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1.
J Clin Lab Anal ; 32(6): e22430, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29516551

RESUMO

BACKGROUND: Hepatocellular carcinoma (HCC) is a malignant tumor that severely threatens human health. To date, early detection for HCC patients is particularly significant due to their poor survival rates even after liver resection. METHODS: Therefore, an efficient and sensitive detection method for monitoring liver cancer, multiplex methylation-specific PCR (MSP) coupled with capillary electrophoresis, is developed. RESULTS: Simulations demonstrated that the methylation status of RASSF1A, p16, SFRP1, and ELF could be detected even when DNA equaled or exceeded 12.5 ng simultaneously. Also, its accuracy for methylation detection outweighed polyacrylamide gel electrophoresis (87.5%) and agarose electrophoresis (84.3%), reaching 92.1%. Subsequently, we implemented multiplex MSP with capillary electrophoresis to investigate methylation status of the four tumor suppressor genes in tissue specimens and explore the prognostic value for HCC patients. As the data suggested, multivariate cox regression analysis revealed that the recurrence-free survival of 46 patients was greatly associated with portal vein tumor thrombus (PVTT) and p16 methylation and receiver operating characteristic (ROC) curves demonstrated that the predictive range of portal vein tumor thrombus (PVTT) combined with p16 hypermethylation was more sensitive than that of either PVTT or p16 hypermethylation alone with regard to disease recurrence in patients with HCC, which could be testified as a valuable biomarker in Clinical application. CONCLUSION: Multiplex MSP coupled with capillary electrophoresis has an excellent prospect of clinical application for monitoring early liver cancer and screening valuable biomarkers for prognosis of HCC patients.

2.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 49(1): 1-7, 2018 Jan.
Artigo em Zh | MEDLINE | ID: mdl-29737080

RESUMO

OBJECTIVE: To study the regulation of suppressor of cytokine signaling 3 (SOCS3) expression bythe brother of the regulator of the imprinted site (BORIS) in hepatocellular carcinoma cell. METHODS: The expression of SOCS3 mRNA in HCC cell lines was detected by real-time quantitative PCR (qRT-PCR). The expression of SOCS3 protein in knockdown and overexpression BORIS of HCC cell lines was tested by Western blot. The SOCS3 gene promoter methylation statusin the knockdown and overexpression BORIS of hepatocarcinoma cell lines was detected by using methylation specific PCR (MSP-PCR) method.The potential BORIS binding site of SOCS3promoter region was found by UCSC database analysis.The enrichment of BORIS in SOCS3 promoter region in endogenous high expression BORIS of HCC cells was evaluated by using chromatin immunoprecipitation (ChIP)-qPCR (ChIP-qPCR).The SOCS3 promoter region histone methylation status in the knockdown and overexpression BORIS of HCC was detected by ChIP-qPCR. RESULTS: The expression of SOCS3 mRNA in hepatocellular carcinoma cells was higher and SOCS3 protein expression was down-regulated or up-regulated in the knockdown or overexpression of BORIS mRNA hepatocarcinoma cells,so BORIS has a positive regulatory effect on SOCS3 protein expression in hepatocarcinoma cells. MSP-PCR experiments showed that the SOCS3 promoter in SMMC-7721 and HepG2 cells was unmethylated and knockdown of BORIS did not change the methylation status; the SOCS3 promoter region of Huh7 cells was methylated; after overexpression of BORIS,the SOCS3 promoter region was changed to an unmethylated state; the SOCS3 promoter was unmethylated in HCCLM3,overexpression of BORIS did not alter the methylation status. The ChIP-qPCR assay demonstrated that BORIS specifically binds to the SOCS3 promoter region in HCC cells with high expression of BORIS. Histone methylation assay indicated that knockdown of BORIS reduced BORIS enrichment in the SOCS3 promoter region, with decreasing H3K4 me2 and increasing H3K27 me3 in the region of histone,whereas the overexpress BORIS in HCC cells showed the opposite situation. CONCLUSION: BORIS plays a role of epigenetic regulationon SOCS3 gene promoter methylation and histone methylation,modulating the expression of SOCS3,and then involved in the development of hepatocellular carcinoma.


Assuntos
Carcinoma Hepatocelular/genética , Proteínas de Ligação a DNA/metabolismo , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/genética , Proteína 3 Supressora da Sinalização de Citocinas/metabolismo , Linhagem Celular Tumoral , Imunoprecipitação da Cromatina , Metilação de DNA , Histonas/metabolismo , Humanos , Regiões Promotoras Genéticas
3.
Plasmid ; 90: 15-19, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28189631

RESUMO

Lentiviral vectors are now widely considered as one of the most common gene delivery tools for dividing and non-dividing cells. However, insertional mutagenesis has been found in clinical trials with retroviral vectors, which poses a safety risk. The use of non-integrating lentiviral (NIL) vectors, which avoid integration, eliminates the insertional mutagenesis problem. These NIL vectors are unable to mediate stable gene delivery into dividing cells, which makes them of limited use in the clinical practice of gene therapy. In this study, we constructed a NIL vector which harbors the scaffold/matrix attachment region (S/MAR) sequence and a therapeutic gene. NIL retained episomal erythropoietin (EPO) gene expression for 74days in dividing cells both with and without selection. Furthermore, Southern blot analysis showed that the NIL vector was retained extrachromosomally in CHO cells. In conclusion, the NIL vector based on an S/MAR sequence retained the extrachromosomal expression of a therapeutic gene in dividing cells. Our results show that NIL vectors maybe a safe and effective means of gene delivery, which is of potential clinical significance.


Assuntos
Eritropoetina/genética , Técnicas de Transferência de Genes , Vetores Genéticos/metabolismo , Lentivirus/genética , Plasmídeos/metabolismo , Animais , Células CHO , Divisão Celular , Cricetulus , Eritropoetina/metabolismo , Expressão Gênica , Vetores Genéticos/química , Células HEK293 , Humanos , Lentivirus/metabolismo , Fator 1 de Elongação de Peptídeos/genética , Fator 1 de Elongação de Peptídeos/metabolismo , Plasmídeos/química , Regiões Promotoras Genéticas , Transgenes
4.
Mol Cancer ; 15(1): 40, 2016 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-27189061

RESUMO

BACKGROUND: Recent studies have suggested that cancer cells contain subpopulations that can initiate tumor growth, self-renew, and maintain tumor cell growth. However, for esophageal cancer cells, the relationship between STAT3, microRNAs and cancer stem cells remains unclear. METHODS: Serum-free culture was used to enrich esophageal cancer stem-like cells (ECSLC). Flow cytometry determined the proportion of ECSLC. qPCR were performed to examine expression level of stemness factors, mesenchymal markers, ATP-binding cassette (ABC) transporters, STAT3, miR-181b, CYLD. Western blot were performed to analyze the expression of STAT3, p-STAT3 and CYLD (cylindromatosis). BALB/c mice xenograft studies were conducted to evaluate the tumorigenicity of enriched ECSLC. Sphere formation assay and colony formation assays were employed to analyze the relationship between STAT3 and miR-181b. Luciferase assays were used to evaluate activity which CYLD is a target of miR-181b. RESULTS: Sphere formation cells (SFCs) with properties of ECSLC were enriched. Enriched SFCs in serum-free suspension culture exhibited cancer stem-like cell properties and increased single-positive CD44 + CD24-, stemness factor, mesenchymal marker expression ABC transporters and tumorigenicity in vivo compared with the parental cells. Additionally, we found that reciprocal activation between STAT3 and miR-181b regulated SFCs proliferation. Moreover, STAT3 directly activated miR-181b transcription in SFCs and miR-181b then potentiated p-STAT3 activity. Luciferase assays indicated that CYLD was a direct and functional target of miR-181b. CONCLUSION: The mutual regulation between STAT3 and miR-181b in SFCs was required for proliferation and apoptosis resistance. STAT3 and miR-181b control each other's expression in a positive feedback loop that regulates SFCs via CYLD pathway. These findings maybe is helpful for targeting ECSLC and providing approach for esophageal cancer treatments.


Assuntos
Neoplasias Esofágicas/genética , Neoplasias Esofágicas/metabolismo , MicroRNAs/genética , Células-Tronco Neoplásicas/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Regiões 3' não Traduzidas , Animais , Apoptose/genética , Sítios de Ligação , Linhagem Celular Tumoral , Proliferação de Células , Enzima Desubiquitinante CYLD , Modelos Animais de Doenças , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Interferência de RNA , Esferoides Celulares , Células Tumorais Cultivadas , Ensaio Tumoral de Célula-Tronco , Ensaios Antitumorais Modelo de Xenoenxerto
5.
Mol Cancer ; 14: 56, 2015 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-25890196

RESUMO

BACKGROUND: Acute myeloid leukemia (AML) is initiated and maintained by a subset of self-renewing leukemia stem cells (LSCs), which contribute to the progression, recurrence and therapeutic resistance of leukemia. However, the mechanisms underlying the maintenance of LSCs drug resistance have not been fully defined. In this study, we attempted to elucidate the mechanisms of LSCs drug resistance. METHODS: We performed reverse phase protein arrays to analyze the expression of anti-apoptotic proteins in the LSC-enriched leukemia cell line KG-1a. Immuno-blotting, cell viability and clinical AML samples were evaluated to verify the micro-assay results. The characteristics and transcriptional regulation of survivin were analyzed with the relative luciferase reporter assay, mutant constructs, chromatin immuno-precipitation (ChIP), quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR), and western blotting. The levels of Sp1, c-Myc, phospho-extracellular signal-regulated kinase (p-ERK), phospho-mitogen and stress-activated protein kinase (p-MSK) were investigated in paired CD34+ and CD34- AML patient samples. RESULTS: Survivin was highly over-expressed in CD34 + CD38- KG-1a cells and paired CD34+ AML patients compared with their differentiated counterparts. Functionally, survivin contributes to the drug resistance of LSCs, and Sp1 and c-Myc concurrently regulate levels of survivin transcription. Clinically, Sp1 and c-Myc were significantly up-regulated and positively correlated with survivin in CD34+ AML patients. Moreover, Sp1 and c-Myc were further activated by the ERK/MSK mitogen-activated protein kinase (MAPK) signaling pathway, modulating survivin levels. CONCLUSION: Our findings demonstrated that ERK/MSK/Sp1/c-Myc axis functioned as a critical regulator of survivin expression in LSCs, offering a potential new therapeutic strategy for LSCs therapy.


Assuntos
Resistencia a Medicamentos Antineoplásicos/genética , Proteínas Inibidoras de Apoptose/genética , Leucemia Mieloide Aguda/genética , Sistema de Sinalização das MAP Quinases/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Proteínas Proto-Oncogênicas c-myc/genética , Fator de Transcrição Sp1/genética , Adulto , Antígenos CD34/genética , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Feminino , Células HL-60 , Humanos , Células K562 , Masculino , Células-Tronco Neoplásicas/metabolismo , Transdução de Sinais/genética , Survivina , Transcrição Gênica/genética , Células U937 , Regulação para Cima/genética
6.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 46(5): 661-6, 2015 Sep.
Artigo em Zh | MEDLINE | ID: mdl-26619531

RESUMO

OBJECTIVE: To investigate the mechanisms of interaction between high-density lipoprotein binding protein (HDLBP)-VIGILIN with other proteins, we cloned VIGILIN cDNA N, KH1-7, KH8-12, KH13-14, and C fragments separately into expression vector, and identify the expressed proteins. METHODS: The recombinant plasmid pDsred2-N1/VIGILIN was used as template to amplify VIGILIN full length, VIGILIN N terminal, KH1- 7, KH8-12, KH13-14, C terminal and recombinated them with pGEX 5X 3. After transformed into E. coli BL21 cells, the recombinants were confirmed by enzyme digestion and sequence analysis. After optimizing the IPTG inducing condition, we induced GST-VIGILIN fusion proteins on the appropriate conditions. RESULT: The recombinant plasmids of pGEX 5X 3/VIGILIN FL, pGEX 5X 3/VIGILIN N terminal, pGEX 5X 3/VIGILIN KH1-7, pGEX 5X 3/VIGILIN KH8-12, pGEX 5X 3/VIGILIN KH13-14, pGEX 5X 3/VIGILIN C terminal were constructed successfully, and induced the GST-VIGILIN fusion proteins. CONCLUSION: pGEX 5X 3/VIGILIN FL, pGEX 5X 3/VIGILIN N terminal, pGEX 5X 3/VIGILIN KH1-7, pGEX SX 3/VIGILIN KH8-12, pGEX 5X 3/ VIGILIN KH13-14, pGEX 5X 3/VIGILIN C terminal recombinant plasmids were constructed successfully, and their corresponding fusion proteins were successfully expressed.


Assuntos
Proteínas de Ligação a RNA/genética , Clonagem Molecular , DNA Complementar/genética , Escherichia coli , Humanos , Lipoproteínas HDL/metabolismo , Plasmídeos , Proteínas de Ligação a RNA/biossíntese , Proteínas Recombinantes de Fusão/biossíntese
7.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 45(2): 196-201, 2014 Mar.
Artigo em Zh | MEDLINE | ID: mdl-24749339

RESUMO

OBJECTIVE: To explore the effects of CCCTC-binding factor (CTCF) on human liver cancer stem cells (HepG2) and cell proliferation of HepG2 and Nasopharyngeal carcinoma cell line (CNE1). METHODS: The pEGFP-N1/CTCF, CTCF-shRNA and GFP-shRNA plasmids were constructed and transfected into HepG2 and CNE1 cells, and RT-PCR or Western blot were performed to detect the mRNA or protein levels of CTCF. The subpopulation of CD90+ cancer stem cells in HepG2 cells transfected with CTCF-shRNA plasmid or GFP-shRNA plasmid (as transfection control) were assayed by flow cytometry with the wild type HepG2 cells as control. Proliferation of cells transfected with CTCF-overexpression or CTCF-shRNA plasmid was evaluated by MTT assay. RESULTS: The levels of both mRNA and protein of CTCF were increased in pEGFP-N1/CTCF transfected HepG2 and CNE1 cells compared to that in pEGFP-N1 transfected cells (P < 0.05), and decreased in CTCF-shRNA transfected cells compared to that in cells transfected with GFP-shRNA (P < 0.05). The results of flow cytometry demonstrated that, detection rate of CD90+ cells in cells transfected with CTCF-shRNA plasmid [(1.7330 +/- 0.4177)%] was obviously higher than that of wild-type HepG2 cells [(0.5750 +/- 0.0629)%] and cells transfected with GFP-shRNA plasmid [(0.3500 +/- 0.0866)%] (P < 0.05). The results of MTT analysis showed that, alteration of CTCF had no effect on cancer cell proliferation (P > 0.05). CONCLUSION: CTCF inhibits human liver cancer stem cells but no effect on cell proliferation.


Assuntos
Carcinoma Hepatocelular/metabolismo , Proliferação de Células , Neoplasias Hepáticas/metabolismo , Células-Tronco Neoplásicas/citologia , Proteínas Repressoras/metabolismo , Fator de Ligação a CCCTC , Linhagem Celular Tumoral , Citometria de Fluxo , Células Hep G2 , Humanos , Plasmídeos , RNA Mensageiro , RNA Interferente Pequeno , Transfecção
8.
Zhonghua Fu Chan Ke Za Zhi ; 48(8): 579-83, 2013 Aug.
Artigo em Zh | MEDLINE | ID: mdl-24199922

RESUMO

OBJECTIVE: To investigate the feasibility and effectiveness of human umbilical cord mesenchymal stem cells (HUCMSC) transplantation in the treatment of female stress urinary incontinence (SUI) in rats. METHODS: The 14 female SD rats of SUI model were established by vaginal balloon dilation after birth and maintain this status for four hours bilateral ovariectomy were performed after two weeks and were routinely reared for two months, then 12 SUI rat model were made. Two months later, transfected with plasmid pEGFP-N1 of HUCMSC were injected into the region surrounding the urinary tract matched with saline injection as control group. To get genitourinary tissue after testing urodynamic indicators, and observe the pathological changes of the bladder, urethra and the surrounding tissue; fluorescent cell of the experimental groups specimens were observed by fluorescence microscope. RESULTS: The leak point pressure(LPP) was (23.8 ± 4.2) mm Hg(1 mm Hg = 0.133 kPa) of the SUI rats. Transplanting mesenchymal stem cells of SUI rats, the positive rate of sneeze test was 1/6 in SUI group and 5/6 in control group, which reached statistical significance (P < 0.05); LPP was (30.6 ± 2.8) mm Hg in SUI group and (21.4 ± 7.0) mm Hg in control group, which reached statistical significance (P < 0.05) .In SUI rate model, connective tissue content were increased in urethra and the surrounding tissue and more fluorescent cell were observed. CONCLUSIONS: A rat model of female SUI was established successfully through postpartum vaginal balloon dilation and bilateral ovariectomy. MSC can be survived and proliferated in the urethral and the surrounding tissue of injured rats, and improve the urodynamic indicators and the positive rate of sneeze test. Morphology shows renovation of the support structures around the urethra.


Assuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Transplante de Células-Tronco Mesenquimais/métodos , Incontinência Urinária por Estresse/terapia , Animais , Proliferação de Células , Células Cultivadas , Modelos Animais de Doenças , Feminino , Humanos , Ovariectomia , Ratos , Ratos Sprague-Dawley , Transplante Heterólogo , Cordão Umbilical/citologia , Uretra/patologia , Uretra/fisiopatologia , Incontinência Urinária por Estresse/etiologia , Incontinência Urinária por Estresse/fisiopatologia , Urodinâmica , Vagina/lesões , Vagina/fisiopatologia
9.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 29(2): 176-80, 2012 Apr.
Artigo em Zh | MEDLINE | ID: mdl-22487828

RESUMO

CTCF as a multivalent eukaryotic transcription factor plays a diverse range of roles in regulation of various genes through the binding of its 11 zinc fingers to CTCF consensus sites or various proteins. CTCF is involved in multiple aspects of epigenetic regulation including regulation of chromatin remodeling and genomic imprinting. Deregulation of these processes result in a group of diseases are characterized by growth, development, and neurological dysfunction. This paper reviews recent researches that highlight the links between CTCF, epigenetics and diseases.


Assuntos
Proteínas Repressoras/genética , Fatores de Transcrição/genética , Fator de Ligação a CCCTC , Doença/genética , Epigenômica/métodos , Humanos , Dedos de Zinco/genética
10.
Exp Ther Med ; 23(3): 232, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35222709

RESUMO

Sorafenib is currently used to treat hepatocellular carcinoma (HCC). However, the development of chemoresistance to sorafenib is a major limitation for sorafenib-based therapy in patients with HCC. In the present study, the effect of the combination therapy of sorafenib and wh-4 on the proliferation of liver cancer cells was investigated. The results showed that sorafenib with wh-4 additively suppressed the proliferation of liver cancer cells. The colony formation of liver cancer cells decreased significantly in response to the combination treatment of sorafenib with wh-4, and it also induced the apoptosis of liver cancer cells. Western blot analysis demonstrated decreased expression of Bcl2, and increased expression of Bax in liver cancer cells treated with a combination of sorafenib and wh-4. Moreover, the migration of liver cancer cells was inhibited. The combination treatment of sorafenib with wh-4 reduced the expression levels of ABCB1 and ABCG2 which are responsible for resistance. Finally, STAT3 overexpression abolished the proliferation inhibition effect of sorafenib with wh-4 on liver cancer cells, and sorafenib and wh-4 suppressed the proliferation of liver cancer cells by STAT3 pathway. Together, these results suggest that sorafenib-wh4 combination treatment is a potential novel therapeutic approach to suppress the proliferation of liver cancer cells.

11.
Biosci Biotechnol Biochem ; 75(8): 1540-5, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21821931

RESUMO

SNX-2112 is a heat shock protein 90 (Hsp90) inhibitor with anticancer properties currently in clinical trials. This study investigated the effects of SNX-2112 on inhibition of cell growth, the cell cycle, and apoptosis in MCF-7 human breast cancer cells, in addition to the various molecular mechanisms. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometric analysis suggest that SNX-2112 inhibits cell growth in a time- and dose-dependent manner more potently than 17-(allylamino)-17-demethoxygeldanmycin (17-AAG), a traditional Hsp90 inhibitor, probably as a result of cell-cycle arrest at the G2/M phase and the induction of apoptosis. Downregulation of Bcl-2 and Bcl-xL, upregulation of Bax, cleavage of caspase-9 and poly (ADP-ribose) polymerase (PARP), and degradation of the breast cancer-related Hsp90 client proteins human epidermal growth factor receptor-2 (HER2), Akt, Raf-1, and nuclear factor kappa-B kinase (IKK) were observed in SNX-2112 treated cells by Western blot assay. These findings suggest that the molecular mechanisms of cell-growth inhibition by SNX-2112 involve activation of the mitochondrial apoptotic pathway and the degradation of breast cancer-related proteins.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico HSP90/antagonistas & inibidores , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Mitocôndrias/efeitos dos fármacos , Benzoquinonas/farmacologia , Western Blotting , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Feminino , Proteínas de Choque Térmico HSP90/metabolismo , Humanos , Lactamas Macrocíclicas/farmacologia , Mitocôndrias/genética , Mitocôndrias/metabolismo , Poli(ADP-Ribose) Polimerases/genética , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-raf/genética , Proteínas Proto-Oncogênicas c-raf/metabolismo , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , Proteína bcl-X/genética , Proteína bcl-X/metabolismo
12.
Artigo em Zh | MEDLINE | ID: mdl-21137318

RESUMO

2009 marks the 100th anniversary of the first description of Pneumocystis, an organism that was initially classified as a protozoan and later in 1988, was phylogenetically reclassified in the fungal kingdom. The organism was ignored for its first 50 years but that has subsequently been recognized as an important pathogen in immunocompromised patients, especially patients with acquired immune deficiency syndrome(AIDS). This paper reviewed recent progress on the molecular biology, involving in the life cycle and metabolism of the organism.


Assuntos
Pneumocystis/genética , Biologia Molecular , Pneumocystis/metabolismo
13.
Front Pharmacol ; 11: 532395, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33390934

RESUMO

Many studies have demonstrated that cancer stem cells (CSCs) or tumor-initiating cells (TICs) are responsible for tumor cell proliferation, chemotherapy resistance, metastasis, and relapse in various cancers. We, and others, have previously shown that the signal transducer and activator of transcription 3 (STAT3) signaling pathway is responsible for CSCs and TICs growth. Recent reports have indicated that the heat shock protein 90 (Hsp90) is also essential for the survival of CSCs and TICs. SNX-2112 is an Hsp90 inhibitor. However, it remains unclear whether proliferation of esophageal cancer stem-like cells (ECSLCs) is suppressed by SNX-2112 with knockdown of STAT3 (shSTAT3). Here, we explored the association between SNX-2112 with shSTAT3 and the suppression of ECSLCs growth. We found that the expression level of both STAT3 and p-STAT3 was higher in clinical esophageal cancer tissue than in the adjacent normal tissue, using western blot and qPCR analysis. Furthermore, differential expression analysis demonstrated that STAT3 was overexpressed in clinical specimens. We demonstrated that SNX-2112 inhibited cancer cell proliferation, decreased ABCB1 and ABCG2 gene expression levels and reduced the colony formation capacity of ECSLCs, which was enhanced by STAT3 silencing. Flow cytometry analysis revealed that the combination of SNX-2112 and shSTAT3 significantly induced apoptosis and cell cycle arrest at G2/M phase in ECSLCs. Levels of proliferation pathway proteins, including p38, c-Jun N-terminal kinase (JNK), and extracellular signal-regulated kinase (ERK) which were also client proteins of Hsp90, were also reduced. In addition, SNX-2112 with shSTAT3 inhibited the proliferation of ECSLCs in vivo. Finally, STAT3 overexpression eliminated the apoptotic and antiproliferative effects of SNX-2112 on ECSLCs. Hence, these results provide a rationale for the therapeutic potential of the combination of SNX-2112 with shSTAT3 in esophageal cancer, and may indicate new targets for clinical intervention in human cancer.

14.
Zhong Yao Cai ; 32(3): 384-8, 2009 Mar.
Artigo em Zh | MEDLINE | ID: mdl-19565717

RESUMO

OBJECTIVE: To study the effect of Berberine on proliferation, differentiation and apoptosis of K562 cells for supplying the theoretical evidence on the clinical application of Coptis chinensis to cure chronic myeloid leukemia. METHODS: The proliferation-inhibiting capability of K562 cells was investigated by MTT assay and colony conform test. The apoptosis effect of Berberine on K562 cells were analyzed by Wright's-Giemsa staining; DNA fragmentation was performed by agarose gel electrophoresis. The cell cycle distribution and the cell surface marker-cluster of differentiation were determined by flow cytometry. RESULTS: Berberine effectively inhibited the proliferation of K562 cells in a dose and time-dependent manner. In addition, when combined with different concentrations of Ara-C, the cells' viability percentage was prominently higher than only Berberine used. Treated with Berberine for 48 hours, the cells could be induced differentiation towards erythrocyte, granulocyte and megakaryocyte and with the treated time extending, the percentage of apoptosis cells gradually increased. CONCLUSIONS: Berberine can efficiently inhibit the proliferation of K562 cells, maybe by the way of blocking cells at the stage of G0/G1 and (or) G2, then leading to its apoptosis and differentiation.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Berberina/farmacologia , Diferenciação Celular/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citarabina/farmacologia , Relação Dose-Resposta a Droga , Citometria de Fluxo , Humanos , Células K562
15.
Chin Med J (Engl) ; 121(8): 691-5, 2008 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-18701020

RESUMO

BACKGROUND: This prospective, randomized, controlled study was designed to investigate the effects of a diabetes specific formula (Diason low energy: 313.8 kJ/100 ml), compared with a standard formula, on insulin sensitivity, serum C peptide, serum lipids and free fatty acid (FFA) in type 2 diabetics. METHODS: In total of 71 type 2 diabetics completed the study. Enteral formulas were given orally as the sole source of nutrition to the subjects for 6 days. Venous blood samples (0.5, 1, 2, 3 hours) were collected at day-7 after a 75 g oral glucose tolerance test (OGTT), day 1 after a standard test meal (1673.6 kJ) and after 6 days of either the test diabetes specific formula or a standard formula. Plasma glucose, serum insulin, C peptide and lipids were measured. RESULTS: After the intervention period, the diabetes specific formula resulted in a significantly lower postprandial rise in blood glucose concentrations at 0.5 hour (P < 0.05) and 1 hour (P < 0.01); significantly lower peak height of plasma glucose (P = 0.05); significantly lower plasma insulin concentrations at 0.5 hour (P < 0.01), 1 hour (P < 0.01) and 2 hours (P < 0.01); and a significantly lower plasma insulin peak compared to controls; both OGTT and a standard test meal (P < 0.05). The glucose and insulin area under the curve after the diabetes specific formula compared to the standard formula were significantly lower. The C peptide level was lower after 6 days of both nutrition formulas compare to 75 g OGTT, but not different from the standard mixed meal. Both formulas were well tolerated. CONCLUSIONS: In summary the diabetes specific formula with a relatively high monounsaturated fatty acid and high multi fiber proportion significantly improved glycemic control. On top of this, the insulin sensitivity (HOMA-IS) was significantly improved and may therefore directly improve the impact on long term complications. The disease specific formula should therefore be the preferred option to be used by diabetic and hyperglycemic patients in need of nutritional support.


Assuntos
Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/dietoterapia , Dieta para Diabéticos , Ácidos Graxos não Esterificados/sangue , Insulina/metabolismo , Peptídeo C/sangue , Humanos , Secreção de Insulina , Lipídeos/sangue , Pessoa de Meia-Idade , Estudos Prospectivos
16.
Front Pharmacol ; 9: 687, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30013477

RESUMO

Acute myeloid leukemia is an aggressive disease characterized by clonal proliferation and differentiation into immature hematopoietic cells of dysfunctional myeloid precursors. Accumulating evidence shows that CD34+CD38- leukemia stem cells (LSCs) are responsible for drug resistance, metastasis, and relapse of leukemia. In this study, we found that Nanog, a transcription factor in stem cells, is significantly overexpressed in CD34+ populations from patients with acute myeloid leukemia and in LSCs from leukemia cell lines. Our data demonstrate that the knockdown of Nanog inhibited proliferation and induced cell cycle arrest and cell apoptosis. Moreover, Nanog silencing suppressed the leukemogenesis of LSCs in mice. In addition, we found that these functions of Nanog were regulated by the insulin-like growth factor receptor (IGF1R) signaling pathway. Nanog overexpression rescued the colony formation ability of LSCs treated with picropodophyllin (PPP), an IGF1R inhibitor. By contrast, knockdown of Nanog abolished the effects of IGF2 on the colony formation ability of these LSCs. These findings suggest that the IGF2/IGF1R/Nanog signaling pathway plays a critical role in LSC proliferation.

17.
J Cancer ; 9(12): 2203-2210, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29937940

RESUMO

Background: Most hepatocellular carcinoma (HCC) patients have undergone a progression from chronic hepatitis, then liver cirrhosis (LC), and finally to carcinoma. The objective of this study was to elucidate risk factors to predict HCC development for cirrhosis patients. Methods: Multiple methylated specific PCR (MSP) was applied to determine methylation status of heparocarcinogenesis-related genes in 396 tissue and plasma specimens and multivariate cox model was used to analyze the relationship between risk variables and HCC development among cirrhosis patients, followed up in a median period of 30 months. Results: Among 105 LC cases, HCC incidence rate at 30 months was 30.48% (32/105), which were statistically associated with patients' age and aberrant methylation of p16, SFRP, and LINE1 (p<0.05). Receiver operating characteristic (ROC) curve showed the overall predictive accuracy reached the highest (90.7%) if the four risk variables were concurrent to predict HCC development. Moreover, along with the growth of age from 0-40, 40-55, to 55-70 years or the increased number of aberrantly-methylated gene from 0-1 to 2-3, the HCC incidence rate of cirrhosis patients rised from 10.00%, 12.28% to 82.14% and 17.44% to 89.47%, separately. Thus, based on combined analysis with diverse age and number of aberrantly-methylated gene, 105 cases were divided into five groups and computed their respective HCC incidecne rate to categorize them into different risk groups. Of note, A significant lifting of HCC incidence rate in the high-risk group (40-55 years coupled with 2-3 aberrantly-methylated genes, 55-70 years coupled with 0-1 aberrantly-methylated gene, 55-70 years coupled with 2-3 aberrantly-methylated genes; n=33) was observed compared with the low-risk group (0-40 years coupled with 0-1 aberrantly-methylated gene, 40-55 years coupled with 0-1 aberrantly-methylated gene; (n=72) (p<0.01). Conclusions: Ultimately, high-risk cirrhosis patients with 55-over years or 2-3 aberrantly-methylated genes should be paid more attention to be regularly screened with HCC development.

18.
Zhonghua Nei Ke Za Zhi ; 46(3): 193-6, 2007 Mar.
Artigo em Zh | MEDLINE | ID: mdl-17547798

RESUMO

OBJECTIVE: To investigate the effect of a new enteral nutrition suspension (diabetes) (TPF-DM) (Dixson 0.75 kcal/ml) (1 kcal = 4.184 kJ) on blood glucose, serum insulin and lipids as compared with a standard formula (Nutrition MF 0.75 kcal/ml) in patients with type 2 diabetes. METHODS: A randomized, controlled, paralleled and single center trial was carried out. A total of 76 patients with type 2 diabetes without using insulin and obvious complications were randomized into a study group and a control group. 36 patients in the study group and 35 in the control group completed the trial. The observation lasted 6 days. All calories came from enteral nutrition. At baseline all the patients had standard mixed meal (bread 50 g, egg 50 g, milk 250 ml, total calorie 400 kcal) test and at the end of the trial a enteral nutrient meal (enteral nutrient 400 ml, total calorie 300 kcal) test. Blood samples were taken before the meal and 30, 60, 120 and 180 minutes after the meal to test plasma glucose, serum insulin, serum lipids and some safety parameters. The area under curve (AUC) for plasma glucose, serum insulin, serum lipids was calculated. RESULTS: Compared with the mixed meal test, the AUC of plasma glucose and serum insulin during both Dixson 0.75 kcal/ml test and standard formula (Nutrition MF 0.75 kcal/ml) test were significantly lower (P < 0.01). The change at baseline in the study group was more than that in the control group [the change of AUC for plasma glucose (-6.42 +/- 8.62) h x mmol x L(-1) vs (-1.87 +/- 5.30) h x mmol x L(-1), P < 0.01; that of AUC for serum insulin (-36.94 +/- 49.77) h x mIU x L(-1) vs (-18.20 +/- 32.62) h x mIU x L(-1), P < 0.05]. Both the enteral nutrition formula can reduce insulin resistance (calculated by HOMA-IR), but there was no difference between them. There was no significant effect on total cholesterol, high density lipoprotein and low density lipoprotein. AUC of serum triglycerides was lower during the tests with both enteral nutrients than that during mixed meal test, but there was no significant difference between the two groups. There was no safety concern about the enteral nutrition. CONCLUSION: Enteral nutrition suspension (diabetes) (TPF-DM) (Dixson 0.75 kcal/ml) is an effective and safe enteral nutrient to be used in patients with type 2 diabetes.


Assuntos
Glicemia/metabolismo , Diabetes Mellitus Tipo 2/terapia , Nutrição Enteral , Diabetes Mellitus Tipo 2/sangue , Feminino , Humanos , Insulina/sangue , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Suspensões
19.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 29(3): 370-3, 2007 Jun.
Artigo em Zh | MEDLINE | ID: mdl-17633464

RESUMO

OBJECTIVE: To explore whether A1168C polymorphisms in paired box gene 4 (PAX4) are associated with type 1 diabetes mellitus (T1DM) in Chinese Han population. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype A1168C polymorphisms in PAX4 gene. Totally 109 patients with T1DM and 251 control subjects were recruited. The frequency distributions of genotypes between two groups were analyzed by SPSS software. RESULTS: The genotype distributions were in Hardy-Weinberg equilibrium both among T1DM cases and control subjects. No difference was observed in the genotype frequencies and allele frequencies between T1DM cases and control subjects (P > 0.05), nor was any disease association detected when patients were stratified according to age at diagnosis or sex (P > 0.05). CONCLUSION: The A1168C single nucleotide polymorphism in PAX4 gene may not play an essential role in genetic T1DM susceptibility in Chinese Han population.


Assuntos
Povo Asiático , Diabetes Mellitus Tipo 1/genética , Proteínas de Homeodomínio/genética , Fatores de Transcrição Box Pareados/genética , Estudos de Casos e Controles , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Polimorfismo de Nucleotídeo Único
20.
Oncol Rep ; 38(3): 1517-1524, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28713903

RESUMO

Heat-shock protein 90 (Hsp 90) acts as a molecular chaperone that maintains protein stability and regulates cell proliferation, survival, differentiation and apoptosis. The present study investigated the effect of Hsp90 inhibition on human acute myeloid leukemia (AML) cells using the novel small-molecule inhibitor SNX-2112. We found that SNX-2112 more potently inhibited KG-1a cell growth than the classical Hsp90 inhibitor 17-(2-dimethylaminoethyl)amino­17-demethoxygeldanamycin as determined by CCK-8 assay. Flow cytometry was used to examine the cell cycle, differentiation, and apoptosis, and western blotting and qRT-PCR were used to analyze the underlying mechanism. The results revealed that low concentrations of SNX-2112 arrested the cells in the G2/M phase and induced their differentiation and apoptosis, possibly by suppressing Akt and inhibitor of κB kinase, a component of the nuclear factor (NF)-κB signaling pathway. We also found that SNX-2112 increased the expression of the differentiation transcription factors PU.1 and CCAAT­enhancer-binding protein-α. Thus, SNX-2112 induced KG-1a cell differentiation, cell cycle arrest and apoptosis via modulation of Akt and NF-κB signaling, suggesting that it is a promising therapeutic agent for the treatment of AML.


Assuntos
Proteínas de Choque Térmico HSP90/antagonistas & inibidores , Compostos Heterocíclicos de 4 ou mais Anéis/administração & dosagem , Leucemia Mieloide Aguda/tratamento farmacológico , Proteínas Proto-Oncogênicas c-akt/genética , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proteínas de Choque Térmico HSP90/genética , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patologia , NF-kappa B/genética , Transdução de Sinais/efeitos dos fármacos
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