Difference of Drug-Resistance between Single-Factor Resistance Cell Line K562/MDR1 Transfected with mdr1 Gene and Multi-Factor Resistance Cell Line K562/A02 Induced by Doxorubicin.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4179-4179
Author(s):  
Yuping Gong ◽  
Xueshi Ye ◽  
Ting Liu
Keyword(s):  
Drug Resistance ◽  
Cell Line ◽  
Cell Lines ◽  
Anticancer Drug ◽  
Leukemia Cell Line ◽  
Mdr1 Gene ◽  
Sensitive Cell ◽  
Single Factor ◽  
Efflux Rate ◽  
Sensitive Cell Line

Abstract Objective To study difference of drug-resistance between single-factor resistance cell line K562/MDR1 transfected with mdr1 gene and multi-factor resistance cell line K562/A02 induced by doxorubicin. Methods Retroviral virions carrying the complete sequence of mdr1 gene cDNA were produced and infected drug-sensitive leukemia cell line K562 and mdr1 single-factor resistance cell line K562/MDR1 was established. The difference of drug-resistance between K562/MDR1 and K562/A02, a kind of multi-factor resistance cell line induced by doxorubicin, was studied by checking the expression of mdr1 gene and Pgp, daunorubicin efflux rate, MTT drug sensitivity to chemotherapeutic drug. Lentiviral vector encoding shRNA which targeted MDR1 gene was transfected into two kinds of cell lines and effect of RNAi on reversing drug resistance was detected. Results The results of Q-PCR and flow cytometry demonstrated that there were high expression of mdr1 mRNA and Pgp in both kinds of drug-resistance cell lines and no difference between them. The function of Pgp detected by daunorubicin efflux rate is higher in K562/MDR1 (90.93%) than K562/A02 (78.67%). The results of MTT test showed that IC50 of K562/MDR1 and K562/A02 is 0.55 and 1.22μmol/L respectively and this confirmed that drug-resistance in K562/A02 is higher than that in K562/MDR1. After RNA interference, the expression of the mdr1 gene and Pgp in K562/MDR1 markedly was down-regulated and the drug resistance was restored and IC50 is 0.16μmol/L, similar to K562 sensitive cell line. The expression of the mdr1 gene and Pgp in K562/A02 markedly was downregulated too, and drug resistance to anticancer drug is reduced to some extent but IC50 is 0.56μmol/L, it is still higher than that in sensitive cell line. Conclusion Drug-resistance in K562/A02 induced by anticancer-drug was made of many factors and it is more resistance to anticancer-drug than that in K562/MDR1 caused by mdr1 gene. Due to only mdr1 resistance, K562/MDR1 is better cell model to make mdr1/Pgp research.

MDR1 gene overexpression confers resistance to imatinib mesylate in leukemia cell line models

Blood ◽  
2003 ◽  
Vol 101 (6) ◽  
pp. 2368-2373 ◽  
Author(s):  
François-Xavier Mahon ◽  
Francis Belloc ◽  
Valérie Lagarde ◽  
Claudine Chollet ◽  
François Moreau-Gaudry ◽  
...  
Keyword(s):  
Cell Line ◽  
Imatinib Mesylate ◽  
Cell Lines ◽  
Flow Cytometric Analysis ◽  
Leukemia Cell ◽  
Differential Sensitivity ◽  
Leukemia Cell Line ◽  
Mdr1 Gene ◽  
Gene Overexpression ◽  
Clinical Resistance

Inappropriate expression of the multidrug resistance(MDR1) gene encoding the P-glycoprotein (Pgp) has been frequently implicated in resistance to different chemotherapeutic drugs. We have previously generated chronic myeloid leukemia (CML) cell lines resistant to the tyrosine kinase inhibitor imatinib mesylate (STI571), and one line (LAMA84-r) showed overexpression not only of the Bcr-Abl protein but also of Pgp. In the present study, we investigated this phenomenon in other cell lines overexpressing exclusively Pgp. Thus, cells from the K562/DOX line, described as resistant to doxorubicin due to MDR1 gene overexpression, grew continuously in the presence of 1 μM imatinib, but died in 4 to 5 days if the Pgp pump modulators verapamil or PSC833 were added to the imatinib-treated culture. Analysis of cell proliferation by the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay confirmed the differential sensitivity of K562/DOX to imatinib, which was also reversed by verapamil or PSC833. Flow cytometric analysis of the total phosphotyrosine content by intracytoplasmic staining after a 2-hour incubation with escalating doses of imatinib showed that the inhibitory concentrations of 50% (IC50) for inhibition of cellular protein tyrosine phosphorylation were 15, 10, and 5 μM for K562/DOX, K562/DOX plus verapamil, and K562, respectively. Retroviral-mediated transfection of theBCR-ABL+ AR230 cell line with theMDR1 gene decreased its sensitivity to imatinib, an effect that was also reversed by verapamil. The possible role of MDR overexpression in clinical resistance to imatinib remains to be defined. We therefore confirm that imatinib should be added to the extensive list of drugs that can be affected by the MDR phenomenon.

Changes in gene expression profile in two multidrug resistant cell lines derived from a same drug sensitive cell line

2014 ◽  
Vol 38 (8) ◽  
pp. 983-987 ◽  
Author(s):  
Miguel Angelo Martins Moreira ◽  
Carolina Bagni ◽  
Marcos Barcelos de Pinho ◽  
Thaís Messias Mac-Cormick ◽  
Mateus dos Santos Mota ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Gene Expression Profile ◽  
Cell Lines ◽  
Expression Profile ◽  
Multidrug Resistant ◽  
Resistant Cell ◽  
Sensitive Cell ◽  
Sensitive Cell Line

Downregulation of Survivin by Arsenic Trioxide in Acute Myeloid Leukemia Cell Line.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4384-4384
Author(s):  
Karina Lani Silva ◽  
Martina de Freitas Prazeres ◽  
Raquel Ciuvalschi Maia
Keyword(s):  
Drug Resistance ◽  
Cell Line ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Leukemia Cell ◽  
Survivin Expression ◽  
Annexin V ◽  
Leukemia Cell Line ◽  
Acute Myeloid ◽  
Myeloid Leukemia Cell

Abstract Caspases are proteins that play a central role in apoptosis. Therefore, triggering apoptosis through chemotherapeutical caspases inductor drugs is the major path in cancer treatment. However, hindering apoptosis by inhibitor of apoptosis proteins (IAPs) overexpression, have been described in many cancer types including leukemia and, is frequently related to drug resistance. Survivin, a member of IAPs family, is expressed in most human cancers but undetectable in the majority of normal adult tissues. In acute myeloid leukemia (AML), Survivin expression has been correlated with poor prognosis and chemotherapy resistance. These characteristics make Survivin eligible for a promising target for AML treatment. To explore the relationship between Survivin and drug resistance we investigated the alteration of Survivin expression in two AML cell lines HL60 (AML-M2) and U937 (AML-M5) and one chronic myeloid leukemia cell line in blast crisis for M6 (K562) treated with two chemotherapeutic drugs used in leukemia treatment: arsenic trioxide (As2O3) and doxorubicin (Dox). MTT assay was performed to determine the dose of drugs capable to induce cell death in 50% of treated cells (DL50). To verify the percentage of apoptosis induced by As2O3 and Dox at DL50 concentrations determined by MTT, the annexin V/propidium iodide-staining assay was performed and analyzed by flow cytometer. Western blot was used to analyze Survivin expression before and after drugs treatment at DL50 concentrations. Among the cell lines studied, HL60 was the most sensitive for both drugs tested. The DL50 concentrations obtained for As2O3 were 2, 4 and 5 μM at 24 hours for HL60, U937 and K562, respectively. Dox DL50 concentrations were 10 μM at 24 hours for HL60, 5 μM at 48 hours for K562 and 1 μM at 72 hours for U937. The annexin-V/PI staining showed that these drugs were capable to induce apoptosis in all cell lines tested. The percentages of apoptosis induction for As2O3 were 50% for HL60, 21,84% for U937 and 32,7% for K562 in comparison with control cells, while for Dox, the index of apoptosis were 86,8%, 35,7% and 2,2% for HL60, U937 and K562, respectively. Interestingly, at DL50 concentrations As2O3 and Dox inhibited Survivin expression in 72,7% and 69,2%, respectively. No significant alteration in Survivin expression was observed in U937 and K562 cell lines. Thus, HL60 cell line was the most sensible cell line studied and it was correlated with downregulation of Survivin expression. It suggests that Survivin could be considered a therapeutic target for AML-M2 and that As2O3 and Dox are capable to decrease drug resistance. However, the mechanism of action of As2O3 and Dox in Survivin expression seems to be cell type specific.

Simultaneous Inhibition of cIAP1, cIAP2 and XIAP Is Required for Inducing Apoptosis in Multiple Myeloma Cells.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2943-2943
Author(s):  
Vijay G. Ramakrishnan ◽  
Teresa K. Kimlinger ◽  
Utkarsh Painuly ◽  
Jessica Haug ◽  
S. Vincent Rajkumar ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Signaling Pathways ◽  
Cell Line ◽  
Cell Lines ◽  
Research Funding ◽  
Sensitive Cell ◽  
Sensitive Cell Line ◽  
Nfkb Pathway ◽  
Stat Pathway

Abstract Abstract 2943 Background: Inhibitor of apoptosis (IAP) proteins represents a conserved group of proteins that are important regulators of cell survival and apoptosis. X-linked IAP (XIAP) is the best studied IAP that inhibits pro-apoptotic caspases 3, 7 and 9. Multiple myeloma (MM) cell lines express high levels of XIAP. The levels of XIAP are further increased when stimulated by cytokines IL6 and IGF-1, both secreted in copious amounts in myeloma microenvironment. The other two main IAP proteins, namely cIAP1 and cIAP2 are not direct inhibitors of caspases. Instead, they modulate the levels of various signaling pathways by ubiquitinating proteins within the pathways. The NFKB pathway could be activated or inhibited by cIAP1 and 2. In MM, deletions of cIAP1 and cIAP2 have been shown to activate non-canonical NFKB pathway, which indicates a possible tumor suppressor role of these proteins. We wanted to investigate the role of the three IAPs by using a small molecule inhibitor. Our studies clearly indicate the importance of inhibiting all the three IAPs for the induction of apoptosis in MM cells. Methods: LCL161 was synthesized by Novartis Inc. (Basel, Switzerland). Stock solutions were made in DMSO, and subsequently diluted in RPMI-1640 medium for use. MM cell lines were cultured in RPMI 1640 containing 10% fetal bovine serum (20% serum for primary patient cells) supplemented with L-Glutamine, penicillin, and streptomycin. Cytotoxicity was measured using the MTT viability assay and proliferation using thymidine uptake. Apoptosis was measured using flow cytometry with Annexin V-FITC and propidium iodide (PI) for cell lines and patient cells. Immunoblotting was done on cell extracts at various time points following incubation with the drug in order to study the cell signaling pathways. siRNA to cIAP2 was purchased from Invitrogen and was electroporated into MM1S cells. Results: We first examined baseline levels of cIAP1, cIAP2 and XIAP in several MM cell lines and a few patient cells. We observed that the IAPs were constitutively expressed in MM cells. We then wanted to examine the functional significance of these IAP proteins in MM cells. For this, we used an IAP inhibitor LCL161. We observed that LCL161 was able to induce cytotoxicity and inhibit proliferation of MM cells, albeit with differences observed between cell lines. We then examined the factors contributing to resistance in the less sensitive cell lines. For this we chose H929, a sensitive cell line and MM1S, a less sensitive cell line to LCL161. Upon treatment with LCL161, cIAP1 and XIAP were down regulated accompanied by increase in levels of activated caspases 9, 8 and 3 in both H929 and MM1S cells. Using LCL161 in combination with a caspase 9 or a caspase 8 or a pan caspase inhibitor showed clearly that the extrinsic pathway is more involved in the LCL161 induced cell death process. LCL161, however, was unable to inhibit cIAP2 in the less sensitive cell line MM1S whereas cIAP2 was not found to be expressed in H929 cells. It has been shown that cIAP1 is required for ubiquitination and degradation of cIAP2. Therefore, cIAP1 down regulation by LCL161 could actually be contributing to the lack of down regulation of cIAP2 and the observed resistance to LCL161. In order to test this, we used a siRNA to cIAP2 and transfected it into MM1S cells by electroporation. We observed that the siRNA reduced cIAP2 levels and in combination with LCL161 led to marked increase in cells undergoing apoptosis. We also examined signaling pathways after treatment with LCL161 and observed upregulation of both canonical and non-canonical NFKB pathways and Jak/Stat pathway in MM1S cells and not in H929 cells. Combining LCL161 with a Jak2 specific inhibitor SD-1029 synergized in inducing cell death in MM1S and other cell lines less sensitive to LCL161. We are currently testing this combination in MM patient cells. Conclusion: These studies demonstrate the importance of inhibiting cIAP1, cIAP2 and XIAP together in MM cells. Furthermore, by this study we were able to identify resistance mechanisms that are upregulated due to inhibiting the IAP proteins and the importance of using agents that inhibit the IAPs along with inhibitors of these pathways in inducing apoptosis in MM cells. The findings from these studies form the basis of evaluation of IAP inhibitors in combination with a Jak/Stat pathway inhibitor in patients with MM. Disclosures: Kumar: Celgene: Consultancy, Research Funding; Merck: Consultancy, Honoraria; Millennium Pharmaceuticals, Inc.: Research Funding; Novartis: Research Funding; Genzyme: Research Funding; Cephalon: Research Funding.

scholarly journals Biological response and cytotoxicity induced by lipid nanocapsules

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Marzena Szwed ◽  
Maria Lyngaas Torgersen ◽  
Remya Valsala Kumari ◽  
Sunil Kumar Yadava ◽  
Sascha Pust ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Fate ◽  
Cell Lines ◽  
Breast Cancer Cell Lines ◽  
Minor Effect ◽  
Sensitive Cell ◽  
Sensitive Cell Line ◽  
Lipid Nanocapsules ◽  
Knock Down ◽  
Mcf 7

Abstract Background Lipid nanocapsules (LNCs) are promising vehicles for drug delivery. However, since not much was known about cellular toxicity of these nanoparticles in themselves, we have here investigated the mechanisms involved in LNC-induced intoxication of the three breast cancer cell lines MCF-7, MDA-MD-231 and MDA-MB-468. The LNCs used were made of Labrafac™ Lipophile WL1349, Lipoid® S75 and Solutol® HS15. Results High resolution SIM microscopy showed that the DiD-labeled LNCs ended up in lysosomes close to the membrane. Empty LNCs, i.e. without encapsulated drug, induced not only increased lysosomal pH, but also acidification of the cytosol and a rapid inhibition of protein synthesis. The cytotoxicity of the LNCs were measured for up to 72 h of incubation using the MTT assay and ATP measurements in all three cell lines, and revealed that MDA-MB-468 was the most sensitive cell line and MCF-7 the least sensitive cell line to these LNCs. The LNCs induced generation of reactive free oxygen species and lipid peroxidation. Experiments with knock-down of kinases in the near-haploid cell line HAP1 indicated that the kinase HRI is essential for the observed phosphorylation of eIF2α. Nrf2 and ATF4 seem to play a protective role against the LNCs in MDA-MB-231 cells, as knock-down of these factors sensitizes the cells to the LNCs. This is in contrast to MCF-7 cells where the knock-down of these factors had a minor effect on the toxicity of the LNCs. Inhibitors of ferroptosis provided a large protection against LNC toxicity in MDA-MB-231 cells, but not in MCF-7 cells. Conclusions High doses of LNCs showed a different degree of toxicity on the three cell lines studied, i.e. MCF-7, MDA-MD-231 and MDA-MB-468 and affected signaling factors and the cell fate differently in these cell lines.

Enhanced DNA Repair Via Fanconi Anemia/BRCA Pathway Is Involved in Melphalan-Resistant Myeloma Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 284-284
Author(s):  
Qing Chen ◽  
Pieter C. Van der Sluis ◽  
Lori Hazlehurst ◽  
William S. Dalton
Keyword(s):  
Drug Resistance ◽  
Dna Repair ◽  
Cell Line ◽  
Fanconi Anemia ◽  
Resistant Cell ◽  
Sensitive Cell ◽  
Cross Link ◽  
Sensitive Cell Line ◽  
Repair Capacity ◽  
Myeloma Cells

Abstract Melphalan, a DNA crosslinker, is one of the most widely used and effective drugs in the treatment of multiple myeloma (MM). Interstrand cross-links (ICL) are amongst the most toxic types of DNA damage; therefore, DNA cross-linking agents are important drugs in cancer treatment. Unfortunately, although most patients respond to standard and high dose melphalan therapy, eventually patients acquire drug resistance. Acquired melphalan resistance has been associated with reduced DNA crosslinks, elevated levels of glutathione and increased radiation survival. However, mechanisms associated with resistance are not well understood. Evidence has accumulated to suggest that ICL repair contributes to the melphalan resistance. In this study, we compared the gene expression profile (GEP) of the melphalan-resistant myeloma cell line, 8226/LR5 to the 8226/S drug sensitive cell line, and found genes involved in FANC/BRCA DNA cross-link repair pathway had increased expression in drug resistant cells. The aim of our study was to determine whether FANC-BRCA pathway affects the DNA cross link repair capacity and accounts for acquired melphalan-resistance. Using real time RT-PCR and Western Blotting, we examined the expression levels of FANC/BRCA pathway genes in two different drug sensitive and resistant cell lines: 8226/S 8226/LR5, and U266/S and U266/LR6. The results showed that increased expression of FANC/BRCA pathway genes correlated with the melphalan resistance. The formation of ICL at 5 hours after a 2 hour melphalan exposure was reduced in the LR5 compared to the drug sensitive cell 8226 using single cell comet assay. Using siRNA to knock down FANCL or FANCF in melphalan-resistant cell line LR5 reversed the drug resistance. Conversely, overexpression of FANCL or FANCF in the 8226/S drug sensitive cell line enhanced cell survival. These data show that enhanced DNA repair via Fanconi anemia/BRCA pathway is involved in melphalan-resistant myeloma cells.

Milatuzumab Improves Efficacy of Current Treatments for B-Cell Malignancies.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2837-2837
Author(s):  
Rhona Stein ◽  
Susan Chen ◽  
Francisco J. Hernandez-Ilizaliturri ◽  
Myron S. Czuczman ◽  
David M. Goldenberg
Keyword(s):  
Multiple Myeloma ◽  
B Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Proliferative Activity ◽  
Sensitive Cell ◽  
Sensitive Cell Line ◽  
B Cell Malignancies ◽  
Inhibition Of Proliferation

Abstract Abstract 2837 Introduction: Milatuzumab (Immunomedics, Inc.) is a humanized anti-CD74 monoclonal antibody in clinical evaluation for therapy of multiple myeloma, CLL, and NHL. CD74, the MHC class-II chaperone molecule, also functions as the cellular receptor for the proinflammatory cytokine, macrophage migration-inhibitory factor, and initiates a signaling cascade resulting in proliferation and survival. Preclinically, milatuzumab demonstrates therapeutic activity against various B-cell malignancies when used alone, and the therapeutic efficacies of bortezomib, doxorubicin, and dexamethasone are enhanced in multiple myeloma cell lines when given combined with milatuzumab. In addition, milatuzumab acts through distinct mechanisms from rituximab, and exhibits different expression and sensitivity profiles. Here we examine milatuzumab given in combination with rituximab or fludarabine in human NHL, CLL, and ALL cell lines. Methods: Three human NHL (WSU-FSCCL, Raji, and RL); two ALL (MN60 and REH), and two CLL (MEC-1 and WAC) cell lines were tested, with evaluation of therapeutic efficacies of milatuzumab and fludarabine performed in NHL and CLL cell lines. Results: Anti-proliferative activity was augmented in vitro when milatuzumab and rituximab were combined. For example in WSU-FSCCL cells, which are relatively insensitive to rituximab, inhibition of proliferation in the presence of 33.3 nM rituximab increased from 12.6±3.7% in the absence of milatuzumab to 85.5±0.0% (P=.023) in the presence of 33.3 nM milatuzumab. In Raji, a more sensitive cell line, inhibition of proliferation in the presence of 22.2 nM rituximab increased from 64.8±1.3% without milatuzumab to 86.6±0.9% (P=.018) with 22.2 nM milatuzumab. Significant increases in the anti-proliferative activity of rituximab were similarly observed in all but one of the tested NHL, CCL, and ALL cell lines, REH, which was not sensitive to killing by either milatuzumab or rituximab. Unlike rituximab, milatuzumab induces little or no ADCC or CDC. However, in vitro exposure of cells to milatuzumab does not affect rituximab mediated ADCC or CDC. Moreover, the combination of milatuzumab and rituximab was shown to result in a more potent decrease in the mitochondrial potential in rituximab-sensitive cell lines. In the 3 NHL and 2 CLL cell lines, it was found that milatuzumab increased the efficacy of fludarabine. For example, in Raji cells, which are relatively insensitive to fludarabine, inhibition of proliferation in the presence of 4 nM fludarabine increased from no inhibition in the absence of milatuzumab to 76.9±0.7% (P=.009) in the presence of 33.3 nM milatuzumab. In WSU-FSCCL cells, a more fludarabine-sensitive cell line, inhibition of proliferation in the presence of 0.8 nM fludarabine increased from 41.3±0.3% in the absence of milatuzumab to 79.7±0.1% (P<.0001) with 33.3 nM milatuzumab. Conclusions: Milatuzumab, a promising new therapeutic for B-cell malignancies as a naked antibody, can significantly add to the efficacy of currently approved therapies for these diseases, including fludarabine and rituximab. (Supported in part by USPHS grants P01-CA103985 and R01-CA109474 from the NIH and NJDHSS grant 07-1824-FS-N-0.) Disclosures: Goldenberg: Immunomedics, Inc.: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties.

scholarly journals Polymorphism of human minor histocompatibility antigens: T cell recognition of human minor histocompatibility peptides presented by HLA-B35 subtype molecules.

1995 ◽  
Vol 181 (6) ◽  
pp. 2037-2048 ◽  
Author(s):  
Y Beck ◽  
L Satz ◽  
Y Takamiya ◽  
S Nakayama ◽  
L Ling ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Sensitive Cell ◽  
Sensitive Cell Line ◽  
Histocompatibility Antigens ◽  
Minor Histocompatibility Antigens ◽  
T Cell Recognition ◽  
Stimulation Of

To investigate the polymorphism of human minor histocompatibility (mH) antigens, PBLs from 23 Japanese individuals and 25 German individuals with HLA-B35 were studied by using four human mH antigen-specific, HLA-B35-restricted CTL clones. The CTL clones killed PHA-stimulated PBLs from all 23 Japanese individuals. On the other hand, they killed the PHA-stimulated PBLs from 19 of 25 German individuals and partially killed the PHA-stimulated PBLs from three German individuals (CTL weakly sensitive cell line); those from another three individuals (CTL-resistant cell line) were not killed by the CTL clones. All of three CTL weakly sensitive cell lines carry HLA-B*3503 molecules, whereas the three CTL-resistant cell lines carry HLA-B*3502, B*3507, and B*3508 molecules. The cytotoxicity of the CTL clones for three CTL weakly sensitive cell lines was enhanced by stimulation of human mH peptides isolated from HLA-B*3501 molecules purified from C1R-B*3501 cells. Small amounts of human mH peptides were isolated from B*3503 molecules purified from these three CTL weakly sensitive cell lines. Taken together, these results indicate that weak recognition by the CTL clones of three CTL weakly sensitive cell line results from a small amount of the human mH peptides presented by B*3503 molecules. The CTL-resistant cell line carrying B*3507 loaded with the human mH peptides was killed by four CTL clones, whereas the cell lines carrying B*3502 or B*3508 loaded with the peptides were not. The human mH peptides were not isolated from B*3507 molecules purified from the cell lines expressing this subtype, whereas small amounts of the human mH peptides were isolated from B*3502 and B*3508 molecules purified from the cell lines expressing the subtypes. These results indicate that failure of the CTL recognition of the cell line carrying B*3507 is due to a lack of human mH antigens in this cell line. The failure of the CTL recognition of the cell lines carrying B*3502 and B*3508 is not explained by only the amount of the human mH peptides binding to these B35 subtype molecules because the amount of the human mH peptides eluted from B*3502 and B*3508 molecules purified from the cell lines carrying these B35 subtypes is almost the same as that eluted from B*3503 molecules purified from the cell lines carrying B*3503.(ABSTRACT TRUNCATED AT 400 WORDS)

Antioxidant effects and in vitro cytotoxicity on human cancer cell lines of flavonoid-rich Flamboyant (Delonix regia (Bojer) Raf.) flower extract

2020 ◽  
Vol 21 ◽  
Author(s):  
Putthiporn Khongkaew ◽  
Phanphen Wattanaarsakit ◽  
Konstantinos I. Papadopoulos ◽  
Watcharaphong Chaemsawang
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Flower Extract ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Murine Leukemia Cell ◽  
Dose Dependent

Background: Cancer is a noncommunicable disease with increasing incidence and mortality rates both worldwide and in Thailand. Its apparent lack of effective treatments is posing challenging public health issues. Introduction: Encouraging research results indicating probable anti-cancer properties of the Delonix regia flower extract (DRE) have prompted us to evaluate the feasibility of developing a type of product for future cancer prevention or treatment. Methods and Results: In the present report, using High Performance Liquid Chromatography (HPLC), we demonstrate in the DRE, the presence of high concentrations of three identifiable flavonoids, namely rutin 4.15±0.30 % w/w, isoquercitrin 3.04±0.02 %w/w, and myricetin 2.61±0.01 % w/w respectively while the IC50 of DPPH and ABTS assay antioxidation activity was 66.88±6.30 µg/ml and 53.65±7.24 µg/ml respectively. Discussion: Our cancer cell line studies using the MTT assay demonstrated DREs potent and dose dependent inhibition of murine leukemia cell line (P-388: 35.28±4.07% of cell viability remaining), as well as of human breast adenocarcinoma (MCF-7), human cervical carcinoma (HeLa), human oral cavity carcinoma (KB), and human colon carcinoma (HT-29) cell lines in that order of magnitude. Conclusion: Three identifiable flavonoids (rutin, isoquercitrin and myricetin) with high antioxidation activity and potent and dose dependent inhibition of murine leukemia cell line and five other cancer cell lines were documented in the DRE. The extract’s lack of cytotoxicity in 3 normal cell lines is a rare advantage not usually seen in current antineoplastic agents. Yet another challenge of the DRE was its low dissolution rate and long-term storage stability, issues to be resolved before a future product can be formulated.

scholarly journals Phytochemical Analysis and In Vitro Effects of Allium fistulosum L. and Allium sativum L. Extracts on Human Normal and Tumor Cell Lines: A Comparative Study

Molecules ◽  
2021 ◽  
Vol 26 (3) ◽  
pp. 574
Author(s):  
Adrian Bogdan Țigu ◽  
Cristian Silviu Moldovan ◽  
Vlad-Alexandru Toma ◽  
Anca Daniela Farcaș ◽  
Augustin Cătălin Moț ◽  
...  
Keyword(s):  
Cell Line ◽  
Allium Sativum ◽  
Allium Fistulosum ◽  
Tumor Cell Lines ◽  
Sensitive Cell ◽  
Sensitive Cell Line ◽  
Welsh Onion ◽  
Normal Cells ◽  
Ic50 Value ◽  
High Concentrations

Allium sativum L. (garlic bulbs) and Allium fistulosum L. (Welsh onion leaves) showed quantitative differences of identified compounds: allicin and alliin (380 µg/mL and 1410 µg/mL in garlic; 20 µg/mL and 145 µg/mL in Welsh onion), and the phenolic compounds (chlorogenic acid, p-coumaric acid, ferulic acid, gentisic acid, 4-hydroxybenzoic acid, kaempferol, isoquercitrin, quercitrin, quercetin, and rutin). The chemical composition determined the inhibitory activity of Allium extracts in a dose-dependent manner, on human normal cells (BJ-IC50 0.8841% garlic/0.2433% Welsh onion and HaCaT-IC50 1.086% garlic/0.6197% Welsh onion) and tumor cells (DLD-1-IC50 5.482%/2.124%; MDA-MB-231-IC50 6.375%/2.464%; MCF-7-IC50 6.131%/3.353%; and SK-MES-1-IC50 4.651%/5.819%). At high concentrations, the cytotoxic activity of each extract, on normal cells, was confirmed by: the 50% of the growth inhibition concentration (IC50) value, the cell death induced by necrosis, and biochemical determination of LDH, catalase, and Caspase-3. The four tumor cell lines treated with high concentrations (10%, 5%, 2.5%, and 1.25%) of garlic extract showed different sensibility, appreciated on the base of IC50 value for the most sensitive cell line (SK-MES-1), and the less sensitive (MDA-MB-231) cell line. The high concentrations of Welsh onion extract (5%, 2.5%, and 1.25%) induced pH changes in the culture medium and SK-MES-1 being the less sensitive cell line.

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