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1.
Chemosphere ; 238: 124648, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31524610

RESUMO

Deoxynivalenol (DON) is one of the most globally prevalent mycotoxins mainly produced by Fusarium species. It can cause pollution to water environmental quality due to its water solubility. Therefore, it is necessary to develop a green and efficient detoxification technology for DON. More importantly, the toxicity of the degradation products should be assessed. Photocatalytic degradation technology has attracted increasing attention in the field of pollutants treatment, especially for wastewater treatment. Herein, the as-prepared NaYF4:Yb,Tm@TiO2 composite (UCNP@TiO2) was employed as a novel photocatalyst for the NIR-enhanced photocatalytic degradation of DON. Three intermediate products were identified by using the ESI/MS analysis and secondary mass spectrogram, with the m/z values of 329.399, 311.243 and 280.913, respectively. Furthermore, the in vitro safety of the product mixtures with various degradation time (30 min, 60 min, 90 min and 120 min) were evaluated through the influences on cell viability, cell morphology, cell cycle, intracellular reactive oxygen species (ROS) level, cell apoptosis and antioxidant capacity of HepG2 cells. There were no significant differences in these investigated indicators between the control (free of DON) and 120 min products treatment. Overall, the results indicated that the toxicity of degradation products after 120 min irradiation was much lower and even nontoxic than that of DON.


Assuntos
Micotoxinas/química , Titânio/química , Tricotecenos/química , Purificação da Água/métodos , Sobrevivência Celular/efeitos dos fármacos , Células Hep G2/citologia , Células Hep G2/efeitos dos fármacos , Células Hep G2/metabolismo , Humanos , Micotoxinas/análise , Nanopartículas/química , Espécies Reativas de Oxigênio/metabolismo , Tricotecenos/toxicidade , Águas Residuárias/análise , Águas Residuárias/química , Poluentes da Água/química
2.
J Enzyme Inhib Med Chem ; 35(1): 152-164, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31742469

RESUMO

Overexpression of protein tyrosine phosphatase 1B (PTP1B) induces insulin resistance in various basic and clinical research. In our previous work, a synthetic oleanolic acid (OA) derivative C10a with PTP1B inhibitory activity has been reported. However, C10a has some pharmacological defects and cytotoxicity. Herein, a structure-based drug design approach was used based on the structure of C10a to elaborate the smaller tricyclic core. A series of tricyclic derivatives were synthesised and the compounds 15, 28 and 34 exhibited the most PTP1B enzymatic inhibitory potency. In the insulin-resistant human hepatoma HepG2 cells, compound 25 with the moderate PTP1B inhibition and preferable pharmaceutical properties can significantly increase insulin-stimulated glucose uptake and showed the insulin resistance ameliorating effect. Moreover, 25 showed the improved in vivo antihyperglycaemic potential in the nicotinamide-streptozotocin-induced T2D. Our study demonstrated that these tricyclic derivatives with improved molecular architectures and antihyperglycaemic activity could be developed in the treatment of T2D.


Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Desenho de Drogas , Inibidores Enzimáticos/farmacologia , Hipoglicemiantes/farmacologia , Proteína Tirosina Fosfatase não Receptora Tipo 1/antagonistas & inibidores , Terpenos/farmacologia , Animais , Glicemia/efeitos dos fármacos , Diabetes Mellitus Tipo 2/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Células Hep G2 , Humanos , Hipoglicemiantes/síntese química , Hipoglicemiantes/química , Resistência à Insulina , Masculino , Camundongos , Camundongos Endogâmicos , Estrutura Molecular , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Relação Estrutura-Atividade , Terpenos/síntese química , Terpenos/química
3.
Chemosphere ; 240: 124942, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31574434

RESUMO

Advanced oxidation processes (AOPs) can degrade heavy metal complexes in wastewater to improve the removal efficiency of metals. However, the influences of AOP treatments on toxicity induced by metal complexes are not well understood. This study compared the toxicity induced by EDTA-copper (Cu) after UV/persulfate (PS) and UV/H2O2 treatments on luminescent bacteria and human HepG2 cells. The results showed that EDTA-Cu complexes decreased Cu toxicity in luminescent bacteria but increased the cytotoxicity in HepG2 cells, indicating species-specific toxicity. The UV/PS and UV/H2O2 treatments under most pH values and [oxidant]/[EDTA-Cu] conditions decreased the toxicity of EDTA-Cu in HepG2 cells but increased the toxicity in luminescent bacteria. When the ratio of [oxidant] to [EDTA-Cu] was 10, low toxicity in treated solutions was observed in both UV treatment processes. The alkaline precipitation treatment had a significant influence on toxicity reduction after UV/PS treatment; however, it had minimal influence on the UV/H2O2 treatment system. The Cu and total organic carbon (TOC) removal efficiency cannot completely explain the results of toxicity assays. EDTA-Cu intermediates might play important roles in changing the toxicity of EDTA-Cu after both UV treatments. This study provides insights into evaluating the treatment efficiency of UV/PS and UV/H2O2 on EDTA-Cu decomplexation.


Assuntos
Complexos de Coordenação/toxicidade , Cobre/toxicidade , Ácido Edético/toxicidade , Peróxido de Hidrogênio/química , Photobacterium/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Transportadores de Cassetes de Ligação de ATP/metabolismo , Linhagem Celular Tumoral , Ácido Edético/química , Células Hep G2 , Humanos , Oxirredução , Raios Ultravioleta , Águas Residuárias/química
4.
J Sci Food Agric ; 100(1): 59-73, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31435933

RESUMO

BACKGROUND: Jiuzao is the residue after Bajiu distillation which is usually used as forage for livestock. However, it is not fully utilized yet considering the content of protein remained. The present study aimed to isolate antioxidant peptides from Jiuzao protein hydrolysates, then add these peptides into Baijiu product to enhance the healthy value of Baiju. Meanwhile environmental pollution caused by massive Jiuzao can be mitigated indirectly. RESULTS: Four peptides Ala-Tyr-Ile(Leu) (AYI(L)) and Asp-Arg-Glu-Ile(Leu) (DREI(L)) were identified from Jiuzao protein hydrolysates, the extraction contents of AYI + AYL and DREI + DREL were 896.10 and 110.51 mg kg-1 Jiuzao, respectively. On the one hand, antioxidant activities of these peptides were investigated. For in vitro antioxidant assays, AYI, AYL and DREI exhibited strong capacities in oxygen radical absorbance capacity (ORAC) assay. Furthermore, three levels of four peptides were assessed by 2,2'-azobis(2-methylpropanimidamidine) (AAPH)-induced HepG2 cells model. The results showed that these peptides exerted a degree of antioxidant activities in cells. Meanwhile, selected peptides concentrations according to cell assays remined at effective doses after in vitro digestion. On the other hand, the influence of these four peptides on the characteristic aroma compounds in Baijiu was studied. Most characteristic aroma compounds releases were increased with the addition of peptides. CONCLUSION: In the study, antioxidant activities of peptides were evaluated, the feasibility of utilizing Jiuzao protein hydrolysates to obtain beneficial peptides was also proved. Healthy effect of Baijiu or other food can be increased by adding these functional substances. The findings might contribute to food application and Baijiu industries. © 2019 Society of Chemical Industry.


Assuntos
Antioxidantes/química , Peptídeos/química , Hidrolisados de Proteína/química , Resíduos/análise , Vinho/análise , Antioxidantes/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Hep G2 , Humanos , Peptídeos/farmacologia , Hidrolisados de Proteína/farmacologia
5.
Food Chem ; 307: 125515, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31648177

RESUMO

This study evaluated the polyphenol profile and the antioxidative properties of Plinia trunciflora (O. Berg) Kausel fruits. Folin-Ciocalteau and pH-jumping methods indicated that these berries are a major source of antioxidant polyphenols (1201.05 mg GAE/100 g FW), particularly anthocyanins. HPLC-DAD-ESI-MS/MS analysis identified cyanidine glycosides as the main components. Flavon-3-ols and hydrolysable-tannins were also found. CAA assay showed that extracts of P. trunciflora fruits prevent lipid peroxidation in HepG2 cells with higher efficacy than other colourful fruits (CAA50 935.25 mg FW/mL cell medium). Moreover, our results suggested that the observed antioxidant protection involve both redox active properties of P. trunciflora components, as measured by ABTS, DPPH and FRAP assays, and upregulation of the genes coding for the antioxidant enzymes MnSOD and GPx, as evaluated by qRT-PCR. Collectively, our data provided evidence on the potential of P. trunciflora fruit as a very rich source of natural antioxidant molecules.


Assuntos
Antioxidantes/metabolismo , Suplementos Nutricionais/análise , Myrtaceae/química , Compostos Fitoquímicos/química , Cromatografia Líquida de Alta Pressão , Frutas/química , Frutas/metabolismo , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Células Hep G2 , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Myrtaceae/metabolismo , Compostos Fitoquímicos/análise , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Polifenóis/análise , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Espectrometria de Massas em Tandem , Regulação para Cima/efeitos dos fármacos
6.
Zhongguo Zhong Yao Za Zhi ; 44(20): 4481-4485, 2019 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-31872636

RESUMO

Aromatic constituents from rhizomes of Sophora tonkinensis were purified by extensive chromatographic techniques including column chromatography over macroporous resin,MCI,silica gel,weak acid cation exchange resin,Sephadex LH-20,ODS,and semi-preparative HPLC. Twelve aromatic compounds were isolated and identified from the water aqueous extract of the rhizomes of S.tonkinensis. Their structures were elucidated as 4-( 3-hydroxypropyl) phenol( 1),( ±)-4-( 2-hydroxypropyl) phenol( 2),benzamide( 3),( ±)-3-( p-methoxyphenyl)-1,2-propanediol( 4),4-methoxybenzamide( 5),3-hydroxy-1-( 4-hydroxy-3-methoxyphenyl) propan-1-one( 6),tyrosol( 7),( ±)-2,3-dihydroxypropyl benzoate( 8),vanillin alcohol( 9),7,3'-dihydroxy-8,4'-dimethoxyisoflavone( 10),7,4'-dihydroxy-3'-methoxyisoflavone( 11),and 7,3'-dihydroxy-5'-methoxyisoflavone( 12). Compounds 1-9 were firstly isolated from the Sophora genus. Compounds 4,5,10 and 11 can remarkably protect Hep G2 cell against APAP-induced damage at the concentration of 10 µmol·L-1. Compounds 1-12 exhibited no significant activities on the assays of inhibition of LPS-induced NO production in RAW cell lines and NF-κB inhibition.


Assuntos
Rizoma/química , Sophora/química , Cromatografia Líquida de Alta Pressão , Células Hep G2 , Humanos
7.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(11): 986-991, 2019 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-31878994

RESUMO

Objective To study the regulative effect of tumor protein p53 binding protein 2/p53 apoptosis-stimulating protein 2 (TP53BP2/ASPP2) on the autophagy of HepG2 human hepatoma cells and its mechanism. Methods The expression of ASPP2 was up-regulated or down-regulated by HepG2 cells infected with adenovirus and lentivirus. HepG2 cells were cultured in medium without fetal bovine serum for 24 hours. Western blotting was used to detect the expression of microtubule-associated protein 1 light chain 3 (LC3), beclin1, P62, autophagy-related gene 5 (ATG5), ATG7, and mammalian target of rapamycin (mTOR) pathway-associated protein mTOR, phospho-mTOR (p-mTOR), eukaryotic transcription initiation factor 4E binding protein 1 (4EBP1), phospho-4EBP1 (p-4EBP1), ribosomal protein S6, phospho-S6 (p-S6), ribosomal S6 kinase B1 (RPS6KB1/p70S6K), phospho-p70S6K (p-p70S6K). Fluorescence microscopy was used to detect the green fluorescent protein-microtubule-associated protein 1 light chain 3 (GFP-LC3) fusion protein expressed by the cells. At the same time, the Hep3B cells without p53 expression were infected with lentivirus, and the levels of ASPP2 in the cells were knocked down. Western blotting was performed to detect the expression of ASPP2 and mTOR pathway-associated protein mTOR, p-mTOR, 4EBP1, p-4EBP1, S6, p-S6, RPS6KB1/p70S6K, p-p70S6K. Results When the expression of ASPP2 was up-regulated, the mTOR complex 1 (mTORC1) pathway was activated, and the expression of autophagy-related proteins and the number of autophagosomes were reduced. After the expression of ASPP2 was down-regulated, the mTORC1 pathway was inhibited, and the expression level of autophagy-related proteins and the number of autophagosomes increased. In Hep3B cells with p53 expression silence, the mTORC1 pathway was still inhibited when ASPP2 wasdown-regulated. Conclusion ASPP2 inhibits the autophagy of HepG2 cells by activating mTOR pathway in a p53-independent manner.


Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Autofagia , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Células Hep G2 , Humanos , Proteína Supressora de Tumor p53
8.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(9): 843-848, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31750829

RESUMO

Objective To prepare polyclonal antibodies against Shisa like 1 protein (SSL1) and study the localization of SSL1 in hepatocellular carcinoma SMMC-7721 cells. Methods Human SSL1 gene was cloned from HepG2 cells by reverse transcription PCR, and then inserted into prokaryotic expression vector pET-28a to generate the SSL1 expression vector. The recombinant plasmid pET28a-SSL1 was then transformed into E. coli BL21 (DE3) and induced to express by IPTG. Polyclonal antibody against SSL1 was generated by immunizing Kunming mouse with the purified protein by the routine method. The specificity of polyclonal antibody was verified by Western blot analysis. The expression of SSL1 in SMMC-7721 cells was detected by immunofluorescent cytochemistry. Golgi complexes were signed by Golgi-Tracker Red to analyze the subcellular localization of SSL1 protein in SMMC-7721 cells. Results The SSL1 gene was cloned and the recombinant vector pET28a-SSL1 was successfully constructed. Pure SSL1 protein expression in E. coli BL21 was confirmed and polyclonal antibodies against protein SSL1 was obtained in immunized Kunming mice. Immunofluorescent cytochemistry showed that SSL1 was expressed in the cytoplasm, and was co-localized with Golgi-Tracker Red in SMMC-7721 cells. Conclusion We have obtained SSL1 polyclonal antibodies with high specificity, which was proved situated in Golgi bodies of SMMC-7721 cells.


Assuntos
Anticorpos , Proteínas de Membrana/imunologia , Animais , Especificidade de Anticorpos , Western Blotting , Escherichia coli , Complexo de Golgi , Células Hep G2 , Humanos , Camundongos , Plasmídeos
9.
J Environ Pathol Toxicol Oncol ; 38(2): 119-131, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31679275

RESUMO

BACKGROUND/AIMS: LncRNAs are significant regulators in multiple cancers including hepatocellular carcinoma (HCC). Recently, lncRNA ANRIL has been reported to be elevated during multiple cancer types, exhibiting oncogenic roles. However, the exact biological mechanism of ANRIL is still poorly understood in HCC. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) assays were utilized to detect expressions of ANRIL, miR-384, and STAT3. CCK8 and EDU assays were employed to evaluate HCC cell proliferation. A flow cytometry assay was used to detect the HCC cell cycle and cell apoptosis. The scratch migration and Transwell invasion assays were performed to test cell migration and invasion, respectively. RIP and RNA pull-down assays were carried out to confirm the correlation between ANRIL and miR-384. The dual-luciferase reporter assay was used to prove the association between miR-384 and STAT3. Western blotting analysis was performed to examine protein levels of STAT3. IHC and HE staining were employed to detect Ki-67 and histopathology. RESULTS: ANRIL expression was upregulated in HCC cells, including SMCC7721, HepG2, MHCC-97H, SNU449 and HUH-7 cells, in comparison to the normal human liver cells LO2. Knockdown of ANRIL suppressed HCC cell proliferation and induced cell cycle arrest and apoptosis. HCC cell migration and invasion capacity were inhibited by inhibition of ANRIL. Bioinformatics analyses revealed that ANRIL could interact with miR-384. miR-384 was significantly decreased in HCC cells, and overexpression of miR-384 repressed HCC progression. STAT3 was predicted as a target of miR-384, and miR-384 can modulate STAT3 levels negatively in vitro. ANRIL can suppress HCC development through regulating miR-384 and STAT3 in vivo. CONCLUSION: ANRIL is involved in HCC progression by direct targeting of miR-384 and STAT3. Also, ANRIL could act as a potential candidate for HCC diagnosis, prognosis, and therapy.


Assuntos
Carcinogênese/genética , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , MicroRNAs/genética , RNA Longo não Codificante/genética , Fator de Transcrição STAT3/genética , Carcinoma Hepatocelular/fisiopatologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Progressão da Doença , Células HEK293 , Células Hep G2 , Humanos , Neoplasias Hepáticas/fisiopatologia , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Fator de Transcrição STAT3/metabolismo
10.
J Environ Pathol Toxicol Oncol ; 38(3): 195-203, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31679307

RESUMO

UNCI 19 expression has been reported to be significantly higher in hepatic cancer cells (HCC). However, the clinical significance of modulating UNC119 expression in HCC is not well understood. The study described here aimed to explore the potential of curcumin in modulation of UNC119 expression in HCC by assessment with quantitative real-time PCR, western blot, and immune-histochemical analyses in HCC cell lines and tissues. The biological functions of UNC119 in the proliferation, growth, and cycle of tumor cells were analyzed both in vitro and in vivo. UNC119 expression was upregulated in HCC cell lines and tissues as indicated by comparison with normal liver cells and tissues. Cellular function assays showed that higher levels of UNC119 not only promoted proliferation but also enhanced HCC cell migration and invasion. UNC119 promoted progression of the cell cycle and significantly promoted HCC cell growth through the Wnt/ß-catenin signal pathway, and enhanced tumor migration and invasion by the TGF-ß/EMT pathway. Curcumin efficiently inhibited HCC cell proliferation by blocking the Wnt/ß-catenin pathway and inhabited migration and invasion by blocking the TGF-p/EMT signal pathway. Curcumin not only was beneficial for tumor remission but also contributed to the long-term survival of HCC-bearing mice. UNC119 was significantly upregulated and promoted cell growth in hepatic cancer cells and tissues by the Wnt/ß-catenin signal pathway and migration by TGF-ß/EMT signal pathway. Curcumin treatment inhibited cell proliferation, growth, migration, and invasion by inhibition of those pathways.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Antineoplásicos/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Curcumina/farmacologia , Invasividade Neoplásica/genética , Metástase Neoplásica/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Linhagem Celular Tumoral , Movimento Celular/genética , Feminino , Células Hep G2 , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Organismos Livres de Patógenos Específicos
12.
Tumour Biol ; 41(10): 1010428319880080, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31603389

RESUMO

Searching for new sources of safe nutraceuticals antitumor drugs is an important issue. Consequentially, this study designed to assess the antitumor activity of Pulicaria undulata extract in vitro in the treatment of hepatocellular carcinoma HepG2 cell line. Aerial parts of P. undulata plants were collected, used for phytochemical analysis, and assessed for anticancer activity. The antitumor activity was evaluated through studying the cell viability and apoptotic pathway. The gas chromatography-mass spectrometry phytochemical analysis revealed that P. undulata is a promising new source of several known antioxidant and antitumor compounds which could participate in drug development and exploration of alternative strategies to the harmful synthetic antitumor drugs. P. undulata stifled HepG2 cell viability in a concentration-dependent manner. Meanwhile, P. undulata tempted substantial apoptosis in HepG2 cells and enhanced the expression of miR-34a. However, the mRNA expression level of antiapoptotic B-cell lymphoma-2 was markedly decreased by P. undulata treatment. Moreover, P. undulata increased the protein expression of proapoptotic p53 and caspase 3/9 with reducing B-cell lymphoma-2 protein expression level. Thus, P. undulata induced apoptosis in the HepG2 cells by overexpression of miR-34a which regulates p53/B-cell lymphoma-2/caspases signaling pathway. These findings were well appreciated with morphological studies of cells treated with P. undulata. In conclusion, P. undulata could be a probable candidate agent for the initiation of cell apoptosis in HepG2 and thereby can serve as promising therapeutic agent for treatment of hepatocellular carcinoma which should attract further studies.


Assuntos
Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/patologia , Proliferação de Células/efeitos dos fármacos , Neoplasias Hepáticas/patologia , Extratos Vegetais/farmacologia , Pulicaria/química , Carcinoma Hepatocelular/tratamento farmacológico , Células Hep G2 , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Transdução de Sinais
13.
Zhonghua Gan Zang Bing Za Zhi ; 27(9): 687-692, 2019 Sep 20.
Artigo em Chinês | MEDLINE | ID: mdl-31594093

RESUMO

Objective: To construct the recombinant adenoviral containing fructose 1, 6-biphosphatase 1 (FBP1), and to investigate whether FBP1 has effect on autophagy and proliferation in liver cancer cells (HepG2). Methods: FBP1 cDNA sequence was amplified by PCR and cloned in adenovirus vector pAdTrack-TO4, and then recombinant adenovirus plasmid pAdTrack-FBP1 was constructed. The recombinant adenovirus plasmid was transfected into HEK293 cells by Lipofectamine 3000. High-titer of recombinant adenovirus AdFBP1 was obtained by packaging and amplification. HepG2 cells were infected with recombinant adenovirus AdFBP1, and the Mock and AdGFP group were set at the same time. Western blot and confocal laser scanning microscopy were used to observe the effect of FBP1 on the level of autophagy in hepatocellular carcinoma cells, and the effect of FBP1on the proliferation was observed by MTS and colony formation assay. A t-test and one-way ANOVA were used to compare the mean between group. Results: A high-titer recombinant adenovirus FBP1 was successfully constructed. Western blot and confocal laser scanning microscopy showed that the level of autophagy in AdFBP1 group was significantly lower than that in AdGFP group. Western blot results showed that LC3-II protein expression level in AdGFP was 1.10 ± 0.10 and 0.30 ± 0.01 in AdFBP1 group, F = 90.36, P < 0.01. Confocal laser scanning microscopy analysis showed that the average number of autophages in AdGFP was 28.33 ± 1.53 and 12.33 ± 1.53 in AdFBP1group, F = 97.40, P < 0.01. In addition, the results of colony formation assay and MTS assay showed that the proliferation of liver cancer cells in the AdFBP1 group was significantly inhibited compared with the AdGFP group. The results of colony formation showed that the cell clones in the AdGFP group was 65.66 ± 2.57 and 34.00 ± 2.00 in AdFBP1 group, F = 141.50, P < 0.01. MTS results showed that the absorbance of AdGFP group at 96h was 39.13 ± 2.21 and 30.61 ± 3.33 in AdFBP1 group, F = 7.80, P < 0.05. Conclusion: FBP1 inhibited the autophagy and proliferation in liver cancer cells (HepG2).


Assuntos
Autofagia , Proliferação de Células , Frutose-Bifosfatase/metabolismo , Neoplasias Hepáticas/patologia , Adenoviridae , Vetores Genéticos , Células HEK293 , Células Hep G2 , Humanos , Neoplasias Hepáticas/enzimologia , Transfecção
14.
Zhonghua Gan Zang Bing Za Zhi ; 27(9): 693-697, 2019 Sep 20.
Artigo em Chinês | MEDLINE | ID: mdl-31594094

RESUMO

Objective: To investigate the effect and mechanism of XTP4 gene in apoptotic hepatoblastoma HepG2 cell line. Methods: HepG2 cells were transiently transfected with small interfering RNA of XTP4 genes, plasmid pcDNA3.1/myc-His(-) A-XTP4, and hepatitis B virus X protein transactivated x gene 4 (HBX protein trans-activate gene4, XTP4) and their respective negative controls. After 48h, the overexpression and interference expression condition of XTP4 in HepG2 cells were detected by Western blot. HepG2 cells apoptosis was detected by flow cytometry. The expression levels of apoptosis-related proteins P53, Bcl-2, Bax and Caspase-3 in HepG2 cells were detected by Western blot, and Bcl-2/Bax ratio was calculated. The chemiluminescence assay was used to detect activity of caspase-3 in HepG2 cells. The measured data were presented as (x ± s), and independent sample t-test was used for comparison between the two groups. Results: HepG2 cells had successfully achieved the overexpression and interference expression of XTP4 protein. Compared with the control group, the overexpression of XTP4 in HepG2 cells had significantly decreased the number of apoptotic cells (P < 0.05), and increased Bcl-2/Bax (P < 0.05) ratio, but decreased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was decreased (P < 0.05). However, interference with XTP4 expression in HepG2 cells had significantly increased the number of apoptotic cells (P < 0.05) and decreased Bcl-2/Bax (P < 0.05) ratio, but increased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was increased (P<0.05). Conclusion: In HepG2 apoptosis XTP4 has inhibitory effect, and its effect on inhibiting HepG2 apoptosis may be achieved by regulating the Bcl-2/Bax ratio, and the P53 protein may be involved.


Assuntos
Apoptose , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Transativadores/metabolismo , Caspase 3/metabolismo , Células Hep G2 , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transfecção , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismo
15.
Anticancer Res ; 39(10): 5393-5401, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31570434

RESUMO

BACKGROUND/AIM: Local recurrence of hepatocellular carcinoma (HCC) after thermal coagulation therapy may be associated with an aggressive phenotypic change. This study focused on the thermal effects on HCC cells and evaluated the heat shock response and phenotypic changes after heat treatment. MATERIALS AND METHODS: HepG2 and HuH7 cells were used. After heat treatment at 37-50°C for 5-30 min, we assessed their survival rate, induction of heat shock protein (HSP)70 promoter, proliferation rate, induction of the epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC)-related markers. RESULTS: Induction of HSP70 promoter per surviving cell was maximized after 10 min of heat treatment at 48°C. Induction of EMT and CSC-related markers was also observed. CONCLUSION: Sub-lethal heat treatment causes large heat shock response to surviving HCC cells and induce EMT-like and CSC-like phenotypic changes that might contribute to increased aggressiveness.


Assuntos
Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Resposta ao Choque Térmico/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Transição Epitelial-Mesenquimal/genética , Proteínas de Choque Térmico HSP70/genética , Células Hep G2 , Temperatura Alta , Humanos , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/patologia , Células-Tronco Neoplásicas/patologia , Taxa de Sobrevida
16.
Chem Biol Interact ; 314: 108847, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31610155

RESUMO

Lead (Pb) is one of the toxic heavy metals that have several toxicological implications including cytotoxicities and oxidative stress. The release of reactive oxygen species (ROS) usually initiates lipid peroxidation and resulting in inflammation and tissue injury. However, the detailed identification of the Pb-produced lipid hydroperoxides has received little attention. Furthermore, the mechanisms behind such effects are less informed. Therefore, this study firstly investigated Pb-produced lipid hydroperoxides in human HepG2 cells using LC/MS. The effects of Pb on the antioxidant enzymes were additionally examined using qPCR and their dependent activities. As a protection trial, the ameliorative effects of rosmarinic (RMA) and ascorbic (ASA) acids on Pb-induced cytotoxicity and oxidative stress and their regulatory effects on Nrf2/Keap1 pathway were investigated. The achieved results confirmed cytotoxicity and oxidative damage of Pb on HepG2 cells. In addition, 20 lipid hydroperoxides (LOOH) were identified including 11 phosphatidylcholine hydroperoxides (PCOOH), 5 triacylglycerol hydroperoxides (TGOOH) and 4 cholesteryl ester hydroperoxides (CEOOH). The most dominant LOOH species were PCOOH 34:2, PCOOH 34:3, PCOOH 38:7, TGOOH 60:14, TGOOH 60:15, CEOOH 18:3 and CEOOH 20:4. Pb significantly downregulated Nrf2-regulated antioxidant enzymes at both the pretranscriptional and functional levels. Co-exposure of HepG2 cells to RMA and ASA significantly reduced Pb-produced adverse outcomes. This protection occurred via activation Nrf2-Keap1 antioxidant pathway.


Assuntos
Antioxidantes/metabolismo , Ácido Ascórbico/química , Cinamatos/química , Depsídeos/química , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Chumbo/química , Fator 2 Relacionado a NF-E2/metabolismo , Ácido Ascórbico/farmacologia , Cromatografia Líquida de Alta Pressão , Cinamatos/farmacologia , Depsídeos/farmacologia , Regulação para Baixo/efeitos dos fármacos , Células Hep G2 , Humanos , Chumbo/toxicidade , Peróxidos Lipídicos/análise , Peróxidos Lipídicos/metabolismo , Espectrometria de Massas , NAD(P)H Desidrogenase (Quinona)/genética , NAD(P)H Desidrogenase (Quinona)/metabolismo , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Superóxido Dismutase-1/genética , Superóxido Dismutase-1/metabolismo
17.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 50(4): 520-526, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31642229

RESUMO

OBJECTIVE: To investigate the effect of nuclear receptor Rev-erbß knockout on proliferation and migration ability of human hepatocellular carcinoma cell line HepG2. METHODS: -The Rev-erbß gene knockout HepG2 cell line was abtained by CRISPR/Cas9 genome editing technique with specific DNA modification of the target gene. The Rev-erbß gene targeting vectors were co-transfected into HepG2 cells. Through cloning and screening, the Rev-erbß gene knockout HepG2 cell line was constructed, PCR, sequencing and Western blot methods were carried out for the identification of the Rev-erbß gene knockout HepG2 cell line. The expression level of tumor migration and invasion-associated gene in Rev-erbß gene knockout cell was determined by real-time quantitative PCR (qRT-PCR) and was compared with normal cell as control.MTT, cell scratch and Transwell experiments were conducted in order to explore the effect of Rev-erbß gene on HepG2 cell's ability of proliferation, migration and invasion. RESULTS: A Rev-erbß gene knockout monoclonal cell line, which was identified by PCR, sequencing and Western blot, was successfully constructed and named HepG2 C5 (Rev-erbß -/-). qRT-PCR results showed that Rev-erbß knockout resulted in up-regulation of matrix metalloproteinase-2 (MMP2), matrix metalloproteinase-9 (MMP9) and extracellular matrix protein-1 (ECM1) gene expression (P < 0.05) and down-regulation of E-cadherin (CDH1) gene expression (P=0.05).Results of MTT, cell scratch and transwell experiments showed that HepG2 C5 had stronger proliferation, migration and invasion ability than control cells (P < 0.05). CONCLUSION: Rev-erbß gene knockout could change the expression of migration and adhesion-associated genes in HepG2 cell, and then affect the proliferation, migration and invasion ability of HepG2 cells.


Assuntos
Movimento Celular , Proliferação de Células , Invasividade Neoplásica , Receptores Citoplasmáticos e Nucleares/genética , Proteínas Repressoras/genética , Proteínas da Matriz Extracelular/metabolismo , Técnicas de Inativação de Genes , Células Hep G2 , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo
18.
Analyst ; 144(22): 6609-6616, 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31616873

RESUMO

Maintaining the redox balance of biological systems is a key point to maintain a healthy physiological environment. Excessive iron ions (Fe3+) can cause apoptosis, tissue damage and death. Fortunately, ascorbic acid (AA) as a reducing agent has been evaluated for the reduction of Fe3+. Moreover, AA plays an important role in relieving hypoxia-induced oxidative stress. Therefore, the real-time imaging of the Fe3+ and AA fluctuations is important for understanding their biofunctions in cells and in vivo. In this work, we developed a fluorescent nanoprobe carbon dot-desferrioxamine B (CD-DB) by the conjugate connection of CDs and desferrioxamine B (a complexing agent for Fe3+) for the associated detection of Fe3+ and AA. CD-DB exhibited excellent sensitivity and selectivity for the detection of Fe3+ and AA. The nanoprobe CDs-DB@Fe obtained by the reaction of CD-DB and Fe3+ was suitable for tracing the dynamic changes of AA in cells and in vivo. Therefore, CDs-DB@Fe was used for monitoring the fluctuation of AA in hypoxic cell models, hypoxic zebrafish models and liver ischemia mice models. These results exhibited the decrease in AA under hypoxic conditions because AA was consumed to neutralize free radicals and relieve hypoxia-induced oxidative stress damage. The ideal biocompatibility and low toxicity make our nanoprobe a potential candidate for the research of the physiological effects of AA in vivo.


Assuntos
Ácido Ascórbico/análise , Corantes Fluorescentes/química , Hipóxia/metabolismo , Ferro/análise , Pontos Quânticos/química , Animais , Carbono/química , Hipóxia Celular , Desferroxamina/química , Corantes Fluorescentes/síntese química , Células Hep G2 , Humanos , Isquemia/metabolismo , Limite de Detecção , Fígado/irrigação sanguínea , Fígado/metabolismo , Camundongos Endogâmicos BALB C , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Estresse Oxidativo , Peixe-Zebra
19.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 36(5): 803-809, 2019 Oct 25.
Artigo em Chinês | MEDLINE | ID: mdl-31631629

RESUMO

Cell freeze-drying can be divided into the freezing and drying processes. Mechanical damage caused by ice crystals and damage from solute during freezing shall not be ignored and lyoprotectants are commonly used to reduce those damages on cells. In order to study the mechanism of lyoprotectants to protect cells and determine an optimal lyoprotectant formula, the thermophysical properties and percentage of unfrozen water of different lyoprotectants in freezing were investigated with differential scanning calorimeter (DSC). The survival rate indicated by trypan blue exclusion test and cell-attachment rate after 24 h using different lyoprotectants to freeze hepatoma Hep-G 2 cells were measured after cell cryopreservation. The results show that 40% (W/V) PVP + 10% (V/V) glycerol + 15% (V/V) fetal bovine serum + 20% (W/V) trehalose formula of lyoprotectant demonstrate the best effect in protecting cells during freezing, for cell-attachment rate after 24 h is 44.56% ± 2.73%. In conclusion, the formula of lyoprotectant mentioned above can effectively protect cells.


Assuntos
Criopreservação , Crioprotetores/química , Congelamento , Trealose/química , Varredura Diferencial de Calorimetria , Liofilização , Células Hep G2 , Humanos
20.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 36(5): 850-855, 2019 Oct 25.
Artigo em Chinês | MEDLINE | ID: mdl-31631635

RESUMO

There is a great demand for blood and stem cells in clinic. It is difficult to achieve high throughput and to increase the cooling rate at the same time during vitrification. In this paper, a micro-droplet spray system with a container collection device was fabricated, and HepG2 cells were sprayed by this system for high-throughput vitrification. First, the container collection device and a cryo-paper were used to receive micro-droplets in the spray vitrification system. The results showed that the cell survival rate and 24h adhesion rate in container collection vitrification group were significantly higher than those in cryo-paper collection group. Second, HepG2 cells were sprayed and vitrified at increased cell density, and it was found that the results of micro-droplet spray vitrification did not change significantly. Finally, micro-droplet spray vitrification is compared with slow freezing. Cell processing capacity in the vitrification group increased, meanwhile, the cell survival rate and 24h adhesion rate in the vitrification group were significantly higher than those in slow freezing group. The results indicated that the micro-droplet spray vitrification system with container collection device designed in this paper can achieve high-throughput cell vitrification, which is of great significance for mass preservation of small cells.


Assuntos
Criopreservação , Vitrificação , Adesão Celular , Sobrevivência Celular , Células Hep G2 , Humanos
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