CYTOTOXIC POTENTIAL OF L-ASPARAGINASE FROM BACILLUS SP. IN VITRO

This study reports the cytotoxic potential of L-Asparaginase isolated from Bacillus sp. Bacillus sp. was isolated from local soil/water samples and identified by rapid plate assay and further confirmed by phenotypic characterization. Extracellular L- Asparaginase was isolated from broth culture of Bacillus sp. and purified by ammonium sulfate precipitation, followed by dialysis and ion exchange and gel filtration chromatography techniques. The purified enzyme was used to study the in vitro cytotoxic potential. Varying concentrations (31.25, 62.5, 125, 250, 500 µg/ml) of purified L-Asparaginase was tested on MCF7, HeLa, HepG2 and 3T3L1cell lines by MTT assay. Curcumin was maintained as a positive control. The results revealed that the enzyme showed a significant cytotoxic activity and a dose dependent effect. Minimum inhibition of (19.44%) was observed at an enzyme concentration of 31.25 µg/ml and maximum inhibition (71.14%) was observed at 500ug/ml against MCF7 cell line. Minimum inhibition of (10.04%) was observed at an enzyme concentration 31.25 µg/ml and maximum inhibition (68.92%) was observed at 500 µg/ml against HeLa cell line. Minimum inhibition of (7.45%) was shown at an enzyme concentration of 31.25 µg/ml and maximum inhibition (68.28 %) was observed at 500 µg/ml against HepG2 cell line. Minimum inhibition of (4.4%) was shown by enzyme concentration 31.25 µg/ml and maximum inhibition (47.4%) was observed at 500 µg/ml against 3T3L1 cell line. Curcumin at 5µM concentration showed an inhibition of 48.13% against MCF-7 cells, 54.42% against HeLa cells, 64.94% against HepG2 cells and 44.5% against 3T3L1 cell lines.

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Evaluation of cytotoxic potential of L-asparaginase from Scopulariopsis brevicaulis on cell lines in vitro

Biomedicine ◽

10.51248/.v39i2.189 ◽

2020 ◽

Vol 39 (2) ◽

pp. 254-257

Author(s):

D’Souza Renita Maria ◽

Abraham Asha

Keyword(s):

Cell Line ◽

Gel Filtration ◽

Positive Control ◽

Enzyme Concentration ◽

Cytotoxic Potential ◽

3T3l1 Cell ◽

Maximum Inhibition ◽

Scopulariopsis Brevicaulis ◽

Mcf7 Cell Line

Introduction and Aim: This study reports the cytotoxic potential of L-Asparaginase isolated from the fungus Scopulariopsis brevicaulis. Materials and Methods: Extracellular L- Asparaginase was isolated from Scopulariopsis brevicaulis and purified by ammonium sulfate precipitation, followed by dialysis, ion exchange and gel filtration chromatography. Varying concentrations (31.25, 62.5, 125, 250, 500 µg/ml) of purified L-Asparaginase was tested on MCF7, HeLa, HepG2 and 3T3L1cell lines by MTT assay. Curcumin was maintained as a positive control. Results: Minimum inhibition of 23.57% was observed at an enzyme concentration of 31.25 µg/ml and maximum inhibition (66.41%) was observed at 500 µg/ml against MCF7 cell line. Minimum inhibition of 2.87% was observed at an enzyme concentration 31.25 µg/ml and maximum inhibition (58.49%) was observed at 500 µg/ml against HeLa cell line. Minimum inhibition of 4.58% was shown at an enzyme concentration of 31.25 µg/ml and maximum inhibition (46.14 %) was observed at 500 µg/ml against HepG2 cell line. Minimum inhibition of 1.4% was shown by enzyme concentration 31.25 µg/ml and maximum inhibition (50.9%) was observed at 500 µg/ml against 3T3L1 cell line. Conclusion: We report for the first time the cytotoxic potential of L-Asparaginase from Scopulariopsis brevicaulis.

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In Vitro Cytotoxic Potential of Plant Terpenes and Selected Medicinal Plants of Pakistan against NIH 3T3 Cell Line.

Annals of King Edward Medical University ◽

10.21649/akemu.v24is.2551 ◽

2018 ◽

Vol 24 (S) ◽

pp. 853-858

Author(s):

Shazia Kanwal Malik ◽

Maqsood Ahmed ◽

Farah Khan

Keyword(s):

Medicinal Plants ◽

Cell Line ◽

Acid Methyl Ester ◽

Hexadecanoic Acid ◽

Cytotoxic Potential ◽

Reference Drug ◽

Animal Trials ◽

Diphenyl Tetrazolium Bromide ◽

Nih 3T3

Background: Medicinal plants are used in anticancer therapies from ages. The very first time we have not only screened anticancer medicinal plants from Pakistan but also identified anticancer plant terpenes. This study will definitely pave the way towards anticancer drug development after animal trials. Materials and Methods | This study was based on ethanolic extraction of fruits/seed of 25 plants. The cytotoxic potential of extracts and plant terpenes was evaluated against mouse fibroblasts (NIH 3T3) cell line by using MTT (3- [4, 5-dimethylthiazole-2-yl]-2, 5-diphenyl-tetrazolium bromide) assay and compared with cyclohexamide. Results | Ethanolic extracts were nontoxic against NIH3T3 cell line. On the basis of the % inhibition plants were graded as B.lycium, 48% > T.bellerica, 47% > C.verum, 45% > Z.tenuior, 39% > H. adenophyllum, 38% > C. carandas, 31% > C.behen, 29% > P. juliflora, 28% & P.granatum, 28% W.somnifera, 27% > E.cheiri, 26% > S. potatorum, 24% & M.incana, 24% > D. peregrina, 21% & B.lanzan, 21% > L. maldivica, 20% > F. lyrata, 18% > F.arabica, 16% & R. indica, 16% > C.absus, 13% & C.paniculatus, 13%. > C. diurnum, 12% > J.mimosifolia, 11% > D.malabarica, 8% > C. speciosa, 3% as compared to the reference drug Cyclohexamide, 70%. Whereas the maximum % inhibition was exhibited by n-Hexadecanoic acid (88%), followed by Hexadecanoic acid, methyl ester (16%) and Methyl decanoate displayed 11%. C50 of n-Hexadecanoic acid and Cyclohexamide was 3.1±0.2 and 0.8±0.2 respectively. Conclusion| Plant extracts and terpenes was found to be inactive against 3T3 cell line.

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Comparative Cytotoxicity Rankings of Four Aminoglycoside Antibiotics in the Chang, SIRC and LLC-PK1 Cell Lines

Alternatives to Laboratory Animals ◽

10.1177/026119299001800128.1 ◽

1990 ◽

Vol 18 (1_part_1) ◽

pp. 283-290

Author(s):

Daniel A. Linseman ◽

Timothy J. Raczniak ◽

C. Sidney Aaron ◽

James A. Bacon

Keyword(s):

Proximal Tubule ◽

Cell Line ◽

Cell Lines ◽

Cytotoxic Potential ◽

Corneal Epithelial ◽

Increased Sensitivity ◽

Basal Cytotoxicity ◽

Chang Liver

A series of structurally related aminoglycosides — neomycin, gentamicin, amikacin, and streptomycin — were screened for cytotoxicity in three cell lines, Chang (liver), SIRC (corneal epithelial), and LLC-PK1 (kidney). The main objectives of this study were: firstly, to determine whether the proximal tubule origin of the LLC-PK1 cell line conferred increased sensitivity to this class of xenobiotic when compared to cell lines derived from organs other than the kidney; and secondly, to determine whether any of the cell lines would rank the in vitro cytotoxic potential of the compounds in an order consistent with their in vivo toxicities. LDH leakage and cell proliferative effects (CP) were the endpoints used to measure cytotoxicity. The proximal tubule derivation of the LLC-PK1 cell line did not appear to confer significantly increased sensitivity to any of the aminoglycosides tested using LDH release and cell proliferation as endpoints of cytotoxicity. The relative cytotoxicity rankings were as follows: Chang — gentamicin>neomycin>amikacin>streptomycin (LDH), neomycin∼gentamicin∼streptomycin >amikacin (CP); SIRC — neomycin∼gentamicin∼streptomycin>amikacin (LDH and CP); and LLC-PK1 — gentamicin∼streptomycin>neomycin>amikacin (LDH), and streptomycin >neomycin>gentamicin∼amikacin (CP). The results suggest that the Chang line provides a cytotoxicity ranking consistent with in vivo nephrotoxicity data. The SIRC line ranks amikacin the least cytotoxic, but fails to discriminate between the cytotoxicities of gentamicin, neomycin and streptomycin. The LLC-PK1 cell line ranks the compounds in an order which is inconsistent with in vivo results. The LLC-PK1 cell line appears to be the most sensitive to streptomycin, which is the only agent tested that is not accumulated in the kidney in vivo. The results may reflect basal cytotoxicity, since relatively non-specific endpoints were used. Perhaps the LLC-PK1 cell line would rank the cytotoxic potential of this class of compounds more accurately if parameters which are more renal-specific were measured as endpoints.

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Biodegradation of cypermethrin by Bacillus sp. in soil microcosm and in-vitro toxicity evaluation on human cell line

International Biodeterioration & Biodegradation ◽

10.1016/j.ibiod.2012.11.008 ◽

2013 ◽

Vol 77 ◽

pp. 39-44 ◽

Cited By ~ 23

Author(s):

Smita Sundaram ◽

Mihir Tanay Das ◽

Indu Shekhar Thakur

Keyword(s):

Cell Line ◽

Human Cell ◽

Human Cell Line ◽

Soil Microcosm ◽

In Vitro Toxicity ◽

Bacillus Sp ◽

Toxicity Evaluation

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New platinum(IV) complex with adamantylamine ligand as a promising anti-cancer drug: comparison of in vitro cytotoxic potential towards A2780/cisR cisplatin-resistant cell line within homologous series of platinum(IV) complexes

Anti-Cancer Drugs ◽

10.1097/01.cad.0000127147.57796.e5 ◽

2004 ◽

Vol 15 (5) ◽

pp. 537-543 ◽

Cited By ~ 26

Author(s):

Jaroslav Tur??nek ◽

Andrea Ka??n?? ◽

Dana Z??lusk?? ◽

Ji???? Ne??a ◽

Veronika Kvardov?? ◽

...

Keyword(s):

Cell Line ◽

Homologous Series ◽

Resistant Cell ◽

Resistant Cell Line ◽

Cancer Drug ◽

Cytotoxic Potential ◽

Anti Cancer

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PHENOTYPIC CHARACTERIZATION OF TWO NOVEL CELL LINE MODELS OF CASTRATION RESISTANT PROSTATE CANCER.

10.1101/2021.07.04.450352 ◽

2021 ◽

Author(s):

Michael C. Haffner ◽

Akshay Bhamidipati ◽

Harrison K. Tsai ◽

David M. Esopi ◽

Ajay M. Vaghasia ◽

...

Keyword(s):

Prostate Cancer ◽

Cell Line ◽

Cell Lines ◽

Protein Level ◽

Phenotypic Characterization ◽

Castration Resistant Prostate Cancer ◽

Castration Resistance ◽

Castration Resistant

BACKGROUND: Resistance to androgen deprivation therapies is a major driver of mortality in advanced prostate cancer. Therefore, there is a need to develop new pre-clinical models that allow the investigation of resistance mechanisms and the assessment of drugs for the treatment of castration resistant prostate cancer. METHODS: We generated two novel cell line models (LAPC4-CR and VCaP-CR) which were derived by passaging LAPC4 and VCaP cells in vivo and in vitro under castrate conditions. We performed detailed transcriptomic (RNA-seq) and proteomic analyses (SWATH-MS) to delineate expression differences between castration-sensitive and castration-resistant cell lines. Furthermore, we characterized the in vivo and in vitro growth characteristics of the novel cell line models. RESULTS: The two cell line derivatives LAPC4-CR and VCaP-CR showed castration resistant growth in vitro and in vivo which was only minimally inhibited by AR antagonists, enzalutamide and bicalutamide. High-dose androgen treatment resulted in significant growth arrest of VCaP-CR but not in LAPC4-CR cells. Both cell lines maintained AR expression, but exhibited distinct expression changes on the mRNA and protein level. Integrated analyses including data from LNCaP and the previously described castration resistant LNCaP-abl cells revealed an expression signature of castration resistance. CONCLUSIONS: The two novel cell line models LAPC4-CR and VCaP-CR and their comprehensive characterization on the RNA and protein level represent important resources to study the molecular mechanisms of castration resistance.

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Interferon gamma resistance in setting of LKB1 loss: Phenotypic characterization and investigation of mechanism.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.15_suppl.e21015 ◽

2020 ◽

Vol 38 (15_suppl) ◽

pp. e21015-e21015

Author(s):

Jacob Kaufman

Keyword(s):

T Cell ◽

Cell Line ◽

Cell Lines ◽

Interferon Gamma ◽

Checkpoint Inhibitors ◽

Differentially Expressed Gene ◽

Nsclc Cell ◽

Phenotypic Characterization ◽

Nsclc Cell Line

e21015 Background: NSCLC Patients with LKB1 loss respond poorly to immune checkpoint inhibitors (ICI). Determining mechanisms of the underlying immune resistance and strategies to overcome it are urgent and unmet clinical needs. Methods: We re-expressed WT LKB1 into LKB1 mutant NSCLC cell lines and measured effects on immune associated phenotypes in vitro. Specifically, we evaluated response to exogenous interferon gamma (IFNG), as well T-cell mediated cytotoxicity using a T-cell co-culture assay. To identify IFNG signaling changes influenced by LKB1, we evaluated the differential effects of LKB1 on IFNG induced gene expression using RNAseq. Finally, we performed a whole genome loss of function screen using CRISPR-Cas9 library (TKOv3) to identify genes and pathways that modify susceptibility to IFNG in the LKB1 mutant and LKB1 WT state in the A549 cell line. Results: Across multiple LKB1 mutant cell lines, restoration of WT LKB1 enhanced anti-proliferative effects of IFNG in vitro. Furthermore, WT LKB1 enhanced T-cell mediated cytotoxicity in T-cell co-culture assays using anti-NY-ESO TCR and expression of NY-ESO antigen into an HLA matched NSCLC cell line, H2023 (17% vs 40% cell death; P < 0.01). IFNG treatment induced expression of a core set of interferon-driven genes in both the mutant and LKB1 WT state. However, differentially expressed gene classes included downregulation of proliferation-associated genes in the LKB1 WT state, as well as upregulation of ferroptosis genes, and enhanced IFNG-driven expression of immune evasion genes including PDL1, PDL2, TRAF1, and IDO1 in the LKB1 WT state. PDL1 expression was assessed by flow cytometry and found to be upregulated upon expression of LKB1. Our functional genomics screen identified genes whose inhibition enhanced IFNG susceptibility in the A549 mutant state, and genes whose inhibition restored IFNG resistance in the A549 LKB1 WT state. Preliminary analysis identifies candidate signaling and metabolic pathways that appear to confer IFNG sensitivity in the mutant state without significant effect on IFNG sensitivity after LKB1 add-back. These include modifiers of the Hippo/YAP pathway as well as modifiers of ferroptosis, which will form the basis for further mechanistic experiments. Conclusions: ICI resistance caused by LKB1 loss is associated with insensitivity to IFNG, and can be modified in vitro by re-expression of WT LKB1. An integrated approach to evaluate modifiers of IFNG effects identifies resistance mechanisms that may be potential therapeutic targets.

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Ultrastructure of Choriocarcinoma in Vitro

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010006372x ◽

1969 ◽

Vol 27 ◽

pp. 362-363

Author(s):

John C. Garancis ◽

R. A. Pattillo

Keyword(s):

Cell Line ◽

Physiological Function ◽

Cell System ◽

Bewo Cell ◽

Bewo Cells ◽

Gestational Choriocarcinoma ◽

Bewo Cell Line

Growth of cell system (BeWo-cell line) derived from human gestational choriocarcinoma has been established and continuously maintained in-vitro. Furthermore, it is evident from the previous studies that this cell line has retained the physiological function of the placental trophoblasts, namely the synthesis of human chorionic gonadotrophil(HCG).The BeWo cells were relatively small and possessed single nuclei, thus indicating that this cell line consists exclusively of cytotrophoblasts. In some instances cells appeared widely separated and their lateral surfaces were provided with numerous microvilli (Fig.1).

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