPRIM | ID: wpr-233383

RESUMO

<p><b>OBJECTIVE</b>To compare morphological differences of three drug-resistant hepatocellular carcinoma (HCC) cell subclones (Huh-7/ADM, Huh-7/CBP, Huh-7/MMC) and their parental Huh-7 cell line, to analyze differential microRNA (miRNA) expression profiles in these cells and, finally to screen for the abnormal expressed miRNAs in drug-resistant HCC cells.</p><p><b>METHODS</b>Cellular morphology was observed by histology and transmission electron microscopy. MiRNA microarray was used to analyze the differential miRNA expression profiles in these cells (Huh-7, Huh-7/ADM, Huh-7/CBP, Huh-7/MMC) followed by real time quantitative PCR validation.</p><p><b>RESULTS</b>The drug-resistant cells had more intracytoplasmic organelles and were larger in size along with increased cytological pleomorphism than the parental Huh-7 cells. Compared with the parental Huh-7 cells, 32 simultaneously up-regulated and 22 down-regulated miRNAs were found in three drug-resistant cells. Up-regulation of miR-15a, miR-16, miR-27b, miR-30b, miR-146a, miR-146b-5p, miR-181a, miR-181d and miR-194 was verified by RT-qPCR.</p><p><b>CONCLUSION</b>Drug-resistant HCC cells have abnormal expressed miRNAs, which may be explored to further investigate the association of miRNA expressions with multidrugs resistance in HCC.</p>


Assuntos
Humanos , Antineoplásicos , Farmacologia , Carboplatina , Farmacologia , Carcinoma Hepatocelular , Genética , Patologia , Linhagem Celular Tumoral , Doxorrubicina , Farmacologia , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Perfilação da Expressão Gênica , Neoplasias Hepáticas , Genética , Patologia , MicroRNAs , Genética , Metabolismo , Mitomicina , Farmacologia , Análise de Sequência com Séries de Oligonucleotídeos
2.
Chinese Journal of Hepatology ; (12): 925-929, 2012.
Artigo em Chinês | WPRIM | ID: wpr-246761

RESUMO

<p><b>OBJECTIVE</b>To investigate the impact of expression of kisspeptin-1 (KiSS-1) metastasis-suppressor gene on the proliferative, adhesive and invasive abilities of human hepatocellular carcinoma (HCC) using an in vitro cell system.</p><p><b>METHODS</b>The highly metastatic human hepatoma cell line MHCC97-H was transiently transfected with the pcDNA3.1/HisC vector expressing the KiSS-1 gene (experimental group) or the vector without the KisS-1 gene (blank control group). Untransfected cells served as the negative control group. Proliferative abilities of the three groups were assessed by flow cytometry and MTT assay. Adhesive abilities were assessed by MTT assays using matrigel and fibronectin. Invasive abilities and cell motility were assessed by chemoinvasion chamber assay using reconstituted matrigel and migration chamber assay using polycarbonate filters, respectively.</p><p><b>RESULTS</b>The experimental group showed significantly lower adhesion capacity to matrigel (0.257+/-0.029) than either the blank control group (0.374+/-0.016; t=-7.90345, P less than 0.01) or the negative control group (0.394+/-0.031; t=-7.22752, P less than 0.01). Similarly, the experimental group showed significantly lower adhesion capacity to fibronectin (0.292+/-0.004) than either the blank control group (0.394+/-0.010; t=-20.93138, P less than 0.01) or the negative control group (0.412+/-0.023; t=-11.31371, P less than 0.01). The experimental group also showed significantly lower numbers of cells with invasive capacity (42.40+/-1.14) than either the blank control group (66+/-1.58; t=-27.0711, P less than 0.01) or the negative control group (67.80 +/- 1.92; t=-25.4, P less than 0.01). Similarly, the experimental group showed significantly lower numbers of cells with chemotactic movement (65.80+/-1.92) than either the blank control group (93.80+/-2.28; t=-30.11750, P less than 0.01) or the negative control group (96.40+/-2.07; t=-24.19142, P less than 0.01). The experimental group showed slightly, but not significantly, lower cell proliferation (0.644+/-0.027) than either the blank control group (0.669+/-0.022; t=-1.60371, P?>?0.05) or the negative control group (0.678+/-0.027; t=-1.97828, P?>?0.05). In addition, there were no obvious differences between the three groups in the amounts of cells arrested in either the G1 phase or the S phase.</p><p><b>CONCLUSION</b>KiSS-1 overexpression suppresses the adhesion, invasion and motility, but not the proliferation, of hepatoma carcinoma cells in vitro. These findings imply that KiSS-1 might represent a promising new candidate for gene therapy against human hepatocellular carcinoma.</p>


Assuntos
Humanos , Apoptose , Carcinoma Hepatocelular , Patologia , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Kisspeptinas , Genética , Neoplasias Hepáticas , Patologia , Invasividade Neoplásica , Transfecção
3.
Chinese Journal of Hepatology ; (12): 356-361, 2011.
Artigo em Chinês | WPRIM | ID: wpr-290594

RESUMO

<p><b>OBJECTIVE</b>To study the expression of mPGES-1 in hepatocellular carcinoma (HCC), observe the effect of MK886 on down-regulation of mPGES-1 gene expression on the biology of human hepatocarcinoma cell line HepG2 and to investigate its significance in the occurrence, progression, metastasis and invasion.</p><p><b>METHODS</b>HCC tissues, para-carcinoma tissues, far-carcinoma tissues and normal liver tissues were collected. The expressions of mPGES-1 were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The proliferation, adherence, migration and invasion abilities of HepG2 cells interfered by MK886 were assessed by MTT and transwell technique respectively.</p><p><b>RESULTS</b>The expression of mPGES-1 in HCCs was higher than that in normal liver tissues (P < 0.01), which increased following histological grade. Furthermore, mPGES-1 expression level was higher in the capsule invasion and metastasis tumor than in primary locus. A significant dose-dependent down-regulation of expressions of mPGES-1 gene mRNA and protein were observed in HepG2 cells when MK886 was given for 48 h (F = 140.402, P < 0.01; a'= 0.00714, P < 0.01). Compared with the control group, the growth inhibitory rate of HepG2 cell was observed significantly time and dose-dependent when MK886 was given. The rate of adhesion cells in experimental groups were 85.3% ± 1.3%, 70.5% ± 1.5% and 45.8% ± 2.4%, respectively, less than that in control group 100.0% ± 0 (F = 626.313, P < 0.01). The migration cells was 92.47 ± 1.90, 62.63 ± 1.96 and 37.33 ± 0.83 respectively in the experimental groups after 24 h, lower than that in the control group 128.93 ± 2.60 (F = 1253.805, P < 0.01). The invasion assay revealed that the invading cells were 41.67 ± 1.30, 25.47 ± 1.30 and 13.93 ± 1.66 in the experimental groups, in contrast to 55.67 ± 2.08 in control group after 24 h. The difference between these groups was significant (F = 372.615, P < 0.01). The numbers of adhesion, migration and invasion of HepG2 cells were dose-dependent in MK886 groups.</p><p><b>CONCLUSION</b>Over-expression of mPGES-1 was associated with the tumorigenesis and progression of HCC. The down-regulation of mPGES-1 gene expression might indicated the decrease of the invasion and metastasis of HCC.</p>


Assuntos
Feminino , Humanos , Masculino , Carcinoma Hepatocelular , Metabolismo , Patologia , Adesão Celular , Movimento Celular , Proliferação de Células , Células Hep G2 , Indóis , Farmacologia , Oxirredutases Intramoleculares , Metabolismo , Neoplasias Hepáticas , Metabolismo , Patologia , Microssomos , Metabolismo , Invasividade Neoplásica , Metástase Neoplásica , Prostaglandina-E Sintases
4.
Chinese Journal of Pathology ; (12): 330-333, 2011.
Artigo em Chinês | WPRIM | ID: wpr-261789

RESUMO

<p><b>OBJECTIVE</b>To investigate the relationship between the expression of thymidine phosphorylase (TP) and the sensitivity of gastric carcinoma to 5-fluorouracil (5-FU) and its prodrugs.</p><p><b>METHODS</b>Gastric carcinoma cell line AGS was transfected with recombinant plasmid pEGFP-N1-TP or control plasmid pEGFP-N1 by lipofectamin 2000. The expression of green fluorescence labeled protein was observed under fluorescence microscope. Positive clones AGS-p and AGS-pTP were selected by G418 treatment. Expression of TP protein and mRNA was detected by immunocytochemistry and RT-PCR, respectively. Drug sensitivity to 5-FU and its prodrugs was assessed by MTT assay.</p><p><b>RESULTS</b>Cell clones with the expression of green fluorescent protein (AGS-p) and a clone with TP and green fluorescent fusion protein (AGS-pTP) were established. Immunostaining of TP protein was strongly positive in AGS-pTP and negative in AGS-p and AGS. The expression of TP mRNA was significantly higher in AGS-pTP (0.8090 ± 0.0450) than that in AGS (0.0490 ± 0.0046) and AGS-p (0.0610 ± 0.0069; P < 0.01). The sensitivity to doxifluridine and capecitabine in AGS-pTP was significantly increased, as compared with that in AGS-p. IC50 values of AGS-pTP to doxifluridine and capecitabine were estimated 1.7 folds and 2.2 folds as much as that of AGS-p, respectively. The sensitivity to 5-FU was not different between AGS-pTP and AGS-p.</p><p><b>CONCLUSIONS</b>Enhancement of TP expression improves the sensitivity of gastric carcinoma cells to doxifluridine and capecitabine. But it does not affect the sensitivity to 5-FU.</p>


Assuntos
Humanos , Antimetabólitos Antineoplásicos , Farmacologia , Capecitabina , Linhagem Celular Tumoral , Desoxicitidina , Farmacologia , Floxuridina , Farmacologia , Fluoruracila , Farmacologia , Plasmídeos , RNA Mensageiro , Metabolismo , Proteínas Recombinantes , Genética , Metabolismo , Sensibilidade e Especificidade , Neoplasias Gástricas , Metabolismo , Patologia , Timidina Fosforilase , Genética , Transfecção
5.
Chinese Journal of Hepatology ; (12): 357-360, 2010.
Artigo em Chinês | WPRIM | ID: wpr-326360

RESUMO

<p><b>OBJECTIVE</b>To study the relationship between FHIT and WWOX expression and clinicopathological features in hepatocellular carcinoma (HCC).</p><p><b>METHOD</b>The expression of FHIT and WWOX were determined by immunohistochemistry in 142 patients with HCC.</p><p><b>RESULTS</b>Absent or reduced FHIT and WWOX expression was observed in 68.3% and 77.5% of HCCs, respectively. The expression of FHIT was significantly correlated with that of WWOX (P < 0.01), and progressive loss of FHIT and WWOX expression were observed as tumor differentiation decreased and tumor grade increased (P < 0.05). Absent/reduced FHIT and WWOX expression was associated with tumor invasion and metastasis (P < 0.05). In addition, the expression of FHIT and WWOX in HCC with recrudesce were lower than that in those without recrudesce (P < 0.05).</p><p><b>CONCLUSIONS</b>Absent/reduced FHIT and WWOX expression is associated with poor prognosis.</p>


Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Hidrolases Anidrido Ácido , Genética , Carcinoma Hepatocelular , Genética , Patologia , Perfilação da Expressão Gênica , Neoplasias Hepáticas , Genética , Patologia , Proteínas de Neoplasias , Genética , Oxirredutases , Genética , Prognóstico , Proteínas Supressoras de Tumor , Genética , Oxidorredutase com Domínios WW
6.
Chinese Journal of Surgery ; (12): 185-188, 2010.
Artigo em Chinês | WPRIM | ID: wpr-254817

RESUMO

<p><b>OBJECTIVE</b>To evaluate the impact of preoperative three-dimensional visualization and virtual liver surgery planning on hepatic resection.</p><p><b>METHODS</b>All relevant structures (livers, portal vein, hepatic veins, and tumors) were extracted from multislice CT scans of 142 cases treated from May 2007 to May 2009. By the liver surgery planning system software Liv 1.0, reconstruction and image analysis of the relevant structures was performed and virtual resections of liver were carried out. Data were correlated to intraoperative findings.</p><p><b>RESULTS</b>(1) Three-dimensional visualization revealed the spatial relationship of tumors to the intrahepatic vascular system, thus giving impressions how the neoplasms were situated. Virtual tumor resections corresponded to the intraoperative findings. (2) With the planning, an intended resection could be performed virtually and optimal identification of resection margins could be achieved. The ischemia and congestion territory within the remaining liver parenchyma could be calculated. Simulation resections could avoid liver parenchyma over resection and maintain a sufficient amount of liver tissue to sustain hepatic function. Virtual simulations of tumor resection were used successfully to plan of surgical procedures in the hepatic tumors. Hepatectomy was performed in 29 cases after virtual tumor resections but seemed impossible with conventional CT scan. Resection plans of 92 cases were optimized after virtual resections. (3) The mean liver volume of patients with primary hepatocellular carcinoma measured by the software and the real resected was (477 +/- 223) ml and (451 +/- 209) ml respectively. Comparison by means of linear regression analysis between volume measurement on the software and the real resected showed a nearly ideal correlation coefficient (R = 0.922, P < 0.01). The mean error was 6.1%.</p><p><b>CONCLUSIONS</b>The three-dimensional tumor visualization and virtual simulation of tumor resections of the software Liv 1.0 provide an important reference for a valuable planning of complex hepatic resections. It is not only benefit to improve the predictability and security of hepatectomy but also helpful to improve the success rate of complex hepatic resections.</p>


Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Simulação por Computador , Hepatectomia , Métodos , Imageamento Tridimensional , Fígado , Diagnóstico por Imagem , Cirurgia Geral , Neoplasias Hepáticas , Cirurgia Geral , Tomografia Computadorizada por Raios X , Interface Usuário-Computador
7.
Chinese Journal of Hepatology ; (12): 17-20, 2008.
Artigo em Chinês | WPRIM | ID: wpr-277619

RESUMO

<p><b>OBJECTIVE</b>To detect the expression of survivin protein, survivin mRNA, p27 protein, p27 mRNA and PTEN protein in hepatocellular carcinomas (HCC) and their clinical significances.</p><p><b>METHODS</b>Tissue microarrays were constructed. The expression of survivin protein, p27 protein and PTEN protein were evaluated by immunohistochemical methods and in expression of survivin mRNA and p27 mRNA were evaluated by in stiu hybridization respectively in tumor tissues from 141 HCC patients, 128 samples of para-carcinoma liver tissues, 97 liver tissues far from the carcinomas and normal liver tissues from non HCC patients. The relationship of survivin, p27 and PTEN were investigated and a prediction model of HCC was constructed.</p><p><b>RESULTS</b>The expressions of survivin protein (Ridit 95% CI = 0.689+/-0.048, P < 0.01), survivin mRNA (Ridit 95% CI = 0.690+/-0.049, P < 0.01) and p27 protein (Ridit 95% CI = 0.556+/-0.053, P < 0.05) in HCC tissues were significantly increased, while the expression of PTEN protein (Ridit 95% CI = 0.282+/-0.048) in HCC tissues was significantly reduced (P < 0.01).</p><p><b>CONCLUSION</b>Overexpressions of survivin mRNA and p27 protein and reduced expression of PTEN protein might be a valuable marker to predict the presence of HCC.</p>


Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Hepatocelular , Metabolismo , Patologia , Inibidor de Quinase Dependente de Ciclina p27 , Proteínas Inibidoras de Apoptose , Peptídeos e Proteínas de Sinalização Intracelular , Metabolismo , Neoplasias Hepáticas , Metabolismo , Patologia , Proteínas Associadas aos Microtúbulos , Metabolismo , PTEN Fosfo-Hidrolase , Metabolismo , RNA Mensageiro , Genética
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