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Effects of the Interaction between Hydroxyapatite Nanoparticles and hepatoma cells

Journal of Wuhan University of Technology(Materials Science) 2014年第3期29卷 635-642页

作者：尹美珍 XU Weiguo CUI Bingcun 戴红莲 HAN Yingchao YIN Yixia LI Shipu School of Medicine Hubei Polytechnic University Tongji Hospital Tongji Medical College Huazhong University of Science and Technology State Key Laboratory of Advanced Technology for Materials Synthesis and Processing Wuhan University of Technology Biomedical Materials and Engineering Center Wuhan University of Technology

To gain a better understanding of the anticancer effects of hydroxyapatite （HAP） nanoparticles in vivo and in vitro, the effects of the interaction of HAP nanoparticles with hepatoma cells were explored. HAP nanoparticles were prepared by homogeneous precipitation and characterized by laser particle analysis and transmission electron microscopy （TEM）. HAP nanoparticles were observed to be uniformly distributed, with rod-like shapes and diameters in the range of 42.1-87.1 nm. Overnight attached, suspended, and proliferating Bel-7402 cells were incubated with HAP nanoparticles. Inverted microscopy observation revealed that HAP nanoparticles with a cell membrane showed good adsorption. TEM demonstrated that HAP nanoparticles were present on the surface of cells, continuously taken up by cells through endocytosis, and transported in vesicles close to the nucleus. Fluorescence microscopy showed that the concentrations of intracellular Ca2＋ labeled with Fluo-3 calcium fluorescent probe were significantly enhanced. In addition, inverted microscopy observation revealed that suspended cells treated with HAP nanoparticles did not adhere to the culture bottle, resulting in cell death. After the overnight attached cells were treated with HAP nanoparticles for 96 h with increasing doses of HAP nanoparticles, inverted microscopy observation revealed that cell proliferation was slowed and ceU-ceU adhesion was weakened. Feulgen staining and image analysis indicated that the nuclear DNA content of the cells was markedly reduced, and argyrophilic nucleolar organizer region （AgNOR） staining and image analysis indicated that the number of AgNORs was significantly decreased. Therefore, hepatoma cells brought about the adsorption, uptake, transport and degradation of HAP nanoparticles. In addition, HAP nanoparticles affected hepatoma cells with regard to cell-cell adhesion, cell and extracellular matrix adhesion, and DNA and protein synthesis; thus inhibiting cell proliferation. This understanding of the effects of interaction between

关键词： hydroxyapatite nanoparticles hepatoma cells interaction effects cell adhesion

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Differentiation of human hepatoma Hep G2 cells induced by 10-hydroxyca mptothecin

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Chinese Medical Journal 2000年第8期113卷 40-41页

作者：章雄文周燕华胥彬 Department of Pharmacology Shanghai Inst. of Material Medical Chinese Academy of Sciences Shanghai 200031 China

ObjectiveTo study the difterentiation-inducing effect of hydroxycamptothecin(HCPT)on hepatoma Hep G2 cells.Methods3-[4, 5-dimethylthial-2-yl]-3, 5-diphenyl-tetrazolinium bromide(MTT)colorimetric method was used for determining HepG2 cells viability. Alpha-fetoprotein(AFP)and albumin(ALB)levels were determined with Elisa Kits.ResultsThe AFP secretion of HCPT-treated cells was reduced by 44.0%-49.2% while the ALB level was increased markedly.ConclusionHydroxycamptothecin can induce human hepatoma Hep G2 cells to differentiation. Chin Med J 2000; 113(8): 712-713

关键词： differentiation hepatoma cells hydroxycamptothecin

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Dissection the endocytic routes of viral capsid proteins-coated upconversion nanoparticles by single-particle tracking

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Chinese Chemical Letters 2022年第10期33卷 4710-4714页

作者： Yujun Ning Lin Wei Shen Lin Yifan Jiang Naidong Wang Lehui Xiao College of Chemistry Zhengzhou UniversityZhengzhou 450001China State Key Laboratory of Medicinal Chemical Biology Tianjin Key Laboratory of Biosensing and Molecular RecognitionCollege of ChemistryNankai UniversityTianjin 300071China College of Chemistry and Chemical Engineering Hunan Normal UniversityChangsha 410081China College of Veterinary Medicine Hunan Agricultural UniversityChangsha 410128China School of Chemistry and Chemical Engineering Henan Normal UniversityXinxiang 453007China

Real-time exploring the cellular endocytic pathway of viral capsid proteins(VCPs)functionalized nanocargos at the single-particle level can provide deep insight into the kinetic information involved in virus infection.In this work,porcine circovirus type 2(PCV2)VCPs with different functions are modified onto the surface of upconversion nanoparticles(VCPs-UCNPs)to investigate the cellular internalization process in real-time.Clathrin-mediated endocytosis is found to be the essential uptake mechanism for these VCPs-UCNPs.Besides,it is verified that P_(1)-UCNPs(PCV2 VCPs with nuclear localization signal,namely P1)can be easily assembled close to the perinuclear area,which is different from that of P_(2)-UCNPs(PCV2 VCPs without nuclear localization signal,namely P_(2)).Interestingly,multistep entry processes are observed.Particularly,confined diffusion is observed during the transmembrane process.The intracellular transport of VCPs-UCNPs is dependent on microtubules toward the cell interior.During this process,P_(1)-UCNPs display increased velocities with active transport,while diffusion much faster around the perinuclear area.But for P_(2)-UCNPs,there are only two phases involved in their endocytosis process.This study presents distinct dynamic mechanisms for the nanocargos with different functions,which would make a useful contribution to the development of robust drug delivery systems.

关键词： Upconversion nanoparticles Viral capsid proteins Single-particle tracking hepatoma cells Endocytosis

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HBx induced AFP receptor expressed to activate PI3K/AKT signal to promote expression of Src in liver cells and hepatoma cells

HBx induced AFP receptor expressed to activate PI3K/AKT sign...

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第七届生命科学联合学术大会

作者： Mingyue Zhu Junli Guo Wei Li Hua Xia Yan Lu Xu Dong Yi Chen Xieju Xie Shigan Fu Mengsen Li Hainan Provincial Key Laboratory of Carcinogenesis and Intervention Hainan Medical College Key Laboratory of Molecular Biology Hainan Medical College Department of Pathophysiology Hainan Medical College Department of Physiology Hainan Medical College Institution of Tumor Hainan Medical College

Background Hepatitis B virus(HBV)-X protein(HBx)is a transactivator of host several cellular genes including alpha-fetoprotein(AFP)and AFP receptor(AFPR)which contributes to HBV-associated tumor development.The expression of AFP/AFPR are correlated with hepatocellular carcinoma(HCC)-initial cells.But the role of AFP and AFR in promoting occurrence of HBV-related HCC were still unclear.Methods A total of 71 clinical patients’liver specimens,normal human liver cells L-02 and HCC cell lines,PLC/PRF/5 were selected for analyzing the effects of HBx on expression of AFP,AFPR and Src.The expression of goal proteins were detected by Immunohistochemical stained and Western blotting;HBx-expressed vectors were constructed and transfected into L-02 cells,laser confocal microscopy was applied to observe expression and location of AFP,AFPR and Src in the normal liver cells and HCC cells,soft agar colony formation assay was used to observe colonies formed of the cells.Results We confirmed HBx gives preference to promote the expression of AFP and AFPR;HBx priors to up-regulate the expression of AFPR and AFP in L-02 cells and in normal liver specimens;AFPR signal been able to stimulate Src expressed.The results also indicated that phosphatidylinositol 3-kinase(PI3K)inhibitors Ly294002 and GDC0941 effectively suppress AFPR mediated up-regulation expression of Src in AFPR positive HCC lines.Conclusions HBx priors to drive the expression of AFP and AFPR to promote expression of Src in normal liver cells and hepatoma cells;AFP and AFPR maybe play pivotal role in HBV-related hepatocarcinogenesis;Targeting AFPR is an available therapeutic strategy of HCC.

关键词： Hepatitis B virus-x（HBx） Alpha fetoprotein（AFP）/AFP receptor PI3K/AKT signal hepatoma cells

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Cucurbitacin E inhibits the proliferation of hepatoma cells in vitro and in vivo through induction of G2/M phase arrest

Cucurbitacin E inhibits the proliferation of hepatoma cells ...

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两岸三地药理学与临床药理学术会议

作者： LI Yan-chun~1 MA En-long~1 DENG Yi-hui~2 JING Yong-kui~3 (1. Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang 110016, China 2. Department of Pharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, China 3. Department of Medicine, Mount Sinai School of Medicine, New York, USA)

Objective Cucurbitacins are the highly oxygenated tetracyclic triterpenes,which are predominant- ly found in the Cucurbitaceae family but are also present in several other families of the plant kingdom.A number of compounds of this group have been investigated for their cytotoxic,hepatoprotective,anti-inflam- matory,cardiovascular and anti-diabetic activities.In China,the cucurbitacin preparation,which contains mostly cucurbitacin B and cucurbitacin E,has been clinically used for the treatment of the primary liver car- cinoma.It has been previously reported that cucurbitacin E could produce cytotoxicity against a variety of cancer cells,and various mechanisms were implicated in its cytotoxic effect.The present study is to investi- gate the effect of cucurbitacin E on hepatoma cells in vitro and in vivo and to study their potential mecha- nisms of action.Methods The MTT assay was used to assess the viability of human HepG2 and BEL7402 hepatoma cells in vitro after treatment with different concentrations of cucurbitacin E.The cell cycle distri- bution was determined by flowcytometric analysis after propidium iodide(PI)staining.The cell cycle-relat- ed proteins were detected using western blotting analysis.Implanted mouse hepatoma H22 model was built to evaluate the growth inhibitory effect of cucurbitacin E in vivo in mice.Results Our studies found that cu- curbitacin E(10-300 nM)produced anti-proliferative effect on human HepG2 and BEL7402 hepatoma cells in vitro without cytotoxicity.According to flowcytometric analysis,cucurbitacin E arrested the cell cycle at G2/M phase in both HepG2 and BEL7402 hepatoma cells after 24 h treatment.Cucurbitacin E induced the decrease in the level of CDK1 protein and the increase in the level of p21 protein,but had no effect on the levels of cyclin A,cyclin B1 and Cdc25C protein.In in vivo anti-tumor experiment,cucurbitacin E had sig- nificant inhibitory effects on the growth of mouse H22 hepatoma cells.Conclusions Cucurbitacin E inhibited the proliferation of hepatoma cells in vi

关键词： cucurbitacin e hepatoma cells G2/M arrest

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Selective Anti-hepatoma Treated with Titanium Oxide Nanoparticles in vitro

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Journal of Wuhan University of Technology(Materials Science) 2003年第1期18卷 52-54页

作者：曹献英 WuhanUniversityofTechnology

Effects of titanium oxide (TiO 2) nanoparticles on Bel-7402 human hepatoma cells and L-02 human hepatocytes at different times were observed.Using cell culture,cell growth curves of Bel-7402 cells and L-02 cells treated with TiO 2 nanoparticles were examined by MTT assay,and the cellular ultrastructure was observed by an analytical transmission electron microscope (ATEM).It is found that OD value of Bel-7402 cell treated with TiO 2 nanoparticles for 48-144h is obviously lower than that of control group (pcells is almost not affected by TiO 2 nanoparticles.ATEM and energy dispersive X ray (EDX) analyses show that there are obvious vacuoles increased heterolysosome,and particles with high electron density which are confirmed to be TiO 2 nanoparticles in Bel-7402 cytoplasm.More interestingly,it is alse found that TiO 2 nanoparticle obviously inhibits the proliferation of hepatoma cells by altering lysosome activity and destroying cytoplasm structure.The inhibition on proliferation of hepatocytes by TiO 2 nanoparticles is much slighter.The results demonstrate that TiO 2 nanoparticle has different killing effects on cancer cell and normal cell.

关键词： titanium oxide nanoparticle hepatoma cells

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Protective effects of Amaranthus hybridus against aflatoxin B1 and fumonisin B1-induced genotoxicity in H4IIE-luc cells

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hepatoma Research 2015年第1期1卷 136-146页

作者： Mohamed I.M.Ibrahim Rialet Pieters Sekena H.Abdel-Aziem Anna M.vander Walt Cornelius C.Bezuidenhout John P.Giesy Mosaad A.Abdel-Wahhab Food Toxicology and Contaminants Department National Research CenterDokkiCairo 12622Egypt Unit of Environmental Sciences and Management North-West UniversityPrivate Bag X6001Potchefstroom 2520South Africa Cell Biology Department National Research CenterDokkiCairo 12622Egypt Department of Veterinary Biomedical Sciences and Toxicology Centre University of SaskatchewanSaskatoonSaskatchewan S7N 5B4Canada Department of Zoology and Center for Integrative ToxicologyMichigan State UniversityEast LansingMI 48824USA Department of Biology and Chemistry and State Key Laboratory in Marine Pollution City University of Hong KongKowloonHong KongChina School of Biological Sciences University of Hong KongHong KongChina State Key Laboratory of Pollution Control and Resource Reuse School of the EnvironmentNanjing UniversityNanjing 210093JiangsuChina Department of Biology Hong Kong Baptist UniversityHong KongChina

Aim:Protective effects of aqueous extract of Amaranthus hybridus against afl atoxin B1(AFB_(1))and/or fumonisin B1(FB_(1))on the H4IIE-luc cell line were determined by use of the methyl thiazol tetrazolium viability assay and disruption of DNA integrity.Methods:H4IIE-luc cells were incubated with different concentrations of AFB_(1) and/or FB_(1) for 24 and 48 h with or without aqueous extract of A.hybridus.Results:AFB_(1) decreased the viability of cells after 24 and 48 h of exposure.EC_(50)values for AFB_(1) were 10.5 and 1.8μmol/L for the two periods,respectively.When the 48 h exposure to mycotoxin repeated with a pre-treatment of 20 and 40μg/mL extract of A.hybridus,the EC_(50)changed to 3.88 and 7.67μmol/L,respectively.H4IIE-luc cells exposed to FB_(1) for 24 h responded more than those incubated for 48 h.cells treated with a combination of AFB_(1) and FB_(1) were less viable with a signifi cant decrease in the greater concentration.The mixture of AFB_(1) and FB_(1) resulted in a signifi cant threat to H4IIE-luc as indicated by the absence or appearance of new bands in random amplifi ed polymorphic DNA analysis,which demonstrated damage to DNA.The protective effects were probably due to greater content of total phenolics,carotenoids,β-carotene,folic-,linolenic-,linoleic and palmitic acids,as well as calcium,magnesium,iron,zinc,and selenium observed in the extract.Conclusion:Exposure to 40μg/mL of extract of A.hybridus protected cells from damage to DNA by stabilizing DNA.

关键词： Afl atoxin B1 Amaranthus hybridus cytotoxicity DNA fumonisin B1 hepatoma cells methyl thiazol tetrazolium assay

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Antisense EGFR sequence enhances apoptosis in a human hepatoma cell line BEL-7404

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Cell Research 1996年第2期6卷 145-153页

作者： FU TAO HE LIU FAN LIU JUN GU WAN LI JIANG YONG HUA XU (Laboratory of Cellular and Molecular Oncology, Shanghai Institute of Cell Biology, Chinese Academy of Sciences,Shanghai 200031)(Shanghai Bureau of Hygiene)(Laboratory of Molecular Biology, Nav Laboratory of Cellular and Molecular Oncology Shanghai Institute of Cell Biology Chinese Academy of Sciences Shanghai 200031 Shanghai Bureau of Hygiene Laboratory of Molecular Biology Naval Medical Research Institute Shanghai 200433 China

Effects of antisense epidermal growth factor receptor (EGFR) sequence on apoptotic cell death were examined in a human hepatoma cell line BEL-7404 cells. In the cells of JX-1, a sub clone of BEL-7404 stably transfected with antisense EGFR vector (Cell Research, 3:75, 1993), an enhanced rate (9.5%) of spontaneous apoptosis was detected by flow cytometry, whereas the rates of spontaneous apoptosis in JX-0 cells, a sub-clone of BEL-7404 transfected by control vector, and the parellt BEL-7404 cells were almost equal and about 1.7%. Serum-starvation for 72 h increased the rate of apoptosis of JX-1cells up to 33.7%, while JX-0 and BEL-7404 cells, under the same condition, produced less than 5% of apoptotic cells. Observation with electron microscope demonstrated that condensation and fragmentation of chromatin and formation of apoptotic bodies of ten occurred in JX-1 cells, especially during serumstarvation. These results, combined with the data of DNA fragmentation Elisa test, suggested that antisense EGFR sequence enhances apoptosis in the human hepatoma cells.Comparison of intracellular Ca2+ level and the responsiveness of JX-1 cells to the induced action of EGF and tharpsigargin (TG) treatment with that of control JX-0cells indicated that antisense EGFR might interrupt the EGF/EGFR signaling pathway resulting in the decreass of intracellular Ca2+ pool content as well as the responsiveness of these cells to the extracellular signals. These findings suggest that antisense EGFR either directly or indirectly reglllates Ca2+ storage in endoplasmic reticulum,thereby enhances apoptosis in the hnman hepatoma cells.

关键词： Antisense EGFR hepatoma cells apoptosis

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The selected killig of hepatocellular carcinoma cells using HSV/TK with The PBI-SUR-TK Vector

The selected killig of hepatocellular carcinoma cells using ...

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第六届泛环渤海生物化学与分子生物学会学术交流会

作者： Meng Fan-xiu Zhang Qi Yuan Yang-yang Yao Zhi-jian Zeng Si-heng Zhang Yue-hong Guo Rui Wang Hai-long Xie Jun Yu Bao-feng Department of Biochemistry and molecular biology Shanxi Medical University

The present study was performed to investigate the selective cytotoxicity of PBI-SUR-TK suicide gene expression level in HuH-7 hepatoma cells driven by the Survivin promoter.To evaluate thecytotoxicity of PBI-SUR-TK suicide gene on hepatoma cells.It can lay a foundation for the liver cancer gene therapy.Methods:The plasmid was transfected into the HuH-7 hepatoma cells with liposomes,we detected the expression levels of PBI-SUR-TK suicide gene in normal liver cells and hepatoma cells by RT-PCR and Western-blotting method.In addition,transfecting PBI-SUR-TK plasmid vector into HuH-7 hepatoma cells with lipoplexes vectors,with increasing of ganciclovir doses we explored the cytotoxicity of PBI-SUR-TK plasmid vector on hepatoma cells with CCK-8 reagents and Flow Cytometry.Results:The PBI-SUR-TK plasmid vector expression level was higher in hepatoma cells than in the control group without transfecting the plasmid.Additionally,CCK-8 assay indicated,increasing the dose of ganciclovir into the cell with transfected the plasmid vector,the cytotoxicity of PBI-SUR-TK suicide gene gradually increased.With Flow Cytometry,the cytotoxicity of the transfected hepatoma cells with the vector and untransfected cells were(36.43±4.02)% and(1.0±0.62)% separately with the difference being significant among them(Phepatoma cells with different doses of GCV.What’ s more,Our experiments laid a foundation for the liver-targeted gene therapy.

关键词： hepatoma cells targeted gene therapy HSV/TK

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