Guide to Leukemia-Lymphoma Cell Lines on CD.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4340-4340 ◽  
Author(s):  
Hans G. Drexler ◽  
Roderick A.F. MacLeod ◽  
Stefan Nagel ◽  
Willy G. Dirks ◽  
Cord C. Uphoff ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Molecular Cytogenetics ◽  
Genetic Alterations ◽  
Lymphoma Cell ◽  
Dna Fingerprint ◽  
Human Leukemia ◽  
Continuous Cell Lines ◽  
Functional Features

Abstract The <Guide to Leukemia-Lymphoma Cell Lines> summarizes the salient characteristics of 560 cell lines: precursor B (85), mature B (155), plasma cell leukemia/myeloma (66), immature T (55), mature T (21), natural killer (10), Hodgkin lymphoma (10), anaplastic large cell lymphoma (16), myelocytic (62), monocytic (33) and erythrocytic-megakaryocytic cell lines (45). Along with other improvements, the advent of continuous human leukemia-lymphoma (LL including myeloma) cell lines as a rich resource of abundant, accessible and manipulable living cells has contributed significantly to a better understanding of the pathophysiology of hematopoietic tumors. The first LL cell lines, Burkitt lymphoma-derived lines, were established in 1963. Since then more than 1500 cell lines have been described, some 500 of them in detail. The major advantages of continuous cell lines is the unlimited supply and worldwide availability of identical cell material and the infinite viable storability in liquid nitrogen. LL cell lines are characterized generally by monoclonal origin and differentiation arrest, sustained proliferation in vitro with preservation of most cellular features, and specific genetic alterations characteristic of their tumor of origin. Recent transcriptional profiling studies have confirmed that LL cell lines stably retain the aberrant profiles typical of their tumors of origin. The most practical classification of LL cell lines assigns them to one of the physiologically occurring cell lineages, based on their immunophenotype, genotype and functional features. Truly malignant cell lines should be discerned from Epstein-Barr virus-immortalized normal cells. The efficiency of cell line establishment is rather low and the deliberate establishment of new LL cell lines remains by and large an unpredictable process. Difficulties in establishing continuous cell lines may be caused by the inappropriate selection of nutrients and growth factors. Clearly, a generally suitable microenvironment for hematopoietic cells, either malignant or normal, cannot yet be defined. The characterization and publication of new LL cell lines should provide important and informative core data, attesting to their scientific significance. Large percentages of LL cell lines are contaminated with mycoplasma (about 20%) or are cross-contaminated with other cell lines (14% in our DNA fingerprinting analysis on 622 samples covering 560 LL cell lines). Solutions to these problems are sensitive detection, effective elimination and rigorous prevention of mycoplasma infection and proper, regular authentication of cell lines. The underlying cause appears to be negligent cell culture practice. The willingness of investigators to make their LL cell lines available to others is all too often limited. There is a need in the scientific community for clean and authenticated high quality LL cell lines to which every scientist has access. These are offered by public cell line banks like the DSMZ which holds 169 LL cell lines which all have undergone a strict quality, identity and characterization program (immunoprofile, karyotype, DNA fingerprint, virus/mycoplasma check). An example of the practical utility of LL cell lines are the recent advances in studies of classical and molecular cytogenetics which in large part were made possible by cell lines. We propose a list of the most useful, robust and publicly available reference cell lines which may be used for a variety of experimental purposes. The <Guide to Leukemia-Lymphoma Cell Lines> is available on CD; inquire at <[email protected]>.

scholarly journals CD38 x ICAM1 Bispecific Antibody Is a Novel Approach for Treating Multiple Myeloma and Lymphoma

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 2413-2413
Author(s):  
Xiaocheng Chen ◽  
Oi Kwan Wong ◽  
Leonard Post
Keyword(s):  
Multiple Myeloma ◽  
Cell Line ◽  
Tumor Cells ◽  
Cell Lines ◽  
Adhesion Molecule ◽  
Bispecific Antibody ◽  
Lymphoma Cell ◽  
Cd38 Expression ◽  
Novel Approach

Abstract Targeting cluster of differentiation 38 (CD38) with monoclonal antibodies has resulted in outstanding responses in patients with multiple myeloma (MM). However, a significant portion of patients failed to respond and nearly all patients eventually relapsed. Furthermore, daratumumab, an anti-CD38 antibody, only showed limited monotherapy activity in relapsed/refractory Non-Hodgkin lymphoma (NHL) patients in a phase 2 study. One of the mechanisms of resistance has been partially attributed to lower CD38 expression. Intercellular adhesion molecule 1 (ICAM1), an immunoglobulin (Ig)-like cell adhesion molecule, is highly expressed in multiple myeloma and lymphoma. Antibody against ICAM1, bersanlimab (BI505, BioInvent), was well-tolerated, but only showed limited clinical efficacy in MM patients. Here, we generated bispecific CD38 x ICAM1 antibody to target ICAM1 + tumor types with low to medium CD38 expression. RNA sequencing (RNAseq) results from the Cancer Cell Line Encyclopedia (CCLE) database showed that ICAM1 is highly expressed on myeloma and lymphoma cell lines. ICAM1 expression levels for selected myeloma and lymphoma cell lines were then validated using flow cytometry. The CD38 x ICAM1 bispecific antibody was constructed by paring a novel CD38 antibody and a novel ICAM1 antibody through an asymmetric three chain knob-into-hole format. The bispecific antibody showed potent in vitro antibody-dependent cellular cytotoxicity (ADCC) activities on ICAM1 + tumor cells with medium to low CD38 levels, where daratumumab has low or minimal effect. The bispecific antibody also showed potent in vitro antibody-dependent cellular phagocytosis (ADCP) activities on cell lines with a range of CD38 expression. The CD38 x ICAM1 bispecific antibody further demonstrated potent tumor inhibition activities in in vivo myeloma and lymphoma cell line-derived xenograft (CDX) models, including cell lines with low to medium CD38 expression. We then evaluated CD38 and ICAM1 expressions in lymphoma patient-derived xenograft (PDX) samples by RNAseq. Among the 37 PDX samples, 27 of them showed ICAM1 expression above 2 5 fragments per kilobase of transcript per million map reads (FPKM). On the contrary, there is a wide range of CD38 expression levels with only 6 samples having CD38 expression above 2 5 FPKM. The ICAM1 and CD38 expressions in the selected PDX samples were further validated with IHC staining. Most importantly, the CD38 x ICAM1 bispecific antibody showed complete tumor inhibition in a rituximab-resistant lymphoma PDX model, whereas daratumumab only showed minimal efficacy. In conclusion, the CD38 x ICAM1 bispecific antibody demonstrated improved efficacy and specificity toward CD38 + and ICAM1 + tumor cells and represents a novel approach for treating multiple myeloma and lymphoma. Disclosures No relevant conflicts of interest to declare.

Synthesis and anticancer activity of thiadiazole containing thiourea, benzothiazole and imidazo[2,1-b][1,3,4]thiadiazole scaffolds

2020 ◽  
Vol 16 ◽  
Author(s):  
Stephen Paul Avvaru ◽  
Malleshappa N. Noolvi ◽  
Uttam A More ◽  
Sudipta Chakraborty ◽  
Ashutosh Dash ◽  
...  
Keyword(s):  
Anticancer Activity ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Antitumor Agents ◽  
Data Bank ◽  
Cancer Cell Lines ◽  
Docking Studies ◽  
Human Leukemia

Background: A great array of nitrogen-containing heterocyclic rings were being extensively explored for their functional versatility in the field of medicine especially in anticancer research. 1,3,4-thiadiazole is one of such heterocyclic ring with promising anticancer activity against several cancer cell lines, inhibiting diverse biological targets. Introduction: The 1,3,4-thiadiazole, when equipped with other heterocyclic scaffolds, has displayed enhanced anticancer properties. The thiourea, benzothiazole, imidazo[2,1,b][1,3,4]-thiadiazoles are such potential scaffolds with promising anticancer activity. Method: A new series of 5-substituted-1,3,4-thiadiazoles linked with phenyl thiourea, benzothiazole and 2,6-disubstituted imidazo[2,1- b][1,3,4]thiadiazole derivatives were synthesized and tested for in-vitro anticancer activity on various cancer cell lines. Results: The National Cancer Institute’s preliminary anticancer screening results showed compounds 4b and 5b having potent antileukemic activity. Compound 4b selectively showed 32 percent lethality on Human Leukemia-60 cell line. The docking studies of the derivatives on aromatase enzyme (Protein Data Bank: 3S7S) have shown reversible interactions at the active site with good docking scores comparable to Letrozole and Exemestane. Further, the selected derivatives were tested for anticancer activity on HeLa cell line based on the molecular docking studies. Conclusion: Compound 4b and 5b showed effective inhibition equivalent to Letrozole. These preliminary biological screening studies have given positive anticancer activity for these new classes of derivatives. An additional research study like the mechanism of action of the anticancer activity of this new class of compounds is necessary. These groundwork studies illuminate a future pathway for research of this class of compounds enabling the discovery of potent antitumor agents.

Combination of Enzastaurin, a PKC Inhibitor, and Revlimid ® has Synergistic Activity In Non-Hodgkin Lymphoma

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4905-4905 ◽  
Author(s):  
Stefano Sacchi ◽  
Maria Cosenza ◽  
Monica Civallero ◽  
Giulia Grisendi ◽  
Erika Road ◽  
...  
Keyword(s):  
B Cell ◽  
Cell Line ◽  
Hodgkin Lymphoma ◽  
Cell Lines ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Lymphoma Cell ◽  
Indolent Lymphoma ◽  
Non Hodgkin Lymphoma

Abstract Abstract 4905 A curative treatment does not exist for indolent lymphoma and eventually patients die for progression and complications related to their disease. Thus, there is a need of new less toxic and more active treatment. Enzastaurin, a novel targeted agent, inhibits PKC-β by interacting competitively as its ATP-binding site. Several studies have shown that enzastaurin exhibits growth inhibiting effects on a wide array of cultured human tumour cells. Revlimid ® (lenalidomide), an oral immunomodulatory drug, have shown antineoplastic activity in various tumours, including multiple myeloma (MM), myelodysplastic syndrome (MDS), B-CLL, renal-cell carcinoma and prostate cancer and it is approved for the treatment of patients with MM and MDS bearing a deletion 5q. In the present research, we demonstrate that Revlimid ® alone induce G0/G1 arrest in WSU-NHL cell line, but not apoptosis. This would suggest that, in vitro, Revlimid ® has more a cytostatic than a cytotoxic effect in this cell line. Further, we have demonstrated that the combination of doses as low as 1 mM of Enzastaurin and Revlimid ® exerts, in vitro, a strong synergistic anti lymphoma activity. We also have showed that the combination decreases viability and induce apoptosis in B-cell lymphoma cell lines and peripheral blood mononuclear cells (PBMCs) from follicular lymphoma (FL) patients. The combination has no effect on normal PBMC and suppresses cell proliferation of B-cell lymphoma cell lines when co-cultured with bone marrow stromal cells (BMSCs) in a system that mimics the BM microenvironment. The combination induces a significantly higher rate of apoptosis in comparison with these caused by each agents utilized alone, as showed by flow cytometry. The combination activates both the extrinsic and intrinsic pathways of apoptosis resulting in caspase-8, caspase-9, caspase-3 and PARP cleavage. Furthermore, we have evaluated whether the combination has the ability to trigger apoptosis through BAD and we have showed its ability to activate BAD. Further, we have demonstrated that the combination decreases the expression of phosphorylated AKT and of some AKT downstream targets such as GSK-3β, m-TOR and p70S6. In addition, we have found that the combination reduces the activation of phosphorylated MAPK and of the downstream effector p90RSK. The MAPK signaling pathways have a multiple roles in natural processes such as cell growth, differentiation, and apoptosis. Taken together, these observations suggest that interrupting the PI3K/AKT and MAPK pathways is a promising therapeutic strategy against B-cell lymphoma cell lines. Therefore, these preclinical data, together with promising results obtained with Revlimid ® in the treatment of non-Hodgkin lymphoma, provide the rationale for evaluating the combination of Enzastaurin and Revlimid ® in the treatment of indolent lymphoma. These compounds, with a favourable toxicity profile, are not classic chemotherapeutic agents causing severe side effects and could be considered an example of a new innovative attempt of an anti-cancer “soft treatment”. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Design, Synthesis of Novel Tetrandrine-14-l-Amino Acid and Tetrandrine-14-l-Amino Acid-Urea Derivatives as Potential Anti-Cancer Agents

Molecules ◽  
2020 ◽  
Vol 25 (7) ◽  
pp. 1738
Author(s):  
Sheng-Cao Hu ◽  
Jin Yang ◽  
Chao Chen ◽  
Jun-Rong Song ◽  
Wei-Dong Pan
Keyword(s):  
Amino Acid ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Human Leukemia ◽  
Urea Derivatives ◽  
Anti Cancer

Tetrandrine, a dibenzyltetrahydroisoquinoline alkaloid isolated from the root of the traditional Chinese medicinal plant Stephania tetrandra S. Moore, a member of the Menispermaceae, showed anti-cancer activity by inhibiting cell proliferation, preventing cell cycle progress and induction of cell death and autophagy. In this study, twelve tetrandrine-l-amino acid derivatives and twelve tetrandrine-14-l-amino acid-urea derivatives were designed and synthesized, using C14-aminotetrandrine as raw material. Then the preliminary in vitro anti-cancer activities of these derivatives against human breast cancer cell line MDA-MB-231, human leukemia cell lines HEL and K562 were evaluated. The in vitro cytotoxicity results showed that these derivatives exhibited potent inhibitory effects on cancer cell growth, and the primary structure-activity relationships were evaluated. Notably, compound 3f exhibited satisfactory anticancer activity against all three cancer cell lines, especially the HEL cell line, with the IC50 value of 0.23 µM. Further research showed that 3f could induce G1/S cycle arrest and apoptosis in a dose- and time- dependent manner on the leukemia cell line HEL. The results suggested that 3f may be used as a potential anti-cancer agent for human leukemia.

Treatment with Intermediate Subviral Particles Overcomes Resistance to Reovirus Induced Oncolysis in Mantle Cell Lymphoma Cell Lines.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2417-2417
Author(s):  
Gwyn Bebb ◽  
Huong Muzik ◽  
Tom Alain ◽  
Karen Dong ◽  
Chandini Thirukkamaran ◽  
...  
Keyword(s):  
Mantle Cell Lymphoma ◽  
Cell Line ◽  
Cell Lines ◽  
Cell Lymphoma ◽  
Lymphoma Cell ◽  
Complete Medium ◽  
Mantle Cell ◽  
Time Point ◽  
Subviral Particles

Abstract Introduction: Mantle Cell Lymphoma (MCL), a B-cell, non-Hodgkin lymphoma (NHL) characterized by cyclin D1 and Bcl-2 over-expression, is considered incurable with standard therapies. Reovirus, a double stranded RNA oncolytic virus has shown a consistent anti-neoplastic effect in a variety of cell lines both in vitro, in vivo and ex vivo. We recently demonstrated an anti-lymphoma effect of reovirus in two MCL cell lines and reovirus resistance in three. Here we report the effect of trypsinization of the reovirus envelope to generate intermediate subviral particles (ISVP) on the resistance of these cell lines in vitro. Methods Five MCL cell lines (Z138C, NCEB-1, Hb-12, JVM2 and Granta 519) and one Follicular Lymphoma cell line (DoHH2) were counted and resuspended in serum-free medium. 104 cells in 20ul were dispensed per well in a 96-well plate. 10ul of reovirus at appropriate MOI in serum-medium or 40MOI of ISVP was added to the cells. ISVP was prepared as follows; 1ul of reovirus at 40MOI was treated with 9ul of chymotrypsin at 37°C for 30 minutes before being added to the cells. Cells and viruses were allowed to be in contact at 4°C for 45 min -1 hour. 170 ul of warm complete medium RPMI 1640 containing 10% FCS and L-glutamine was then added and the plate was incubated at 37 °C. Each time point was performed in quadruplicate. WST-1 reagent was used to quantify cell proliferation and cell viability was assessed by absorbance at 450nm using a plate reader. Results MCL cell lines NCEB-1 and JVM-2 showed exquisite sensitivity to reovirus induced oncolysis after 4 days’ incubation at 40 and 80 MOI. Granta exhibited intermediate sensitivity to reovirus while Hb12 and Z138C demonstrated an innate resistance to reovirus induced oncolyis at this time point. Incubation of the resistant cell lines with ISVP overcame this resistance in the Granta and Z138C cell lines (as well as DoHH2). Viability of Granta and Z138C after 4 days’ incubation with ISVP approached that of the sensitive cell lines NCEB-1 and JVM-2 at the same time point. However, the Hb12 cell line retained a significant degree of resistance to reovirus when incubated with ISVP. Percent Viability of 5 MCL Cell Lines at 4 Days’ Incubation with Reovirus or ISVP Treatment NCEB-1 JVM-2 Hb-12 Z138C Granta DoHH2 Control 100 100 100 100 100 100 Reovirus 40 MOI 26 29 113 89 58 54 Reovirus 80 MOI 20 1 114 94 50 45 ISVP 40 MOI 22 5 45 9 12 8 Conclusion Although each of the five cell lines investigated has the karyotypic and molecular hallmarks of MCL, they exhibit heterogeneity of sensitivity to reovirus induced oncolysis. The resistance to reovirus oncolysis exhibited by the two cell lines Z138C and Granta can be overcome by 40 MOI ISVP. In contrast, incubation with the ISVP only partly overcomes reovirus resistance in the HB12 cell line. The molecular basis of this variation and the mechanism underlying reovirus resistance warrants further investigation at the proteome, transcriptome and genome level.

scholarly journals Alternatives for obtaining a continuous cell line from Apis mellifera

2021 ◽  
Vol 51 (12) ◽  
Author(s):  
Matheus Iuri Frühauf ◽  
Lariane da Silva Barcelos ◽  
Nadálin Yandra Botton ◽  
Cristina Mendes Peter ◽  
Silvia de Oliveira Hübner ◽  
...  
Keyword(s):  
Apis Mellifera ◽  
Cell Line ◽  
Cell Lines ◽  
Viral Infections ◽  
Continuous Cell Lines ◽  
Cell Immortalization ◽  
A Cell ◽  
Immortalized Cell ◽  
Choice Of Technique

ABSTRACT: In worldwide there are reports of a significant decrease in colonies of the species Apis mellifera, caused by several factors, including viral infections. In order to study and diagnose illnesses caused by viruses, in vitro cell culture is used as a valuable tool. Yet, there are still no immortalized cell lines of honey bee Apis mellifera. Primary cell cultures are promising for this purpose and can supply the lack of continuous strains, but their establishment is difficult and laborious, which often makes them unfeasible for many research centers. Through the use of cell immortalization techniques, it is possible to develop continuous cell lines and thus benefit, in different ways, research related to different species of bees. The choice of technique is challenging, since in addition to the ability to remain viable for countless passages, cells must keep the genotype and phenotype similar or identical to the original tissue. This review intends to present methodologies that can be used to immortalize Apis mellifera cells, aiming to establish a cell line. The genotypic and phenotypic implications of each technique are evaluated, and the purpose of the cell line to be developed.

ANTITUMOR EFFECT OF SUNITINIB AND BORTEZOMIB ON BREAST CANCER CELL LINE IN VITRO

2017 ◽  
Vol 63 (1) ◽  
pp. 141-145
Author(s):  
Yuliya Khochenkova ◽  
Eliso Solomko ◽  
Oksana Ryabaya ◽  
Yevgeniya Stepanova ◽  
Dmitriy Khochenkov
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Antitumor Effect ◽  
Cancer Cell Lines ◽  
Breast Cancer Cell Lines ◽  
Actual Problem

The discovery for effective combinations of anticancer drugs for treatment for breast cancer is the actual problem in the experimental chemotherapy. In this paper we conducted a study of antitumor effect of the combination of sunitinib and bortezomib against MDA-MB-231 and SKBR-3 breast cancer cell lines in vitro. We found that bortezomib in non-toxic concentrations can potentiate the antitumor activity of sunitinib. MDA-MB-231 cell line has showed great sensitivity to the combination of bortezomib and sunitinib in vitro. Bortezomib and sunitinib caused reduced expression of receptor tyrosine kinases VEGFR1, VEGFR2, PDGFRa, PDGFRß and c-Kit on HER2- and HER2+ breast cancer cell lines

New Mechanistic Insight on the PIM-1 Kinase Inhibitor AZD1208 Using Multidrug Resistant Human Erythroleukemia Cell Lines and Molecular Docking Simulations

2019 ◽  
Vol 19 (11) ◽  
pp. 914-926 ◽  
Author(s):  
Maiara Bernardes Marques ◽  
Michael González-Durruthy ◽  
Bruna Félix da Silva Nornberg ◽  
Bruno Rodrigues Oliveira ◽  
Daniela Volcan Almeida ◽  
...  
Keyword(s):  
Molecular Docking ◽  
Binding Site ◽  
Cell Lines ◽  
Chemotherapeutic Agents ◽  
Atp Binding ◽  
Human Leukemia ◽  
Atp Binding Site ◽  
Mechanistic Insight ◽  
Multiple Drugs

Background:PIM-1 is a kinase which has been related to the oncogenic processes like cell survival, proliferation, and multidrug resistance (MDR). This kinase is known for its ability to phosphorylate the main extrusion pump (ABCB1) related to the MDR phenotype.Objective:In the present work, we tested a new mechanistic insight on the AZD1208 (PIM-1 specific inhibitor) under interaction with chemotherapy agents such as Daunorubicin (DNR) and Vincristine (VCR).Materials and Methods:In order to verify a potential cytotoxic effect based on pharmacological synergism, two MDR cell lines were used: Lucena (resistant to VCR) and FEPS (resistant to DNR), both derived from the K562 non-MDR cell line, by MTT analyses. The activity of Pgp was ascertained by measuring accumulation and the directional flux of Rh123. Furthermore, we performed a molecular docking simulation to delve into the molecular mechanism of PIM-1 alone, and combined with chemotherapeutic agents (VCR and DNR).Results:Our in vitro results have shown that AZD1208 alone decreases cell viability of MDR cells. However, co-exposure of AZD1208 and DNR or VCR reverses this effect. When we analyzed the ABCB1 activity AZD1208 alone was not able to affect the pump extrusion. Differently, co-exposure of AZD1208 and DNR or VCR impaired ABCB1 activity, which could be explained by compensatory expression of abcb1 or other extrusion pumps not analyzed here. Docking analysis showed that AZD1208 is capable of performing hydrophobic interactions with PIM-1 ATP- binding-site residues with stronger interaction-based negative free energy (FEB, kcal/mol) than the ATP itself, mimicking an ATP-competitive inhibitory pattern of interaction. On the same way, VCR and DNR may theoretically interact at the same biophysical environment of AZD1208 and also compete with ATP by the PIM-1 active site. These evidences suggest that AZD1208 may induce pharmacodynamic interaction with VCR and DNR, weakening its cytotoxic potential in the ATP-binding site from PIM-1 observed in the in vitro experiments.Conclusion:Finally, the current results could have a pre-clinical relevance potential in the rational polypharmacology strategies to prevent multiple-drugs resistance in human leukemia cancer therapy.

Design, Synthesis, Molecular Docking, and Anticancer Evaluation of Pyrazole Linked Pyrazoline Derivatives with Carbothioamide Tail as EGFR Kinase Inhibitors

2020 ◽  
Vol 21 (1) ◽  
pp. 42-60
Author(s):  
Farah Nawaz ◽  
Ozair Alam ◽  
Ahmad Perwez ◽  
Moshahid A. Rizvi ◽  
Mohd. Javed Naim ◽  
...  
Keyword(s):  
Molecular Docking ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Cancer Cell Lines ◽  
Kinase Assay ◽  
Cervix Uteri ◽  
Egfr Kinase

Background: The Epidermal Growth Factor Receptor (known as EGFR) induces cell differentiation and proliferation upon activation through the binding of its ligands. Since EGFR is thought to be involved in the development of cancer, the identification of new target inhibitors is the most viable approach, which recently gained momentum as a potential anticancer therapy. Objective: To assess various pyrazole linked pyrazoline derivatives with carbothioamide for EGFR kinase inhibitory as well as anti-proliferative activity against human cancer cell lines viz. A549 (non-small cell lung tumor), MCF-7 (breast cancer cell line), SiHa (cancerous tissues of the cervix uteri), and HCT-116 (colon cancer cell line). Methods: In vitro EGFR kinase assay, in vitro MTT assay, Lactate dehydrogenase release, nuclear staining (DAPI), and flow cytometry cell analysis. Results: Compounds 6h and 6j inhibited EGFR kinase at concentrations of 1.66μM and 1.9μM, respectively. Furthermore, compounds 6h and 6j showed the most potent anti-proliferative results against the A549 KRAS mutation cell line (IC50 = 9.3 & 10.2μM). Through DAPI staining and phase contrast microscopy, it was established that compounds 6h and 6j also induced apoptotic activity in A549 cells. This activity was further confirmed by FACS using Annexin-V-FITC and Propidium Iodide (PI) labeling. Molecular docking studies performed on 6h and 6j suggested that the compounds can bind to the hinge region of ATP binding site of EGFR tyrosine kinase in a similar pose as that of the standard drug gefitinib. Conclusion: The potential anticancer activity of compounds 6h and 6j was confirmed and need further exploration in cancer cell lines of different tissue origin and signaling pathways, as well as in animal models of cancer development.

Survival and Function of Fibroblasts Stored as Stock Cultures at a Non-freezing Temperature

1993 ◽  
Vol 21 (2) ◽  
pp. 206-209
Author(s):  
Anders H. G. Andrén ◽  
Anders P. Wieslander
Keyword(s):  
Cell Growth ◽  
Cell Line ◽  
Cell Lines ◽  
Fibroblast Cell ◽  
Freezing Temperature ◽  
Mouse Fibroblast ◽  
Toxic Response ◽  
Normal Manner ◽  
And Function

Cytotoxicity, measured as inhibition of cell growth of cultured cell lines, is a widely used method for testing the safety of biomaterials and chemicals. One major technical disadvantage with this method is the continuous routine maintenance of the cell lines. We decided to investigate the possibility of storing stock cultures of fibroblasts (L-929) in an ordinary refrigerator as a means of reducing the routine workload. Stock cultures of the mouse fibroblast cell line L-929 were prepared in plastic vials with Eagle's minimum essential medium. The vials were stored in a refrigerator at 4–10°C for periods of 7–31 days. The condition of the cells after storage was determined as cell viability, cell growth and the toxic response to acrylamide, measured as cell growth inhibition. We found that the L-929 cell line can be stored for 2–3, weeks with a viabilty > 90% and a cell growth of about 95%, compared to L-929 cells grown and subcultured in the normal manner. The results also show that the toxic response to acrylamide, using refrigerator stored L-929 cells, corresponds to that of control L-929 cells. We concluded that it is possible to store L-929 cells in a refrigerator for periods of up to 3 weeks and still use the cells for in vitro cytotoxic assays.

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