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STUDY OF ENHANCED IMMUNOGENECITY OF B7-1 GENE TRANSFECTED HUMAN hela cell line

Chinese Journal of Cancer Research 1998年第1期10卷 8-10页

作者：何曦秦慧莲向荣张跃建叶闻斐何球藻 Department of Immunology Shanghai Medical University

This work was supposed by CMB (No. 96—635) This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 1997). Although cervical carcinoma cells may express the human papillomavirus protein E6 and E7, they fail to induce an effective specific cytotoxic T lymphocyte response. Recent studies suggest that expression of CD 80 (B7 1) on tumor cells is effective to induce antitumor immune responses. 1,2 In our study, CD 80 gene was transfected into human hela cell line with a CD 80 expression plasmid (B7 1 +pcDNA 3) by electroporation, then the immunogenecity of the modified hela cell was tested in TLMC (tumor lymphocyte mixed culture) system. Thymidine lymphocyte proliferation assays showed that the response of human peripheral blood lymphocytes (PBLS) to CD 80 positive hela cells demonstrated a substantial increase in cell proliferation compared to the response to control cells. Cocultivation of allogeneic PBLs with CD 80 positive tumor cells for three days can induce an increased secretion of IL 2. Our results demonstrate an immunostimulatory effect of CD 80 expression on cervical cancer cells, which provides a basis for the development of a therapeutic tumor vaccine.

关键词： B7 1 gene hela cell line CD 80 Immuno genecity

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Withania somnifera Root Extract Enhances Telomerase Activity in the Human hela cell line

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Advances in Bioscience and Biotechnology 2016年第4期7卷 199-204页

作者： Vasantharaja Raguraman Jamuna R. Subramaniam Center for Preclinical and Translational Medicine Research Central Research Facility Sri Ramachandra University Chennai India

Aging is a decelerating unidirectional process of life. Shortening of telomeric DNA, the (TTAGGG)n hexanucleotide repeats, which form the caps at the chromosome ends, is implicated to determine the aging process, and more importantly the healthy lifespan itself. Telomerase, a ribonucleoprotein having reverse transcriptase activity, arrests telomere loss through addition of the TTAGGG repeats de novo, to the ends of the chromosome. The telomere/telomerase maintenance is an inevitable necessity to delay aging and for a healthy lifespan. Here, we report the potential of full-spectrum, high concentration Ashwagandha (Withania somnifera), an Ayurvedic medicinal herb, root extract to increase telomerase activity. hela cells, when treated with various concentrations of Ashwagandha root extract, showed an increase in telomerase activity measured with the established Telomerase Rapid Amplification Protocol (TRAP) assay. Ashwagandha root extract increased telomerase activity with highest enhancement of ~45% at 10 - 50 μg concentration. Thus, Ashwagandha root extract has the anti-aging inducing potential.

关键词： Ashwagandha Telomerase hela cell line TRAP Assay

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Microcalorimetric Study of Energy Metabolism of HSV-2 and FMDV Infection Processes

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Chinese Journal of Chemistry 2006年第2期24卷 182-186页

作者：张恒郑从义李杰顾朝江汪存信刘欲文 College of Chemistry and Molecular Science Wuhan University Wuhan Hubei 430072 China College of Life Science Wuhan University Wuhan Hubei 430072 China

The metabolic thermogenic power data of the HSV-2 infected hela cells and the FMDV infected BHK-21 cells were determined by LKB-2277 bioactivity monitor. The aim of the study was to investigate the difference of the cell metabolism under the action of two different viruses and the effects of hyperthermia and drugs on it. The results illustrated that the metabolic thermogenic power of infected cells was larger than the uninfected ones and there was a significant difference between the metabolism heat released by the two types of infected cells. From the maximal thermal power and total metabolism heat, the infection process was observed to be thermosensitive and could be inhibited by interferon. Our experiments also revealed that 6 month storage of FMDV could attenuate its virulence and infectivity. The study shows that microcalorimetry is a potent tool to investigate the metabolism of virus infection process.

关键词： HSV-2 FMDV hela cell line BHK-21 cell line metabolism microcalorimetry

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A DNA-based nanocarrier for efficient cancer therapy

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Journal of Pharmaceutical Analysis 2021年第3期11卷 330-339页

作者： Muhammad Abbas Mirza Muhammad Faran Ashraf Baig Yaliang Zhang Yu-Shun Yang Songyu Wu Yiqiao Hu Zhong-Chang Wang Hai-Liang Zhu State Key Laboratory of Pharmaceutical Biotechnology Nanjing UniversityNanjing210023PR China State Key Laboratory of Analytical Chemistry for Life Sciences School of Chemistry and Chemical EngineeringNanjing UniversityNanjing210023PR China Institute of Drug Research and Development Medical School of Nanjing UniversityNanjing210093PR China

The study aimed to achieve enhanced targeted cytotoxicity and cell-internalization of cisplatin-loaded deoxyribonucleic acid-nanothread(CPT-DNA-NT),mediated by scavenger receptors into hela ***-NT was developed with stiff-topology utilizing circular-scaffold to encapsulate *** force microscopy(AFM)characterization of the DNA-NT showed uniformity in the structure with a diameter of 50-150 nm and length of 300-600 *** successful fabrication of the DNA-NT was confirmed through native-polyacrylamide gel electrophoresis analysis,as large the molecular-weight(polymeric)DNA-NT did not split into constituting strands under applied current and *** results of cell viability confirmed that blank DNA-NT had the least cytotoxicity at the highest concentration(512 nM)with a viability of 92%as evidence of its biocompatibility for drug *** assay showed superior cytotoxicity of CPT-DNA-NT than that of the free CPT due to the depot release of CPT after DNA-NT *** DNA-NT exhibited targeted cell internalizations with the controlled intracellular release of CPT(from DNA-NT),as illustrated in confocal ***,in vitro cytotoxicity assessment through flow cytometry showed enhanced apoptosis(72.7%)with CPT-DNA-NT(compared to free CPT;64.4%).CPT-DNA-NT,being poly-anionic,showed enhanced endocytosis via scavenger receptors.

关键词： DNA-nanothread Cisplatin Drug delivery Scavenger receptors Caveolae hela cell line

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Comparative antitumor and anti-proliferative activities of Hippophae rhamnoides L. leaves extracts

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Journal of Coastal Life Medicine 2015年第3期3卷 228-232页

作者： Javid Ali Bashir Ahmad PCSIR Laboratories Complex Jamrude Road PeshawarKPK-Pakistan Center of Biotechnology and Microbiology University of PeshawarKPK-Pakistan

Objective:To evaluate the antitumor and anti-proliferative activities of methanol,aqueous,acetone,ethyl acetate,ethanol,chloroform and n-hexane extracts of Hippophae rhamnoides ***:Antitumor activities were evaluated by using the antitumor potato disc assay by using inoculums(Agrobacterium tumefaciens)with three different concentrations of test samples(10,100 and 1000 mg/L).Anti-proliferative activity was evaluated by the given method of methyl thiazolyl tetrazolium *** concentrations of the extract ranging from 0.039 to 10 mg/mL were tested against hela ***:Highest tumors inhibition activity(60.9%and 55.8%)was shown by methanol and ethanol extracts,with EC_(50) values of 424.41 and 434.61 mg/L *** 10 mg/mL,The highest cell inhibition 75.61%was observed in methanol extract and the lowest 36.59%were calculated in n-hexane *** difference in tumor and cell inhibition(%)may be due to the different concentration of active compounds responsible for antitumor and anti-proliferative *** extracts have considerable level of tumor and cell inhibitiory effect in a dose dependent ***:Our finding showed that Hippophae rhamnoides leaves are a potent natural source of antitumor and antiproliferative agent.

关键词： Seabuckthorn Solvent extracts Tumor inhibition Anti-cancer Potato disc assay hela cell line

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In Vitro Biological Activity of Anti-C Ⅱ TA Hammerhead Ribozyme——A Novel Approach for Autoimmune Diseases

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第4期23卷 335-338页

作者：刘芳邹萍郭荣陆华中范华骅 Institute of Hematology Department of Hematology Department of Blood Engineering

This study investigated the feasibility of using an hammerhead ribozyme against C Ⅱ TA, a major regulator of MHC Ⅱ antigens, to repress the expression of MHC Ⅱ molecules on hela cells. A hammerhead ribozyme (Rz464) specific to 463-465 GUC triplet of C Ⅱ TA and its target gene were transcribed, then mixed up and incubated in vitro . The cleavage products were analyzed by PAGE and silver staining. Rz464 was then inserted into the pIRES2 EGFP vector (pRz464). Stable transfectants of hela with pRz464 were tested for class Ⅱ MHC induction by recombinant human interferon gamma (IFN γ). mRNA of C Ⅱ TA was measured by RT PCR. Our results showed that Rz464 could exclusively cleave C Ⅱ TA RNA. When induced with IFN γ, the expression of HLA DR, DP, DQ on pRz464 + hela was induced, and the mRNA content of C Ⅱ TA decreased too. It is concluded that Rz464 could inhibit C Ⅱ TA and thus the family of genes was regulated by C Ⅱ TA:MHC Ⅱ molecules. These results provided insight into the future application of Rz464 as a new nucleic acid drug against auto immune diseases.

关键词： MHC class Ⅱ transactivator (C Ⅱ TA) hammerhead ribozyme hela cell line autoimmune disease

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The effects and mechanisms of cytoplasmic Macrophage colony-stimulating factor (M-CSF) on the proliferation,migration and invasion of hela cells

The effects and mechanisms of cytoplasmic Macrophage colony-...

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两岸三地药理学与临床药理学术会议

作者： ZHANG Meng-xia~2 WU Hai-yan~1 TU Jian~1 ZHANG Xiao-hong~2 LE Xiao-yong~1 TANG Sheng-song~(1,2) (1. Institute of Pharmacy and Pharmacology, University of South China, Hengyang 421001, China 2. Center for Life Sciences, University of South China, Hengyang 421001, China)

Objective To explore the effects and mechanisms of cytoplasmic M-CSF on the proliferation,mi- gration and invasion of hela *** Both pCMV/cyto/myc vector and pCMV/cyto/myc-M-CSF vector was transfected into hela-cell by *** screening by G418,the positive clones were amplified and confirmed by RT-PCR,Western blot and *** effect of cytoplasmic M- CSF on the proliferation of hela cells were analyzed by cell conuting and antisense *** migration and invasion of cell was measured by in vitro Transwell assay and Matrigel-coated polycarbonate *** expression of cycline,cyclinD1/2/3,CDK2/4/6,Rac1,and matrix metalloproteinase 2 and 9 (MMP2/9)were assayed by semiquantitative *** expression of bothα-tubulin and cdc42 were displayed by *** activity of MMP2 was detected by gelatin *** Re- suits A cell line(referred as to hela-M cell)that highly expresses cytoplasmic M-CSF was successfully es- tablished in the *** result indicated that hela-M cell had a larger volume,faster growth rate and shorter doubling time than either pCMV/cyto/myc transfected hela cells(referred as to hela-C cell)or untransfected hela cells(referred as to hela cell).M-CSF-specific antisense oligonucleoside significantly inhibited hela-M cell proliferation and had little effect on either hela-C cell or HeI.a-C cell ***- plasmic M-CSF up-regulated both the expression of cycline,cyclinD1 and cyclinD3,CDK2,CDK 4 and CDK6,a Rho GTPase ralative protein(Racl),cdc42 and MMP2,but had little effect on expression of MMP9 and cyclin ***,cytoplasmic M-CSF induced the rearrangement of the a-tubulin in hela cells and significantly promoted the migration and invasion of hela cells in *** Cyto- plasmic M-CSFs up-regulate the expression of cycline,cyclinD1 and cyclinD3,CDK2,CDK 4 and CDK6 and induces the proliferation of hela *** M-CSFs up-regulate the expression of

关键词： M-CSF hela cell line cytoskeleton migration invasion

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