scholarly journals Identification of the paired basic convertases implicated in HIV gp160 processing based on in vitro assays and expression in CD4+ cell lines.

1997 ◽  
Vol 272 (13) ◽  
pp. 8836
Author(s):  
Etienne Decroly ◽  
Sandrine Wouters ◽  
Carlo Di Bello ◽  
Claude Lazure ◽  
Jean-Marie Ruysschaert ◽  
...  
Keyword(s):  
Cell Lines ◽  
In Vitro Assays ◽  
Cd4 Cell

TMOD-31. AN ORGANOTYPIC TISSUE PLATFORM TO BRIDGE IN VITRO AND IN VIVO ASSAYS FOR BRAIN CANCER TREATMENT

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi222-vi222
Author(s):  
Breanna Mann ◽  
Noah Bell ◽  
Denise Dunn ◽  
Scott Floyd ◽  
Shawn Hingtgen ◽  
...  
Keyword(s):  
Cell Lines ◽  
Brain Cancer ◽  
Brain Slice ◽  
Brain Slices ◽  
In Vitro Assays ◽  
Preclinical Model ◽  
Short Period ◽  
The Brain

Abstract Brain cancers remain one of the greatest medical challenges. The lack of experimentally tractable models that recapitulate brain structure/function represents a major impediment. Platforms that enable functional testing in high-fidelity models are urgently needed to accelerate the identification and translation of therapies to improve outcomes for patients suffering from brain cancer. In vitro assays are often too simple and artificial while in vivo studies can be time-intensive and complicated. Our live, organotypic brain slice platform can be used to seed and grow brain cancer cell lines, allowing us to bridge the existing gap in models. These tumors can rapidly establish within the brain slice microenvironment, and morphologic features of the tumor can be seen within a short period of time. The growth, migration, and treatment dynamics of tumors seen on the slices recapitulate what is observed in vivo yet is missed by in vitro models. Additionally, the brain slice platform allows for the dual seeding of different cell lines to simulate characteristics of heterogeneous tumors. Furthermore, live brain slices with embedded tumor can be generated from tumor-bearing mice. This method allows us to quantify tumor burden more effectively and allows for treatment and retreatment of the slices to understand treatment response and resistance that may occur in vivo. This brain slice platform lays the groundwork for a new clinically relevant preclinical model which provides physiologically relevant answers in a short amount of time leading to an acceleration of therapeutic translation.

scholarly journals Evaluation of the Cytopathicity (Fusion/Hemifusion) of Patient-Derived HIV-1 Envelope Glycoproteins Comparing Two Effector Cell Lines

2012 ◽  
Vol 17 (6) ◽  
pp. 727-737 ◽  
Author(s):  
Francesc Cunyat ◽  
Marta Curriu ◽  
Silvia Marfil ◽  
Elisabet García ◽  
Bonaventura Clotet ◽  
...  
Keyword(s):  
Cell Death ◽  
Single Cell ◽  
Cell Lines ◽  
Effector Cell ◽  
Cell Loss ◽  
Biological Properties ◽  
In Vitro Assays ◽  
Envelope Glycoproteins ◽  
Hiv 1

HIV-1 envelope glycoprotein (Env) is a major determinant of viral pathogenicity. The evaluation of the biological properties of patient-derived envelopes by comparing two effector cell lines (293T and HeLa) is reported. A standard cell-to-cell fusion assay was used to evaluate fusogenicity, whereas a coculture with CD4+ cells was used to evaluate absolute cell loss, single cell death, and hemifusion events. Fusion and absolute cell loss assays showed that Env-expressing 293T and HeLa cells had different fusion efficiencies; fusion was magnified in 293T cells despite a significantly lower cell-surface Env expression. Conversely, gp41-mediated single cell death and hemifusion induced in CD4+ cells by 293T-Env-positive cells were significantly lower than that induced by HeLa-Env-positive cells. These data showed that the effector cell line used in the in vitro assays is crucial, and a combination of assays is recommended to evaluate the biological properties of patient-derived envelope glycoproteins: preferentially, 293T-Env-positive cells for the evaluation of fusogenicity and HeLa-Env-positive cells for the evaluation of cell death parameters. The combination of assays described in our work could be a valuable tool for dual screenings of large collections of primary Envs or Env mutants and drugs acting on these Envs.

scholarly journals Identification of a Potent Cytotoxic Pyrazole with Anti-Breast Cancer Activity That Alters Multiple Pathways

Cells ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 254
Author(s):  
Denisse A. Gutierrez ◽  
Lisett Contreras ◽  
Paulina J. Villanueva ◽  
Edgar A. Borrego ◽  
Karla Morán-Santibañez ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Lines ◽  
Human Cancer ◽  
Stress Protein ◽  
In Vitro Assays ◽  
Parp Cleavage ◽  
Human Cancer Cell Lines ◽  
Microtubule Disruption ◽  
Tnbc Cell

In this study, we identified a novel pyrazole-based derivative (P3C) that displayed potent cytotoxicity against 27 human cancer cell lines derived from different tissue origins with 50% cytotoxic concentrations (CC50) in the low micromolar and nanomolar range, particularly in two triple-negative breast cancer (TNBC) cell lines (from 0.25 to 0.49 µM). In vitro assays revealed that P3C induces reactive oxygen species (ROS) accumulation leading to mitochondrial depolarization and caspase-3/7 and -8 activation, suggesting the participation of both the intrinsic and extrinsic apoptotic pathways. P3C caused microtubule disruption, phosphatidylserine externalization, PARP cleavage, DNA fragmentation, and cell cycle arrest on TNBC cells. In addition, P3C triggered dephosphorylation of CREB, p38, ERK, STAT3, and Fyn, and hyperphosphorylation of JNK and NF-kB in TNBC cells, indicating the inactivation of both p38MAPK/STAT3 and ERK1/2/CREB signaling pathways. In support of our in vitro assays, transcriptome analyses of two distinct TNBC cell lines (MDA-MB-231 and MDA-MB-468 cells) treated with P3C revealed 28 genes similarly affected by the treatment implicated in apoptosis, oxidative stress, protein kinase modulation, and microtubule stability.

Canonical NF-kB Pathway Is Not Constitutively Activated and May Not Be the Major Bortezomib Target in Mantle Cell Lymphoma (MCL).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2591-2591
Author(s):  
Valerie Camara-Clayette ◽  
Yves Lepelletier ◽  
Ivan Moura ◽  
William Vainchenker ◽  
Olivier Hermine ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Line ◽  
Cell Lines ◽  
Myeloma Cell ◽  
Dominant Negative ◽  
In Vitro Assays ◽  
Lymphoid Malignancies ◽  
Empty Vector ◽  
Gene Assay

Abstract Bortezomib, a proteasome inhibitor, has promising activity in MCL. Bortezomib mechanism of action is complex since multiple proteins and cellular pathways are regulated by proteasome degradation. By blocking the degradation of IKBa, the canonical NF-kB pathway is one of the major Bortezomib target in lymphoid malignancies such as multiple myeloma. NF-kB activation has not been extensively assessed in other lymphoid malignancies such as MCL. NF-kB complexes (heterodimers P50/P65) were assessed by EMSA assays in REC GRANTA 519, NCEB, JEKO, JUN and UPN1 MCL cell-lines. EBV was present in 3/6 cell lines. P50/P65 complexes were detected in all EBV positive cell lines and only in 1/3 EBV negative cell-lines (JEKO). No nuclear P65 protein was detected by immunofluorescence or Western-blot analysis in the 2 other negative EBV cell lines (REC and UPN1). NF-kB transcriptional activity was measured by a luciferase-based reporter gene assay. Spontaneous NF-kB activity was low in UPN1, REC and JEKO compared to GRANTA 519 EBV positive cell-line (3 to 4 time lower) but can be strongly activated (up to 10 fold) using MEKK cotransfection assay. This suggest that NF-kB is functional, but not constitutively activated. Stable infections with Migr1-IRES-GFP IkBM (dominant negative IKB) or empty vector, were performed in GRANTA 519 EBV positive as well as in UPN1 cell line. No differences in proliferation or apoptosis were observed in UPN1 stably infected with IkBM or with the empty vector but GRANTA 519 EBV positive showed increased apoptosis and proliferation inhibition when infected with IkBM. Likewise, no P50/P65 heterodimers complexes were detected by EMSA in 4 patients with MCL. In vitro assays showed that UPN1 and JEKO cell lines had comparable sensitivities to Bortezomib than Multiple myeloma cell lines reported in the literature (IC 50: 6 nM and 12 nM respectively). This was also true for the 3 patients lymphoma cells assessed in vitro (12 nM). Taken together these results suggest that canonical NF-kB activation pathway is not constitutively active in EBV negative MCL cell lines and patients samples and could be associated with EBV infection in some MCL cell-lines. Therefore, this strongly suggests that Bortezomib target other molecules in MCL.

Pharmaco-Toxicological Study of Kageneckia oblonga, Rosaceae

2002 ◽  
Vol 57 (1-2) ◽  
pp. 100-108 ◽  
Author(s):  
Carla Delporte ◽  
Orlando Muñozb ◽  
Javier Rojas ◽  
Marisa Ferrándiz ◽  
Miguel Payá ◽  
...  
Keyword(s):  
Cell Lines ◽  
Lung Carcinoma ◽  
Methanol Extract ◽  
Human Lung ◽  
Antioxidant Properties ◽  
In Vitro Assays ◽  
Human Neutrophils ◽  
Human Lung Carcinoma ◽  
Ht 29

The probable antipyretic, antiinflammatory, analgesic and antioxidant properties of Kageneckia oblonga, Rosaceae, were investigated and the major compounds of its active extracts were isolated. The study comprised the acute toxicity of the extracts of global methanol, hexane, dichloromethane and methanol. The cytotoxicity of global methanol extract was studied in three tumoral cell lines. All the extracts exhibited the pharmacological activities under study. Methanol and dichloromethane were the most toxic extracts. From the global methanol extract, isolations were performed of prunasin, 23,24- dihydro-cucurbitacin F, and a new cucurbitacin, 3β-(β-d-glucosyloxy)-16α,23α-epoxycucurbita-5,24-diene-11-one. The cytotoxicity of both cucurbitacins on human neutrophils at the assayed concentrations was not statistically significant. In-vitro assays showed that both cucurbitacins can be partly responsible for the analgesic, antipyretic, and anti-inflammatory activities. Evaluation was done of the cytotoxicity of global methanol extract, 23, 24-dihydrocucurbitacin F, aqueous extracts and prunasin against P-388 murine leukaemia, A-549 human lung carcinoma and HT-29 colon carcinoma. Since global methanol extract presented a strong cytotoxicity against P-388 murine leukaemia, A-549 human lung carcinoma, and HT-29 cell lines, it is highly probable that this extract contain one or more cytotoxic compounds that could be investigated for their potential use as an agent against cancer.

scholarly journals Synthesis and Biological Screening of New Cyano-Substituted Pyrrole Fused (Iso)Quinoline Derivatives

Molecules ◽  
2021 ◽  
Vol 26 (7) ◽  
pp. 2066
Author(s):  
Maria Cristina Al-Matarneh ◽  
Roxana-Maria Amărandi ◽  
Ionel I. Mangalagiu ◽  
Ramona Danac
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Human Cancer ◽  
Cancer Cell Lines ◽  
In Vitro Assays ◽  
Biological Screening ◽  
Human Cancer Cell Lines ◽  
Breast And Prostate Cancer

Several new cyano-substituted derivatives with pyrrolo[1,2-a]quinoline and pyrrolo[2,1-a]isoquinoline scaffolds were synthesized by the [3 + 2] cycloaddition of (iso)quinolinium ylides to fumaronitrile. The cycloimmonium ylides reacted in situ as 1,3-dipoles with fumaronitrile to selectively form distinct final compounds, depending on the structure of the (iso)quinolinium salt. Eleven compounds were evaluated for their anticancer activity against a panel of 60 human cancer cell lines. The most potent compound 9a showed a broad spectrum of antiproliferative activity against cancer cell lines representing leukemia, melanoma and cancer of lung, colon, central nervous system, ovary, kidney, breast and prostate cancer. In vitro assays and molecular docking revealed tubulin interaction properties of compound 9a.

Comparative Sensitivity of Two Mouse Lymphoid Tumour Cell Lines (P388 and P388D1) to Five Anticancer Drugs In Vitro

1990 ◽  
Vol 18 (1_part_1) ◽  
pp. 225-230
Author(s):  
Roland Arnould ◽  
Jacque Dubois ◽  
Fokri Abikhalil ◽  
Anita Libert ◽  
Ghanem Ghanem ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Tritiated Thymidine ◽  
In Vitro Assays ◽  
Lower Sensitivity ◽  
Thymidine Uptake ◽  
Tritiated Thymidine Uptake ◽  
P388 Cell ◽  
Repeated Passage

The sensitivity and the selectivity of two murine lymphoid cell lines (P388 and P388D1) to five chemotherapeutic drugs were investigated in vitro. The cytotoxicities of melphalan, daunorubicin, hexamethylmelamine (HMM), hydroxymethylpentamethylmelamine and dihydroxymethyltetramethylmelamine, two HMM derivatives, were measured in the two cell lines using two different techniques: reduction of a tetrazolium derivative (MTT) and tritiated thymidine uptake into DNA. Cytotoxicity was expressed by the 50% inhibitory concentration (IC50) after one hour and after exposure of cells to each drug for two days. The IC50 results indicate that the P388 cells were generally more sensitive to melphalan, daunorubicin, hydroxymethylpentamethylmelamine and dihydroxymethyltetramethylmelamine than the P388D1 cell line. HMM was found to be inactive in both cell lines. Despite the lower sensitivity of the P388D1 cell line compared with the P388 cell line, because of its greater homogeneity, it could replace the P388 line for the in vitro assays, bearing in mind that the P388D1 cell line sensitivity is 1.21 - 24.63 times lower than that of the P388 cell line, at least as far as the drugs tested are concerned. Moreover, our results emphasise that variations of sensitivity could occur with repeated passage in vitro.

scholarly journals Establishment of a Novel In Vitro Model of Endometriosis with Oncogenic KRAS and PIK3CA Mutations for Understanding the Underlying Biology and Molecular Pathogenesis

Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3174
Author(s):  
Mohammad Mahmud Hossain ◽  
Kentaro Nakayama ◽  
Kamrunnahar Shanta ◽  
Sultana Razia ◽  
Masako Ishikawa ◽  
...  
Keyword(s):  
Cell Lines ◽  
Lysyl Oxidase ◽  
Mutant Cell ◽  
Molecular Pathogenesis ◽  
In Vitro Assays ◽  
Epithelial Cell Line ◽  
Pik3ca Mutations ◽  
Invasion And Migration ◽  
And Migration

Endometriosis-harboring cancer-associated somatic mutations of PIK3CA and KRAS provides new opportunities for studying the multistep processes responsible for the functional and molecular changes in this disease. We aimed to establish a novel in vitro endometriosis model to clarify the functional behavior and molecular pathogenesis of this disorder. Immortalized HMOsisEC10 human ovarian endometriotic epithelial cell line was used in which KRAS and PIK3CA mutations were introduced. Migration, invasion, proliferation, and microarray analyses were performed using KRAS and PIK3CA mutant cell lines. In vitro assays showed that migration, invasion, and proliferation were significantly increased in KRAS and PIK3CA mutant cell lines, indicating that these mutations played causative roles in the aggressive behavior of endometriosis. Microarray analysis identified a cluster of gene signatures; among them, two significantly upregulated cancer-related genes, lysyl oxidase (LOX) and pentraxin3 (PTX3), were associated with cell proliferation, invasion, and migration capabilities. Furthermore, siRNA knockdown of the two genes markedly reduced the metastatic ability of the cells. These results suggest that endometriosis with KRAS or PIK3CA mutations can significantly enhance cell migration, invasion, and proliferation by upregulating LOX and PTX3. We propose that LOX and PTX3 silencing using small molecules could be an alternative therapeutic regimen for severe endometriosis.

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