scholarly journals PLANTLET REGENERATION FROM ACIFLUORFEN-TOLERANT CELL LINES OF SOLANUM PTYCANTHUM AND LYCOPERSICON PERUVIANUM.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1082b-1082
Author(s):  
Chang-Yeon Yu ◽  
John Masiunas
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cultured Cells ◽  
Lycopersicon Peruvianum ◽  
Shoot Height ◽  
Solanum Ptycanthum ◽  
Friable Callus ◽  
Wide Range ◽  
Selection Medium ◽  
Tolerant Cell

Friable callus of Solanum ptycanthum and L. peruvianum PI199380 clone 149 were subcultured on liquid Murashige and Skoog salts and Gamborg Vitamin medium with 2,4-D (1mg/l) until a fine suspension of cells was obtained. The suspension cultured cells were then plated on selection medium. Twenty-five acifluorfen-tolerant cell lines of Solanum ptycanthum and fourteen tolerant Lycopersicon peruvianum cell lines were obtained by a stepwise increase in concentration of acifluorfen. Acifluorfen-tolerant cell lines were transferred on to regeneration media with the herbicide. Shoot regeneration differed depending on the cell line and acifluorfen concentration, ranging from 0 to 37 plants per calli. As acifluorfen concentration increased in the regeneration media, the number of shoots and shoot height decreased. There was a wide range of variation in shoot morphology, which depended on the cell line.

scholarly journals Recombinant deoxyribonucleoside kinase from Drosophila melanogaster can improve gemcitabine based combined gene/chemotherapy for targeting cancer cells

2019 ◽  
Author(s):  
Mahak Fatima ◽  
M. Mubasshar Iqbal Ahmed ◽  
Faiza Batool ◽  
Anjum Riaz ◽  
Moazzam Ali ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Cancer Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Nucleoside Analogs ◽  
Cancer Cell Lines ◽  
Control Group ◽  
Wide Range

A recombinant deoxyribonucleoside kinase from Drosophila melanogaster with a deletion of the last 20 amino acid residues (named DmdNKΔC20) was hypothesized as a potential therapeutic tool for gene therapy due to its broad substrate specificity and better catalytic efficiency towards nucleosides and nucleoside analogs. This study was designed to evaluate the effect of DmdNKΔC20 for sensitizing human cancer cell lines towards gemcitabine and to further investigate its role in reversal of acquired drug resistance in gemcitabine-resistant cancer cell line. The DmdNKΔC20 gene was delivered to three different cancer cell lines, including breast, colon and liver cancer cells, using lipid-mediated transfection reagent. After transfection, gene expression of DmdNKΔC20 was confirmed by reverse transcription quantitative PCR (qRT-PCR) and the combined effect of DmdNKΔC20 and gemcitabine based cytotoxicity was observed by cell viability assay. We further evolved a gemcitabine-resistant breast cancer cell line (named MCF7-R) through directed evolution in the laboratory, which showed 375-fold more resistance compared to parental MCF7 cells. Upon transfection with DmdNKΔC20 gene, MCF7-R cells showed 83-fold higher sensitivity to gemcitabine compared to the control group of MCF7-R cells. Moreover, we observed 79% higher expression of p21 protein in transfected MCF7-R cells, which may indicate induction of apoptosis. Our findings highlight the importance and therapeutic potential of DmdNKΔC20 in combined gene/chemotherapy approach to target a wide range of cancers, particularly gemcitabine-resistant cancers.

Biological Properties of Cell Lines Derived from Moloney Virus-Induced Sarcoma

1976 ◽  
Vol 62 (4) ◽  
pp. 415-428 ◽  
Author(s):  
Aurelio Di Marco ◽  
Teresa Dasdia ◽  
Fernando Giuliani ◽  
Anna Necco ◽  
Anna Maria Casazza ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Chromosome Number ◽  
Cell Line ◽  
Cell Lines ◽  
Acrocentric Chromosome ◽  
Biological Properties ◽  
Modal Number ◽  
Virus Content ◽  
Wide Range

Two cell lines derived from a primary MSV-M-induced tumor in a BALB/c mouse were studied. One line (MS-2) was subject only to continuous tissue culture transfer (tct). After 21 tct, MS-2 cells produced progressive tumors (MS-2 tumors) in syngeneic hosts. The second cell line (MS-2T) was established by cultivation of a MS-2 tumor. The ability to produce progressive tumors decreased with increased number of tct, in both cell lines. The virus content of MS-2 and MS-2T cells was very low, as shown by uridine incorporation and electron microscopy. Immunofluorescence tests demonstrated that antigens different from the viral MSV-M antigens were present on the cell lines, and that antigenic changes occurred with increased number of tct. Serum of mice bearing progressive MS-2 tumors reacted with MS-2T cells when these cells produced progressive tumors and did not react with MS-2 cells when they produced regressing tumors. MS-2 cells producing regressing tumors reacted with serum from mice in which the MS-2 tumor had regressed and with serum from mice immunized with MS-2T cells at late tct when they were poorly oncogenic. The antigenic changes seemed, therefore, to parallel the decrease of malignancy. A chromosomal analysis carried out on MS-2 and MS-2T cells, when both produced progressive tumors, showed a modal number of 48 and 44, respectively. MS-2T cells showed a large acrocentric chromosome. In contrast, the MS-2 cells at late tct, when they gave regressing tumors, showed a modal number of 60 and a wide range of distribution of chromosome number.

scholarly journals Cflar Splicing SNP As Predictor of Chemo-Sensitivity to Anticancer Drugs

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2056-2056
Author(s):  
Lata Chauhan ◽  
Emilie J Bergsma ◽  
Jatinder K Lamba
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Anticancer Drugs ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Chemotherapeutic Agents ◽  
Extrinsic Pathway ◽  
Wide Range

Abstract Background: Anticancer therapeutics leverages activation of apoptosis signal transduction pathways (extrinsic and intrinsic apoptotic pathways) in cancer cells. Apoptosis induced by the extrinsic pathway complements that induced by the intrinsic pathway, so targeting extrinsic pathway is considered a useful new therapeutic approach. Preclinical data suggests TNF related apoptosis inducing ligand (TRAIL) as a promising approach as apoptosis of tumor cells is achievable in vivo without lethal toxicities. CASP8 and FADD-like apoptosis regulator (CFLAR) is an inhibitor of death receptor signaling that inhibits TRAIL-mediated caspase 8 auto-activation and subsequent apoptosis. We recently identified a splicing single nucleotide polymorphism (SNP) rs10190751 G>A in CFLAR, where presence of the variant allele (A) was associated with alternate splicing as well as with chemo-sensitivity to chemotherapeutic agent triptolide. However role of CFLAR and the splicing SNP on chemo-sensitivity to wide array of anticancer drugs is not known. Objective: Given the central role of CFLAR in apoptotic pathway, the goal of this study was to investigate impact of CFLAR and its splicing SNP on cytotoxicity of wide range of chemotherapeutic drugs including the ones extensively used in hematological malignancies. Methods: We selected chemotherapeutic agents with wide range of mechanisms of action as blocking DNA biosynthesis, interfering with structure or function of DNA or protein synthesis, interfering with DNA transcription or replication as well as drugs that are cell cycle specific or not. We selected nine Epstein-Barr-virus transformed lymphoblastoid cell lines (LCLs) that are part of International HapMap project representing different genotype for rs10190751 (CFLAR splicing polymorphism; 3 in each genotype category) with twelve different chemotherapeutic agents. Further validation of CFLAR's role in in vitro chemosensitivity was evaluated using CFLAR knockdown and overexpression studies in pancreatic and leukemic cell lines such as Panc-1 and THP1. Results: CFLAR splicing SNP rs10190751, was associated with in vitro cytotoxicity of several chemotherapeutic agents (Bortezomib, SAHA, doxorubicin, sorafenib). The results of screening of 122 FDA approved drugs and their relation with CFLAR as well as its splicing SNP will be presented at the annual meeting. As an example we show below that knock down of CFLAR isoforms have a significant impact on in vitro chemosensitivity to bortezomib and SAHA (Figure 1) Conclusion: Our results suggest critical role of CFLAR in anticancer drug mediated cell death. Additionally splicing SNP in CFLAR seems to play an important role in drug sensitivity/resistance. Therapeutic strategies to directly or indirectly inhibit the expression and/or function of CFLAR might be an attractive option to overcome resistance to wide range of chemotherapeutic agents. Figure 1. Impact of siRNA mediated knockdown or of CFLAR on Bortezomib and SAHA sensitivity in THP1 and Panc-1 cancer cell line. Figure 1. Impact of siRNA mediated knockdown or of CFLAR on Bortezomib and SAHA sensitivity in THP1 and Panc-1 cancer cell line. Disclosures No relevant conflicts of interest to declare.

scholarly journals Exploration of Benzenesulfonamide-Bearing Imidazole Derivatives Activity in Triple-Negative Breast Cancer and Melanoma 2D and 3D Cell Cultures

2021 ◽  
Vol 14 (11) ◽  
pp. 1158
Author(s):  
Benas Balandis ◽  
Vytautas Mickevičius ◽  
Vilma Petrikaitė
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Triple Negative Breast Cancer ◽  
Heterocyclic Compounds ◽  
Triple Negative ◽  
Biological Activities ◽  
Human Malignant Melanoma ◽  
Imidazole Derivatives ◽  
Wide Range

Heterocyclic compounds are one of the main groups of organic compounds possessing wide range of applications in various areas of science and their derivatives are present in many bioactive structures. They display a wide variety of biological activities. Recently, more and more attention has been focused to such heterocyclic compounds as azoles. In this work, we have synthesized a series of new imidazole derivatives incorporating a benzenesulfonamide moiety in their structure, which then were evaluated for their cytotoxicity against human triple-negative breast cancer MDA-MB-231 and human malignant melanoma IGR39 cell lines by MTT assay. Benzenesulfonamide-bearing imidazole derivatives containing 4-chloro and 3,4-dichlorosubstituents in benzene ring, and 2-ethylthio and 3-ethyl groups in imidazole ring have been determined as the most active compounds. Half-maximal effective concentration (EC50) of the most cytotoxic compound was 27.8 ± 2.8 µM against IGR39 cell line and 20.5 ± 3.6 µM against MDA-MB-231 cell line. Compounds reduced cell colony formation of both cell lines and inhibited the growth and viability of IGR39 cell spheroids more efficiently compared to triple-negative breast cancer spheroids.

scholarly journals Human argininosuccinate synthetase minigenes are subject to arginine-mediated repression but not to trans induction.

1986 ◽  
Vol 6 (4) ◽  
pp. 1244-1252 ◽  
Author(s):  
F M Boyce ◽  
G M Anderson ◽  
C D Rusk ◽  
S O Freytag
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Transcription Initiation ◽  
Cultured Cells ◽  
Argininosuccinate Synthetase ◽  
Flanking Sequence ◽  
Cell Type Specificity ◽  
Resistant Variant ◽  
Acting Mechanism ◽  
Cat Expression

The human argininosuccinate synthetase locus is subject to metabolite-mediated repression by arginine in some cultured cell lines. To gain insight into the mechanism underlying this regulation, chloramphenicol acetyltransferase (CAT) minigenes under the transcriptional control of the human argininosuccinate synthetase promoter were constructed and tested for regulation. When the minigenes were introduced into RPMI 2650 cells, a human cell line that shows sixfold regulation of the argininosuccinate synthetase gene, CAT expression was repressed three- to fivefold when arginine was present in the culture medium. A minigene containing only 149 base pairs of 5'-flanking sequence was expressed at similar levels and regulated to the same degree as one having approximately 3 kilobases of 5'-flanking sequence. Therefore, the cis-acting sequences required for the arginine-mediated repression are likely to be located within the region of the transcription initiation site. The arginine-mediated repression of the CAT minigenes was not observed in canavanine-resistant variants of RPMI 2650 cells, and therefore they showed the appropriate cell-type specificity. Cultured cells having 200-fold-increased levels of argininosuccinate synthetase can be selected by growth in medium containing the arginine analog canavanine. It was previously demonstrated that the increased expression of argininosuccinate synthetase in canavanine-resistant human lymphoblasts was due to a trans-acting mechanism. To gain further support for a trans-acting mechanism, we tested our CAT minigenes for the trans induction in canavanine-resistant variants of RPMI 2650 cells. Transfection of the CAT minigenes into RPMI 2650 cells and canavanine-resistant variants of this cell line yielded no difference in transient CAT expression. Furthermore, cloned canavanine-resistant variant cells having integrated copies of the CAT minigenes expressed CAT at similar levels as compared to the parental cell lines. Since these cell lines do exhibit arginine-mediated repression of CAT but not trans induction, these data indicate that the argine-mediated repression is a regulatory event that occurs independently of the trans induction.

scholarly journals Evidence for the Need to Evaluate More Than One Source of Extracellular Vesicles, Rather Than Single or Pooled Samples Only, When Comparing Extracellular Vesicles Separation Methods

Cancers ◽  
2021 ◽  
Vol 13 (16) ◽  
pp. 4021
Author(s):  
Sarai Martinez-Pacheco ◽  
Lorraine O’Driscoll
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Extracellular Vesicles ◽  
Cultured Cells ◽  
Cancer Cell Lines ◽  
Breast Cancer Cell Lines ◽  
Pooled Samples

To study and exploit extracellular vesicles (EVs) for clinical benefit as biomarkers, therapeutics, or drug delivery vehicles in diseases such as cancer, typically we need to separate them from the biofluid into which they have been released by their cells of origin. For cultured cells, this fluid is conditioned medium (CM). Previous studies comparing EV separation approaches have typically focused on CM from one cell line or pooled samples of other biofluids. We hypothesize that this is inadequate and that extrapolating from a single source of EVs may not be informative. Thus, in our study of methods not previous compared (i.e., the original differential ultracentrifugation (dUC) method and a PEG followed by ultracentrifugation (PEG + UC) method), we analyzed CM from three different HER2-positive breast cancer cell lines (SKBR3, EFM192A, HCC1954) that grow in the same culture medium type. CM from each was collected and equally divided between both protocols. The resulting isolates were compared on seven characteristics/parameters including particle size, concentration, structure/morphology, protein content, purity, detection of five EV markers, and presence of HER2. Both dUC and PEG + UC generated reproducible data for any given breast cancer cell lines’ CM. However, the seven characteristics of the EV isolates were cell line- and method-dependent. This suggests the need to include more than one EV source, rather than a single or pooled sample, when selecting an EV separation method to be advanced for either research or clinical purposes.

scholarly journals The Curious Case of the HepG2 Cell Line: 40 Years of Expertise

2021 ◽  
Vol 22 (23) ◽  
pp. 13135
Author(s):  
Viktoriia A. Arzumanian ◽  
Olga I. Kiseleva ◽  
Ekaterina V. Poverennaya
Keyword(s):  
Liver Cancer ◽  
Cell Line ◽  
Hepg2 Cell ◽  
Cell Lines ◽  
Molecular Mechanisms ◽  
Basic Research ◽  
Health Concern ◽  
Hepg2 Cell Line ◽  
Wide Range

Liver cancer is the third leading cause of cancer death worldwide. Representing such a dramatic impact on our lives, liver cancer is a significant public health concern. Sustainable and reliable methods for preventing and treating liver cancer require fundamental research on its molecular mechanisms. Cell lines are treated as in vitro equivalents of tumor tissues, making them a must-have for basic research on the nature of cancer. According to recent discoveries, certified cell lines retain most genetic properties of the original tumor and mimic its microenvironment. On the other hand, modern technologies allowing the deepest level of detail in omics landscapes have shown significant differences even between samples of the same cell line due to cross- and mycoplasma infection. This and other observations suggest that, in some cases, cell cultures are not suitable as cancer models, with limited predictive value for the effectiveness of new treatments. HepG2 is a popular hepatic cell line. It is used in a wide range of studies, from the oncogenesis to the cytotoxicity of substances on the liver. In this regard, we set out to collect up-to-date information on the HepG2 cell line to assess whether the level of heterogeneity of the cell line allows in vitro biomedical studies as a model with guaranteed production and quality.

scholarly journals Isolation in Natural Host Cell Lines of Wolbachia Strains wPip from the Mosquito Culex pipiens and wPap from the Sand Fly Phlebotomus papatasi

Insects ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 871
Author(s):  
Lesley Bell-Sakyi ◽  
Alexandra Beliavskaia ◽  
Catherine S. Hartley ◽  
Laura Jones ◽  
Lisa Luu ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Culex Pipiens ◽  
Host Cells ◽  
Sand Fly ◽  
Phlebotomus Papatasi ◽  
Wide Range ◽  
Tetracycline Treatment ◽  
Cell Line Generation ◽  
Line Generation

Endosymbiotic intracellular bacteria of the genus Wolbachia are harboured by many species of invertebrates. They display a wide range of developmental, metabolic and nutritional interactions with their hosts and may impact the transmission of arboviruses and protozoan parasites. Wolbachia have occasionally been isolated during insect cell line generation. Here, we report the isolation of two strains of Wolbachia, wPip and wPap, during cell line generation from their respective hosts, the mosquito Culex pipiens and the sand fly Phlebotomus papatasi. wPip was pathogenic for both new C. pipiens cell lines, CPE/LULS50 and CLP/LULS56, requiring tetracycline treatment to rescue the lines. In contrast, wPap was tolerated by the P. papatasi cell line PPL/LULS49, although tetracycline treatment was applied to generate a Wolbachia-free subline. Both Wolbachia strains were infective for a panel of heterologous insect and tick cell lines, including two novel lines generated from the sand fly Lutzomyia longipalpis, LLE/LULS45 and LLL/LULS52. In all cases, wPip was more pathogenic for the host cells than wPap. These newly isolated Wolbachia strains, and the novel mosquito and sand fly cell lines reported here, will add to the resources available for research on host–endosymbiont relationships, as well as on C. pipiens, P. papatasi, L. longipalpis and the pathogens that they transmit.

Epithelial organization and hormone sensitivity of toad urinary bladder cells in culture

1981 ◽  
Vol 241 (2) ◽  
pp. F129-F138 ◽  
Author(s):  
J. P. Johnson ◽  
R. E. Steele ◽  
F. M. Perkins ◽  
J. B. Wade ◽  
A. S. Preston ◽  
...  
Keyword(s):  
Urinary Bladder ◽  
Cell Line ◽  
Cell Lines ◽  
Water Permeability ◽  
Sodium Transport ◽  
Time Course ◽  
Cultured Cells ◽  
Transepithelial Transport ◽  
Seeding Density ◽  
Toad Urinary Bladder

Two continuous cell lines (TB-M and TB-6c) derived from epithelial cells of the toad urinary bladder form epithelia in culture that manifest hormone-sensitive transepithelial transport. Development of transepithelial electrical resistance (R) and transport rate (ISC) are dependent on time and density of cells seeded, but steady-state ISC and R are characteristic for each cell line and independent of seeding density. Some responses of intact toad bladder are preserved in culture, whereas others are altered or absent. Neither cell line responds to vasopressin. Analogues of cAMP increase sodium transport and urea permeability in both cell lines but do not affect water permeability. The intramembrane particle aggregates associated with the vasopressin- and cAMP-induced increase in water permeability of the intact bladder could not be detected in the cell lines. Aldosterone increases sodium transport in both cell lines, and the time course and concentration dependence of the response to aldosterone are similar to those of the intact bladder. The relative effect of a series of steroids on ISC reveals corticosterone to be a more potent mineralocorticoid in cultured cells than in the intact bladder.

Synthesis and Anti-Proliferative Evaluations of New Heterocyclic Derivatives using 5,6,8,9-tetrahydropyrazolo[5,1-b]quinazolin-7(3H)-one Derivatives Derived from Cyclohexa-1,4-dione

2020 ◽  
Vol 20 ◽  
Author(s):  
Mahmoud A.A. Mahmoud ◽  
Meshari A. Alsharif ◽  
Rafat M. Mohareb
Keyword(s):  
Lung Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Mtt Assay ◽  
Cancer Cell Lines ◽  
Positive Control ◽  
Human Lung Cancer ◽  
Wide Range ◽  
Target Molecules

Background: Recentlty pyrazoloquinazoline derivatives acquired a special attention due to their wide range of pharmacological activities especially the therapeutic activities. Through the market it was found that many pharmacological drugs containing the quinazoline nucleus were known. Objective: We are aiming in this work to synthesize target molecules not only possess anti-tumor activities but also kinase inhibitors. The target molecules were obtained through the synthesis of a series of 5,6,8,9-tetrahydropyrazolo[5,1-b]quinazolin-7(3H)-one derivatives 4a-i using the multicomponent reactions of cyclohexan-1,4-dione (1), the 5-amino-4-(2-arylhydrazono)-4H-pyrazol-3-ol derivatives 2a-c the aromatic aldehydes 3a-c, respectively. The synthesized compounds were evaluated against c-Met kinase, PC-3 cell line and different kinds of cancer cell lines together with normal cell line, tyrosine kinases and Pim-1 kinase. Methods: Muticomponent reactions were adopted using compound 1 to get different 5,6,8,9- tetrahydropyrazolo[5,1-b]quinazolin-7(3H)-one derivatives which underwent further heterocyclization reactions. The c-Met kinase activity of all compounds was evaluated using Homogeneous TimeResolved Fluorescence (HTRF) assay taking foretinib as the positive control. . The anti-proliferative activity of all target compounds against the human prostatic cancer PC-3 cell line were measured using MTT assay using SGI-1776 as the reference drug. All the synthesized compounds were assessed the inhibitory activities against A549 (non-small cell lung cancer), H460 (human lung cancer), HT-29 (human colon cancer) and MKN-45 (human gastric cancer cancer) cancer cell lines together with foretinib as the positive control by a MTT assay. Results: Antiproliferative evaluations and c-Met kinase, Pim-1 kinse inhibitions were perform for the synthesized compounds where the varieties of substituent through the aryl ring and the thiophene moiety afforded compounds with high activities. Conclusion: The compounds with high antiprolifeative activity were tested toward c-Met and the results showed that compounds 4e, 4f, 4g, 4i, 6i, 6k, 6l, 8f, 8i, 10d, 10e, 10f, 10h, 12e, 12f, 12g, 12h, 12i, 14f, 14g, 14h and 14i were the most potent compounds. Further selection of compounds for the Fused Quinazoline Derivatives as Anticancer Agents Pim-1 kinase inhibition activity showed that compounds 4f, 6i, 6l, 8h, 8i,8g, 10d, 12i and 14f were the most active compounds to inhibit Pim-1.

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