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1.
Journal of Southern Medical University ; (12): 665-672, 2022.
Artigo em Chinês | WPRIM | ID: wpr-936361

RESUMO

OBJECTIVE@#To investigate the role of proline 4-hydroxylase Ⅱ (P4HA2) in the occurrence and progression of liver cancer.@*METHODS@#GEPIA and Human Protein Atlas database were used to predict the expression of P4HA2 in hepatocellular carcinoma (HCC), and K-M plotter online database was used to analyze the relationship between P4HA2 expression and the prognosis of HCC. We also examined the expressions of P4HA2 in HCC cells and normal hepatocytes using qRT-PCR and Western blotting. With lentivirus-mediated RNA interference, P4HA2 expression was knocked down in hepatoma SNU-449 and Hep-3B cells, and the changes in cell proliferation, migration and invasion were assessed using cell counting kit-8 (CCK-8) assay, colony formation test, scratch test and Transwell assay. The changes in the expressions of epithelial-mesenchymal transition (EMT) and PI3K/Akt/mTOR signal pathway-related proteins were detected using Western blotting.@*RESULTS@#Online database analysis showed that the expression of P4HA2 was significantly higher in HCC tissues than in normal liver tissues (P < 0.05). The expression levels of P4HA2 mRNA and protein were also significantly higher in HCC cell lines than in normal hepatocytes (P < 0.01). Lentivirus-mediated RNA interference of P4HA2 significantly lowered the expression levels of P4HA2 mRNA and protein in the hepatoma cells (P < 0.05) and caused obvious inhibition of cell proliferation, migration and invasion. P4HA2 knockdown significantly increased the expression of E-cadherin protein, lowered the expressions of N-cadherin and Snail, and obviously decreased the expressions of phosphorylated PI3K, AKT and mTOR (P < 0.05).@*CONCLUSION@#P4HA2 enhances the proliferation, migration, invasion, and EMT of hepatoma cells by activating the PI3K/Akt/mTOR signaling pathway to promote the occurrence and progression of liver cancer.


Assuntos
Humanos , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Hepáticas/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Prolil Hidroxilases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo
2.
Chinese journal of integrative medicine ; (12): 689-696, 2015.
Artigo em Inglês | WPRIM | ID: wpr-267168

RESUMO

<p><b>OBJECTIVES</b>To investigate the role of prolyl 4-hydroxylase beta polypeptide (P4HB) expressed in lung carcinoma and the intervention effect of Yiqi Chutan Formula (, YQCTF).</p><p><b>METHODS</b>Lung carcinoma model was established by subcutaneously inoculating LEWIS lung carcinoma cells in C57BL/6J mice. The differential expression of P4HB protein between the YQCTF (3.0 g/kg, gavage, once daily, 21 days) group and the control group was acquired by a 2 fluorescence difference gel electrophoresis (2D-DIGE), verified by Western blotting and identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS). The expression of P4HB and P4HB mRNA in cultured A549 cells from cisplatin (DDP) 1.5 µg/mL group and 15% serum combined with DDP 1.5 µg/mL group were detected by cellular immunohistochemistry and reverse transcription-polymerase chain reaction, respectively.</p><p><b>RESULTS</b>The proteomics research discovered that one-third of differential proteins including P4HB were decreased in the YQCTF group (P<0.01). Clinical pathology and tissue microarray studies showed that P4HB expression in lung cancer tissue was stronger than adjacent tissues and normal lung epithelial (P<0.01). In the YQCTF and DDP combined groups, the expression of P4HB and P4HB mRNA in A549 cell were decreased significantly (P<0.01).</p><p><b>CONCLUSION</b>YQCTF could inhibit the LEWIS lung carcinoma's growth, decrease the expression of P4HB in LEWIS lung carcinoma and A549 cells. YQCTF might take effect through regulating P4HB in endoplasmic reticulum to inhibit the incidence and growth process of lung carcinoma.</p>


Assuntos
Animais , Western Blotting , Carcinoma Pulmonar de Células não Pequenas , Tratamento Farmacológico , Genética , Patologia , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Medicamentos de Ervas Chinesas , Farmacologia , Usos Terapêuticos , Eletroforese em Gel Bidimensional , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , Neoplasias Pulmonares , Tratamento Farmacológico , Genética , Patologia , Camundongos Endogâmicos C57BL , Mapeamento de Peptídeos , Peptídeos , Farmacologia , Usos Terapêuticos , Prolil Hidroxilases , Genética , Metabolismo , Proteômica , RNA Mensageiro , Genética , Metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Análise Serial de Tecidos
3.
Korean Journal of Urology ; : 227-236, 2008.
Artigo em Coreano | WPRIM | ID: wpr-22623

RESUMO

PURPOSE: This study was performed to investigate prolyl 4-hydroxylase (P4H) expression changes and an inhibitor-mediated effect on bladder fibrosis in rats with partial bladder outlet obstruction(PBOO). MATERIALS AND METHODS: Twenty female Sprague-Dawley rats were divided into four groups. Group A(n=5) consisted of rats with PBOO treated with 2mg/kg P4H inhibitor, group B(n=5) consisted of rats with PBOO treated with 20mg/kg P4H inhibitor, group C(n=5) consisted of rats with PBOO treated with normal saline and group D(n=5) consisted of normal control animals. After PBOO for two weeks in the A, B, and C group rats, each amount of inhibitor was administered orally once a day for two weeks. After a total of four weeks, the bladders from all of the group rats were removed and evaluated. RESULTS: The muscle thickness calculated from Masson's trichrome staining was 0.85+/-0.22mm, 1.06+/-0.15mm, 1.19+/-0.30 and 0.49+/-0.10mm for group A, B, C, and D rats, respectively. The overall P4H expression was 65.7+/-15.2%, 13.4+/-8.4%, 73.8+/-15.5% and 10.0+/-10.0% for group A, B, C, and D rats, respectively. The overall collagen I protein expression was 16.9+/-18.0%, 17.0+/-24.1%, 30.5+/-13.4% and 8.8+/-8.7% for group A, B, C, and D rats, respectively. The overall collagen III protein expression was 9.6+/-4.2%, 8.8+/-2.9%, 12.5+/-10.6% and 7.5+/-3.5% for group A, B, C, and D rats, respectively. These results showed that PBOO led to increased muscle thickness and to an increased expression of P4H, collagen I and III protein, as compared with the group D control animals. Muscle thickness and expression of P4H, collagen I and III protein was decreased in rats treated with the P4H inhibitor-treated groups decreased as compared with rats in group C (saline-treated animals). The ratio of collagen I/III was 1.8, 1.9, 2.4 and 1.2 in group A, B, C, and D rats, respectively. CONCLUSIONS:Our results suggest that the P4H inhibitor may be potentially utilized to reduce bladder fibrosis caused by PBOO.


Assuntos
Feminino , Humanos , Ratos , Animais
4.
Journal of Veterinary Science ; : 105-109, 2006.
Artigo em Inglês | WPRIM | ID: wpr-32320

RESUMO

Recombinant human epidermal growth factor (rhEGF) stimulates the proliferation and migration of epithelial cells in human cell culture systems and animal models of partial-thickness skin wounds. This study investigated the effect of a topical rhEGF ointment on the rate of wound healing and skin re-epithelialization in a rat full thickness wound model, and verified whether or not the rhEGF treatment affected both myofibroblast proliferation and collagen synthesis in the dermis. When rhEGF (10 microgram/g ointment) was applied topically twice a day for 14 days, there was significantly enhanced wound closure from the 5th to the 12th day compared with the control (ointment base treatment) group. A histological examination at the postoperative 7th day revealed that the rhEGF treatment increased the number of proliferating nuclear antigen immunoreactive cells in the epidermis layer. In addition, the immunoreactive area of alpha-smooth muscle actin and the expression of prolyl 4-hydroxylase were significantly higher than those of the control group. Overall, a topical treatment of rhEGF ointment promotes wound healing by increasing the rate of epidermal proliferation and accelerating the level of wound contraction related to myofibroblast proliferation and collagen deposition.


Assuntos
Animais , Masculino , Ratos , Actinas/genética , Administração Tópica , Proliferação de Células/efeitos dos fármacos , Colágeno/biossíntese , Fator de Crescimento Epidérmico , Regulação da Expressão Gênica , Mioblastos Esqueléticos/efeitos dos fármacos , Antígeno Nuclear de Célula em Proliferação/genética , Ratos Sprague-Dawley , Cicatrização/efeitos dos fármacos
5.
Korean Journal of Pathology ; : 574-585, 1997.
Artigo em Coreano | WPRIM | ID: wpr-37743

RESUMO

This experiment was performed to elucidate the cytologic origin of chemically induced MFH in Wistar rats. The tumor was produced by injections of DMBA(9,10-dimethyl-1,2-benzanthracene). With the produced MFH, cell culture and cloning were performed, followed by establishment of a cell strain, which was investigated by immunohistochemical and electron microscopic studies. The results were as follows. A) By immunohistochemistry of the tumor tissue, fibroblastic cells were positive for MEP-1(specific antibody for fibroblastlike cell of MFH, Takeya, 1993) and Anti-hPH(beta)(Anti-prolyl 4-hydroxylase beta), but negative for TRPM-3 and F4/80. Histiocytelike cells were positive for TRPM-3 and F4/80, but negative for MEP-1 and Anti-hPH(beta). In immunoelectron microscopy, normal spleen macrophage showed linear reactivity in cell membrane for TRPM-3, whereas histiocytelike cells of the tumor disclosed negative reaction. B) At 5 weeks of the primary tumor cell culture, the cells exhibited typical storiform pattern of MFH. C) The established cell strain revealed immunoreactivity for MEP-1 and Anti-hPH(beta), but negative for TRPM-3. The cloned tumor cells showed morphologic characteristics of undifferentiated fibroblastic cell. Latex particle (0.80 micrometer size) phagocytosis was negative in the cloned cell strain. The results of the current study support the concept that principal component cells of MFH is of fibroblastic cell origin.


Assuntos
Animais , Ratos , 9,10-Dimetil-1,2-benzantraceno , Técnicas de Cultura de Células , Membrana Celular , Células Clonais , Clonagem de Organismos , Fibroblastos , Histiocitoma Fibroso Maligno , Imuno-Histoquímica , Macrófagos , Microscopia Imunoeletrônica , Microesferas , Fagocitose , Ratos Wistar , Baço
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