-
[show abstract]
[hide abstract]
ABSTRACT: To measure the expression pattern of STAT2 in cervical cancer initiation and progression in tissue sections from patients with cervicitis, dysplasia, and cervical cancer.
Antibody against human STAT2 was confirmed by plasmids transient transfection and Western blot. Immunohistochemistry was used to detect STAT2 expression in the cervical biopsies by using the confirmed antibody against STAT2 as the primary antibody.
It was found that the overall rate of positive STAT2 expression in the cervicitis, dysplasia and cervical cancer groups were 38.5%, 69.4% and 76.9%, respectively. The STAT2 levels are significantly increased in premalignant dysplasia and cervical cancer, as compared to cervicitis (P< 0.05). Noticeably, STAT2 signals were mainly found in the cytoplasm, implying that STAT2 was not biologically active.
These findings reveal an association between cervical cancer progression and augmented STAT2 expression. In conclusion, STAT2 increase appears to be an early detectable cellular event in cervical cancer development.
Asian Pacific Journal of Tropical Medicine 09/2012; 5(9):738-42. · 0.37 Impact Factor
-
Bo Li,
Xiang Li,
Yingxin Li,
Hui Guo,
Shu-Yan Sun,
Qiong-Qiong He,
Ying Wang,
Junming Luo,
Ji-Fang Wen,
Hui Zheng, De-Yun Feng
[show abstract]
[hide abstract]
ABSTRACT: Hepatitis C virus (HCV) infection has become a severe health problem worldwide. The viral proteins are believed to be among the most important factors that contribute to HCV mediated pathogenesis. Accumulated evidence demonstrating that HCV non-structural protein 3 (NS3) possesses oncogenic potential, and is involved in the regulation of cell proliferation has been documented. In this study, we emphasized the effect of HCV NS3 protein on cell proliferation in the immortally normal hepatocyte QSG7701 cells. The cell line transfected with plasmid expressing NS3 protein showed enhanced cell growth, extracellular signal-related kinase (ERK) activation, DNA binding activities of transcription factors of activator protein 1 (AP-1) and NF-kappaB, and cyclin D1 overexpression, but without activation of Jun amino-terminal kinase or p38. Pre-treatment of NS3 protein expressing cells with ERK inhibitor, PD98059, blocked the activation of AP-1 and NF-kappaB, and inhibited cyclin D1 expression and cell proliferation. The results suggest that NS3-mediated cell growth occurs through activation of ERK/AP-1 and NF-kappaB/cyclin D1 cascades.
International Journal of Molecular Medicine 08/2010; 26(2):273-9. · 1.98 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To examine whether 2'-5'oligoadenylate synthetase (OAS) gene promoter can be specifically activated by hepatitis C virus (HCV)-core protein.
Human embryo hepatic cell line L02 was transfected with pcDNA3.1-core plasmid and selected by G418. Expression of HCV-core was detected by reverse transcription polymerase chain reaction and Western blotting. The OAS promoter sequence was amplified from the genomic DNA and inserted into pGL3-basic vector. The resultant pGL3-OAS-Luci plasmid was transiently transfected into L02/core cells and luciferase activity was assayed.
L02/core cell line stably expressing HCV-core protein was established. The pGL3-OAS-Luci construct exhibited significant transcriptional activity in the L02/core cells but not in the L02 cells.
HCV-core protein activates the OAS gene promoter specifically and effectively. Utilization of OAS gene promoter would be an ideal strategy for developing HCV-specific gene therapy.
World Journal of Gastroenterology 08/2009; 15(25):3178-82. · 2.47 Impact Factor
-
Fa-Qing Tang,
Chao-Jun Duan,
Da-Mao Huang,
Wei-Wei Wang,
Chun-Lei Xie,
Jing-Jing Meng,
Lei Wang,
Hai-Ying Jiang, De-Yun Feng,
Shang-Hui Wu,
Huan-Hua Gu,
Mo-Yu Li,
Fu-Liang Deng,
Zhi-Jun Gong,
Hui Zhou,
Yong-Hao Xu,
Can Tan,
Xin Zhang,
Ya Cao
[show abstract]
[hide abstract]
ABSTRACT: N,N'-Dinitrosopiperazine (DNP) induces nasopharyngeal carcinoma (NPC) and shows organ specificity to the nasopharyngeal epithelium. To investigate its mechanism, the rat NPC model was induced using DNP. Rat NPC and normal nasopharyngeal cells were obtained from the NPC model using laser capture. The total proteins from these cell samples were separated with two-dimension polyacrylamide gel electrophoresis techniques, and highly expressed proteins (> five-fold) were analyzed using matrix-assisted laser desorption/ionization time of flight and bioinformatics. The results showed that HSP70 and mucin 5B expression increased not only in rat NPC but also in atypical hyperplasia nasopharyngeal tissues, a precancer stage of NPC. High-expression of heat shock protein 70 (HSP70) and mucin 5B was further supported by western blot analysis. The immunofluorescence and western-blotting studies further showed that DNP induced the expression of HSP70 and mucin 5B in a dosage-dependent manner in normal nasopharyngeal epithelia cells. Our data indicate that DNP triggers over-expression of HSP70 and mucin 5B, and is involved in nasopharyngeal tumorigenesis. HSP70 and mucin 5B may be important targets in nasopharyngeal tumorigenesis induced by DNP.
Cancer Science 01/2009; 100(2):216-24. · 3.33 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the role of AP-1 in the secretion of Type I collagen in TGF-beta1-stimulated human lung fibroblasts.
Human lung fibroblasts cell line (HLF-02) was cultured, and then stimulated with 10 microg/L TGF-beta1 at different time points. Curcumin was added into the culture medium to inhibit the AP-1 activity before incubating with TGF-beta1. AP-1 DNA binding activity was assayed by electrophoretic mobility shift assay (EMSA), and the expression of Type I collagen was detected by Western blot and RT-PCR.
TGF-beta1 could induce the transcription and secretion of Type I collagen in HLF-02 cells(P<0.05). TGF-beta1 could upregulate the AP-1 DNA binding activity ( P<0.05). Curcumin ( 5, 10, 15, and 20 micromol/L) could inhibit the AP-1 DNA binding activity in TGF-beta1-stimulated cells (the inhibition ratio was 17.1%, 17.6%, 24.2%, and 31.3%; P<0.05). Curcumin (5, 10, 15, and 20 micromol/L) could also inhibit the secretion of Type I collagen significantly (the inhibition ratio was 62.1%, 58.8%, 62.1%, and 59.6%; P<0.05).
AP-1 is responsible for the secrection of TGF-beta1-induced Type I collagen in human lung fibroblasts.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 10/2007; 32(5):776-81.
-
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology 08/2007; 15(7):540-1.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the proteome of hepatocyte transformation by hepatitis C virus (HCV) nonstructural protein 3 (NS3).
Human hepatocyte line QSG7701 stably expressing HCV NS3 C-terminal deleted protein was constructed, which was named pRcHCNS3/QSG. Two-dimensional electrophoresis (2-DE) was used to separate the total protein of pRcHCNS3/QSG and pRcCMV transfected cells (pRcCMV/QSG) respectively. Differentially expressed protein spots were identified by mass spectrometry. Western blot confirmed the differentially expressed proteins.
2-DE profiles with high resolution and reproducibility were obtained. The average spots of pRcHCNS3/QSG and pRcCMV/QSG were (1183+/-77) and (1095+/-82) respectively, and (920+/-60) spots were matched. Twenty-one differentially expressed protein spots were chosen randomly and 15 were identified by mass spectrometry. Some proteins such as Ras, P38 and HD53 which were involved in signal transduction were increased in pRcHCNS3/QSG cells. Western blot also showed strong expression of phosphorylated P44/42 and P38 in pRcHCNS3/QSG cells. Other differentially expressed proteins were related to cell cycle regulation, immunoreaction, tumor invasion and metastasis, and liver metabolizability.
HCV NS3 might be involved in cell malignant transformation through affecting protein expression and signal transduction such as MAPK cascade. Further study on the signal transductions and their relationship would not only be helpful to explore the mechanism of HCV related HCC, but also provide a new idea for the molecular treatment of HCC.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 07/2007; 32(3):387-95.
-
[show abstract]
[hide abstract]
ABSTRACT: To explore the cross-talk between extracellular signal-regulated kinase (ERK) and nuclear factor (NF-kappaB) signal transduction pathways in the hepatocytes expressing hepatitis C virus nonstructural protein 3 (HCV NS3).
A cell line QSG7701/NS3, which stably expressed HCV NS3 protein, was constructed by transfecting plasmid pcDNA3.1-NS3 into a human immortalized hepatocyte line QSG7701. Before and after QSG7701/NS3 cells were treated by MEK inhibitor PD98059, the phosphorylation level of ERK and the expression of cyclin D1 protein were detected by Western blot; the DNA binding activities of activator protein 1 (AP-1) and NF-kappaB were evaluated with electrophoretic mobility shift assay (EMSA). Cell cycles were measured by flow cytometry (FCM); the effects of PD98059 on the cell proliferation were determined by MTT assay.
HCV NS3 protein could up-regulate the phosphorylation of ERK and the expression level of cyclin D1 in QSG7701 cells. PD98059 could inhibit the cell proliferation mediated by HCV NS3 protein, down-regulate the activities of AP-1 and NF-kappaB, and suppress the expression of cyclin D1.
The inhibition of ERK can suppress the DNA binding activity of NF-kappaB and the cell proliferation mediated by HCV NS3 protein in a dose-dependent manner. There may be a cross-talk between ERK and NF-kappaB signal transduction pathways, which may exert synergistic action on the proliferation and malignant transformation of hepatocytes.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 05/2007; 32(2):259-63.
-
[show abstract]
[hide abstract]
ABSTRACT: To study the expression and localization of nuclear transcription factor Sp1 in macrophages after stimulated by silicon dioxide in vivo and in vitro.
Forty Sprague Dawley rats were randomly divided into the control group and the silica exposure group, 20 in each group. The rat silicosis models were established by direct tracheal instillation of silica into rat lung (0.2 g/kg) only once while the control group was instilled with equal amount of saline. Animals were killed at 1st, 7th, 14th, 21st and 28th day after instillation. Dynamic changes of Sp1 protein expression and its cellular localization were detected by immunohistochemistry in pulmonary macrophages. In vitro, Sp1 mRNA and protein expression and their dynamic changes were monitored by RT-PCR and western blotting after stimulated by silicon dioxide in cultured RAW264.7 macrophages respectively. Cellular localization of Sp1 protein was characterized by immunocytochemistry.
Compared to the control group, the Sp1 protein expression was increased in pulmonary macrophages and reached the peak at the 14th day in the silica exposure group. In vitro, the Sp1 mRNA level began to rise at 30 minutes after the administration of silicon dioxide and reached the peak at 240 minutes and then decreased to the minimal level at 960 minutes. The Sp1 total protein and nuclear protein also exhibited the similar trend. The former reached the peak at 240 minutes and the latter at 480 minutes. The significant nuclear translocation of Sp1 protein was observed at 120 minutes after the administration of silicon dioxide and became most significant at 480 minutes.
Silicon dioxide can activate nuclear transcription factor Sp1 in macrophages in vivo and in vitro. Sp1 might play an important pathogenic role in the development of silicosis.
Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases 10/2006; 24(9):518-22.
-
[show abstract]
[hide abstract]
ABSTRACT: To detect the effect of erigeron breviscapus on the expression of vascular endothelial growth factor (VEGF) in the periodontal tissues during orthodontic tooth movement.
45 rabbits were divided into 3 groups (groups A, B and C). Groups A and B included experimental group of 1, 3, 7 and 14 days respectively. The mandibular first molar of each experimental rabbit was observed. The rabbits of group A and group B received iontophoresis with erigeron breviscapus into the right (group A-R and group B-R) and with normal sodium into the left as the control (group A-L and group B-L). Additionally, the rabbits of group B were designed orthodontic appliance, by which 0.78 N mesial force was applied to pull the mandibular first molars. Group C, group of 0 day, was no iontophoresis and orthodontic appliance as the control. After killed on schedule, the amount of experimental tooth movement was measured and the expression of VEGF was examined by immunohistochemical method.
The amount of experimental tooth movement increased successively from 1 to 14 days. The differences among days 3, 7 and 14 were significant in the comparison between group B-R and group B -L (P < 0.01). The expression of VEGF in groups A-R and B-L enhanced apparently compared with that in groups C and A-L (P < 0.01), but that in group B-R was the most apparent (P < 0.01). The expression of VEGF reached the peak level on day 3 in groups A-R and B-R (P < 0.01), but it reached the peak level on day 7 in group B-L (P < 0.01).
Erigeron breviscapus by iontophoresis can accelerate orthodontic tooth movement, and can meanwhile up-regulate the expression of VEGF in periodontium in the earlier period of orthodontic tooth movement. Thus it can be presumed that one of its mechanisms for erigeron breviscapus to accelerate orthodontic tooth movement is erigeron breviscapus effects the metabolism and differentiation of osteoblast and osteoclast through up-regulating the expression of VEGF in periodontium.
Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology 10/2006; 24(5):458-61.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the relationship between the expression of Ang2, Tie2 and the angiogenesis of hepatocellular carcinoma in rats.
Thirty-eight healthy male rats were randomly divided into 3 groups: 5 rats in the control group; 25 rats in the experimental group were equally divided into 5-day, 10-day, 15-day, 20-day, and 25-day groups; the other 8 rats were used as the supplement of the experimental group. An allogenic transplanted rat model of CBRH-7919 hepatocellular carcinoma in situ was established by immunosuppression. The expressions of Ang2 and Tie2 were detected by immunohistochemical staining in cancerous tissues of different developmental stages and liver tissues of the control group. At the same time, microvessel density was determined by anti-CD31 immunohistochemical staining.
CBRH-7919 hepatocellular carcinoma models were successfully set up in 24 rats. The expression level of Ang2 and Tie2 in cancerous tissues was much higher than that of liver tissues of the control group (P <0.05). The overexpression of Ang2 was pristine and continuous in different developmental stages. The expressions of Ang2 and Tie2 positively correlated with microvessal density in hepatocellular carcinoma (P<0.05).
The up-regulation of Ang2 and Tie2 may play important roles in the angiogenesis of hepatocellular carcinoma. Ang2 may participate in the start of angiogenesis of hepatocellular carcinoma.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 08/2006; 31(4):523-7.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the role of TGF-beta(1)/MAPK signaling pathways in the expression of type I collagen and activity of MMP-2, 9 in human lung fibroblasts.
Human lung fibroblasts cell line (HLF-02) was cultured and and then stimulated with 10 ng/ml TGF-beta(1) for different time; SB203580 or PD98059 was added into culture medium to block p38 or ERK kinase pathway before incubated with TGF-beta(1); the expression of type I collagen was detected by Western blotting and RT-PCR; zymogram analysis was used to analyze the activity of MMP-2 and MMP-9.
(1) In the process of stimulation by TGF-beta(1), the type I collagen mRNA level of 24 h, 48 h and 72 h group was: 1.33 +/- 0.07, 2.46 +/- 0.09 and 2.39 +/- 0.08 respectively; and the type I collagen protein level of 24 h, 48 h and 72 h group was: 114.89 +/- 8.95, 208.16 +/- 6.75 and 211.46 +/- 8.05 respectively; and the activity of MMP-2 of 24 h, 48 h and 72 h group was: 190.33 +/- 5.86, 214.33 +/- 8.39 and 212.67 +/- 11.59 respectively. (2) SB203580 significantly inhibited the TGF-beta(1)-induced expression of type I collagen mRNA, protein and MMP-2 activity (inhibition ratio: 51%, 24% and 20%); (3) PD98059 also significantly attenuated the TGF-beta(1)-induced expression of type I collagen mRNA, protein and MMP-2 activity (inhibition ratio: 42%, 13% and 16%).
TGF-beta(1) is capable of inducing the expression of type I collagen mRNA and protein and up-regulating MMP-2 activity in HLF-02 cells. p38 and ERK kinase signaling pathways play important role in regulation and control for this process.
Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases 03/2006; 24(2):77-80.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the regulation of hepatitis C virus (HCV) core protein on the activity of signal transducers and activators of transcription 3 (stat3).
A cell line expressing stable HCV core protein-QSG7701-core was constructed by transfecting the pcDNA3. 1-core (expressing HCV core protein) into the human immortalized hepatocyte line QSG7701. The phosphorylation and DNA binding activity of stat3 were detected by immunocytochemistry, Western blot, and electrophoretic mobility shift assay (EMSA).
The expression level of phosphorylated stat3 in QSG7701-core cells was significantly lower than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells, but there were no significant differences in the expression levels of total stat3 among the 3 groups. The positive signal of phosphorylated stat3 in nucleus of QSG7701-core cells was obviously weaker than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells. EMSA showed that DNA binding activity of stat3 in QSG7701-core cells significantly decreased.
The expressionof HCV core protein in human hepatocyte line may suppress the phosphorylation and DNA binding activity of stat3, which may be one of the causes for resistance against interferon.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 01/2006; 30(6):631-5.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the relationship between infiltrating inflammatory cell and tumor angiogenesis in hepatocellular carcinoma (HCC) tissues and their clinicopathological features.
The paraffin-embedded specimens from 70 cases with HCC were stained using EliVision immunohistochemistry with mAbs against CD68, tryptase, and CD34. The counts of tumor-associated macrophage (TAM), mast cell (MC) and tumor microvessel (MV) were performed in the tissue sections.
The mean counts of TAM, MC, and MV in HCC tissues were significantly higher than those in pericarcinomatous liver tissues (TAM: 69.31+/-11.58 vs 40.23+/-10.36; MC:16.74+/-5.67 vs 7.59+/-4.18; MV: 70.11+/-12.45 vs 38.52+/-11.16, P<0.01). The MV count in the patients with metastasis was markedly higher than that with non-metastasis (P<0.01). In addition, the MC count in the patients with poorly differentiated HCC was obviously higher than that with well differentiated HCC (P<0.01). The correlation analysis showed that the TAM count was significantly correlated with the count of MV (r=0.712, P<0.01), and the MC count was obviously correlated with the MV count (r=0.336, P<0.05).
TAM and MC might be closely related to the enhancement of tumor angiogenesis. The MV count might be associated with tumor invasion and metastasis. Moreover, the MC count might be associated with tumor differentiation and prognosis of HCC.
World Journal of Gastroenterology 12/2005; 11(41):6521-4. · 2.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Survivin can block the cell apoptosis through inhibiting the functions of Caspase-3 and Caspase-7, and selectively overexpresses in common human cancers. This study was designed to investigate the effects of Survivin on tumorigenesis and development of oral cancer and its correlation to angiogenesis.
The expression of Survivin in 8 specimens of normal oral mucosa, 14 specimens of dysplastic leukoplakia and 47 specimens of oral squamous cell carcinoma (OSCC), and the expression of CD34 in the 47 specimens of OSCC was detected by SP immunohistochemistry. Microvessel density (MVD) was also assessed. The correlations of Survivin expression to MVD and clinicopathologic features of the OSCC patients were analyzed.
The positive rates of Survivin were 0% in normal oral mucosa, 14.24% in dysplastic leukoplakia, and 55.32% in OSCC. Survivin expression was significantly stronger in OSCC than in normal oral mucosa and dysplastic leukoplakia (P < 0.05), while there was no difference between the last 2 groups (P > 0.05). Survivin expression was significantly stronger in moderately and poorly differentiated OSCC than in well differentiated OSCC (P < 0.05); the positive rate of Survivin was significantly higher in OSCC with lymph node metastasis than in OSCC without lymph node metastasis(71.43% vs. 38.89%, P < 0.05). In OSCC, MVD was increased (25.87 +/- 12.10, 28.70 +/- 7.69, 35.42 +/- 10.09, 41.13 +/- 9.62, respectively) along with the strengthened Survivin expression (-, +, 2+, 3+) (P < 0.05).
Survivin expression is up-regulated in OSCC, and closely related with tumor cell differentiation, lymph node metastasis, and MVD; it may play an important role in tumorigenesis and development of OSCC.
Ai zheng = Aizheng = Chinese journal of cancer 11/2005; 24(11):1354-7.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the mechanism of silicosis by observing the effects of silica on the expression of plasminogen activator inhibitor-1 (PAI-1) and activator protein-1 (AP-1) in human alveolar epithelial cells type II (A549).
A549 cell and SiO(2) (200 microg /ml) were co-cultured for 0, 4, 8, 16 and 24 h respectively. The reverse transcriptase polymerase chain reaction (RT-PCR), Western blotting and SP immunocytochemistry were used for detections of the PAI-1 mRNA and protein expression. The nucleoprotein and total protein expression of AP-1 were investigated by Western blotting.
The expression levels of PAI-1 mRNA and protein were increased in a time-dependent manner(r(mRNA) = 0.911, r(protein) = 0.902, P < 0.05). The expressions of PAI-1 mRNA and protein in experimental groups were higher than that in control group (P < 0.05) and was the highest in 24 h group [(0.73 +/- 0.01) vs (0.36 +/- .03)]. The nucleoprotein expressions of c-jun/c-fos in experimental groups were also higher than in control group (P < 0.05), and the nucleoprotein expression level of c-jun was the highest in 4 h group [(1.54 +/- 0.02) vs (0.56 +/- 0.03)]; the nucleoprotein expression level of c-fos was the highest in 8 h group [(0.36 +/- 0.01) vs (0.15 +/- 0.01)]. Both c-jun and c-fos expression were decreased after 16 h, but the total protein expressionof c-jun/c-fos had no difference in all experimental groups. The positive signal of PAI-1 was located in cytoplasm and nucleus.
SiO(2) could induce PAI-1 expression of A549 in a time-dependent manner, and AP-1 activation can be observed in early time.
Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases 10/2005; 23(5):355-8.
-
[show abstract]
[hide abstract]
ABSTRACT: To explore the changes of apoptosis of cells in the lung tissue of rats with silica instillation and to its significance in silicosis, and to clarify the role of caspase-3 in the apoptosis progress.
Forty-eight rats were randomly divided into saline control groups and silica instillation groups, and the silicosis model was established in rats. Flow cytometry was used for detecting the rate of apoptosis at various stages. Immunohistochemistry for the expression of cleaved caspase-3.
The model of rat silicosis was established successfully. The apoptosis rate in the experimental group was significantly higher than that in the control group, and was increased with time. Caspase-3 was mainly expressed in alveolar epithelium cells, pulmonary macrophages and infiltrated inflammation cells. The expression of caspase-3 in the experimental group was stronger than that in the control group, but its expression intensity was not related to the cell apoptosis (r = 0.215, P > 0.05).
The apoptosis of the lung cells plays an important role during rat silicosis genesis. Caspase-3 plays an important role in regulating cell apoptosis during rat silicosis genesis.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 08/2005; 30(4):441-3.
-
[show abstract]
[hide abstract]
ABSTRACT: Cyclins overexpress in various tumors, but its expression in hepatocellular carcinoma (HCC) and its correlation to cell proliferation and apoptosis are unclear. This study was to determine the expression of Cyclins, proliferating cell nuclear antigen (PCNA), and cell apoptosis in HCC, and to analyze their interrelations.
Tissue microarray technology and SP immunohistochemistry were used to detect expressions of Cyclins A, B1, D1, E, and PCNA in 122 specimens of HCC. In situ terminal deoxyribonucleotide transferase labeling was used to detect cell apoptosis.
In the 122 HCC specimens, positive rate of Cyclin A was 50.0%, of Cyclin B1 was 47.5%, of Cyclin D1 was 42.6%, of Cyclin E was 35.2%. Cyclins levels were significantly higher in HCC tissues of grade II, III, and IV than in HCC tissues of grade I(P 0.05). Densities of apoptotic cells were significantly lower in HCC tissues of grade II, III, and IV than in HCC tissues of grade I(P 0.05); PCNA scores were significantly higher in HCC tissues of grade II, III, and IV than in HCC tissues of grade I(P 0.01). The poorer differentiation, the lower density of apoptotic cells in HCC, the higher PCNA score in HCC. Cyclins levels were negatively related to density of apoptotic cells (r=-0.686, P < 0.01), and positively related to PCNA score (r=0.599, P < 0.01); density of apoptotic cells was negatively related to PCNA score (r=-0.701, P < 0.01).
Cyclins overexpress in HCC, which may shorten tumor cell cycle phase, accelerate cell proliferation and decrease apoptosis, and result in increased malignant phenotypes.
Ai zheng = Aizheng = Chinese journal of cancer 07/2005; 24(6):695-8.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the effects of hepatitis C virus (HCV) core protein on the biological behaviors of human hepatocytes and their underlying mechanism.
A cell line expressing stably HCV core protein-QSG7701/core was constructed by transfecting the plasmid pcDNA3.1-core (expressing HCV core protein) into the human immortalized hepatocytes of the line QSG7701. The biological behaviors of these transfected cells were observed through plating-efficiency test, growth curve and flow cytometry (FCM). The association between HCV core protein and the expression of activated caspase-3 protein was evaluated by immunocytochemistry. The phosphorylation of mitogen-activate protein kinases (MAPKs) was detected with Western blotting. The activation of nuclear transcriptors AP-1, important effector molecule of MAPKs, and nuclear factor-kappa binding (NF-kappaB) were evaluated with luciferase assays and electrophoretic mobility shift assay (EMSA).
HCV core protein was expressed in the QSG7701/core cells and not in the QSG7701/pcDNA3.1 cells and untransfected QSG7701 cells. There were no significant differences in the expression levels of total P44/42(MAPK), p38(MAPK) and JNK among the QSG7701/core cells, QSG7701/pcDNA3.1 cells and untransfected QSG7701 cells. The expression levels of phophorylated P44/42(MAPK), p38(MAPK) and JNK in the QSG7701/core cells were significantly weaker than those in the QSG7701/pcDNA3.1 cells and untransfected QSG7701 cells. Plating efficiency test showed that the clone formation rate of the QSG7701/core cells was 32.25%, significantly lower than those of QSG7701/pcDNA3.1 and untransfected QSG7701 cells (47.5% and 42.5% respectively, both P < 0.01). The growth curve showed that the multiplication time of the QSG7701/core cells was 36 hours, significantly longer than those of the QSG7701/pcDNA3.1 and untransfected QSG7701 cells (27 and 28 hours respectively). FCM showed that the apoptotic rate of the QSG770/1core was 1.04%, lower than those of the QSG7701/pcDNA3.1 and untransfected QSG7701 cells (1.68% and 3.7% respectively), and that the percentage of theQSG770/1core cells at the G(0)/G(1) stage increased and those in the S stage decreased. Immunocytochemistry showed that the expression intensity of caspase-3 in the QSG7701/core cells was significantly weaker than those of the QSG7701/pcDNA3.1 cells and untransfected QSG7701 cells.
HCV core protein suppresses cell proliferation and apoptosis by downregmicrolating the phosphorylation of MAPKs and activating the transcriptors AP-1 and NF-kappaB, thus promoting the persistency of HCV infection which leads to chronic hepatitis C and hepatocellular cancer.
Zhonghua yi xue za zhi 05/2005; 85(18):1243-8.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the role of MAPK signal transduction in TGF-beta1 induced phenotypic differentiation of human lung fibroblasts.
Human lung fibroblasts cell line (HLF-02) were cultured and then stimulated with 10 ng/ml TGF-beta1 for different time; SB203580 or PD98059 was added into culture medium to prevent p38 or Erk kinase pathway before incubating with TGF-beta1; the expression of alpha-smooth muscle actin (alpha-SMA) was detected by Western blotting and RT-PCR; Western blotting was used to assay phosphorylation of p38, Erk, and JNK kinase.
(1) In the process of stimulation by TGF-beta1, the alpha-SMA mRNA expression levels of 24, 48 and 72 h groups were 1.87 +/- 0.11, 2.49 +/- 0.10, 3.02 +/- 0.15 respectively; and the alpha-SMA protein expression levels of 24, 48 and 72 h groups were 3.20 +/- 0.14, 3.96 +/- 0.21, 4.57 +/- 0.13 respectively. (2) TGF-beta1 induced p38, Erk kinase phosphorylation but not JNK kinase. (3) The inhibitors SB203580 and PD98059 suppressed TGF-beta1-induced p38 kinase and Erk phosphorylation respectively. (4) SB203580 significantly attenuated TGF-beta1-induced alpha-SMA mRNA and protein expression (inhibition rate: 30% and 40%); PD98059 also significantly inhibited TGF-beta1-induced alpha-SMA mRNA and protein expression (inhibition rate: 10% and 20%).
TGF-beta1 is capable of inducing the phenotypic differentiation of HLF-02, which is regulated by p38 and Erk kinase signal pathway.
Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases 05/2005; 23(2):109-12.
