Expression data from multiple myeloma cells treated with arsenic
ABSTRACT: We used microarrays to examine changes in gene expression in multiple myeloma cell lines following treatment with arsenic trioxide and darinaparsin Keywords: Time course Overall design: Four multiple myeloma cell lines (U266, MM.1s, KMS11, 8226/S) were treated with either arsenic trioxide (ATO) for 6, 24, or 48 hours or darinaparsin (DAR) for 6 or 24 hours; RNA was extracted from treated and control cells for microarray analysis
Project description:We used microarrays to examine changes in gene expression in multiple myeloma cell lines following treatment with arsenic trioxide and darinaparsin Experiment Overall Design: Four multiple myeloma cell lines (U266, MM.1s, KMS11, 8226/S) were treated with either arsenic trioxide (ATO) for 6, 24, or 48 hours or darinaparsin (DAR) for 6 or 24 hours; RNA was extracted from treated and control cells for microarray analysis
Project description:The aim of this study was to gain insight into the potential mechanism of resistance to arsenic trioxide (ATO). The gene expression profile of naive (NB4) (Acute promyelocytic leukemia (APL) cell line and one of its in house generated ATO resistant sub clone (NB4-VM-AsR1) was done using whole genome microarray and compared to generate the differential expression profile which will give insight into the mechanisms of ATO resistance in APL. Agilent one-color experiment,Organism: Human ,Agilent Whole Genome Human 4x44k (AMADID: 014850) , Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442) naive versus arsenic trioxide resistant acute promyelocytic leukemia cell line NB4
Project description:We performed genome-wide DNA methylation profiling of KMS11, MM.1S, and RPMI8226 multiple myeloma cell lines to identify methylation changes distinct to each cell line Three multiple myeloma cell lines (KMS11, MM.1S, and RPMI8226) with two replicates each
Project description:We performed Illumina Infinium whole-genome SNP-CN profiling of KMS11, MM.1S, and RPMI8226 multiple myeloma cell lines to detect gene copy number variants distinct to each cell line Three multiple myeloma cell lines (KMS11, MM.1S, and RPMI8226)
Project description:Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of malignant CD5+ B lymphocytes (CLL cells) in the peripheral blood, and their progressive infiltration in lymphoid organs. Despite the efforts made, CLL remains an incurable disease. It is therefore crucial to continue searching for new therapeutic agents and targets. Arsenic trioxide (ATO) induces apoptosis in all CLL cases and it could constitute an efficient therapy for this disease. We used Affimetrix microarray technology to characterize the global gene expression profile underlying CLL cell response to ATO, with the aim of identifying the ATO mechanism of action as well as new therapeutic targets in CLL. Overall design: MEC-1 cells (CLL-derived) were incubated with or without 5 µM ATO for 24 h and total RNA was extracted and used to hybridize an Affymetrix GeneChip® Human Gene 1.0 ST Array.
Project description:The purpose of this study is to search for aberrant genes in HaCaT keratinocytes after chronic exposure to arsenic trioxide. The objective of the investigation was to discover the mechanism of arsenic carcinogenicity in human epidermal keratinocytes. We hypothesize that a combined strategy of DNA microarray, qRT-PCR and gene function annotation will identify aberrantly expressed genes in HaCaT keratinocyte cell line after chronic treatment with arsenic trioxide. HaCaT cells were chronically exposed to 0.5µg/mL arsenic trioxide (As2O3) up to 22 passages and RNA was extracted. Microarray data analysis identified 14 up-regulated genes and 21 down-regulated genes in response to arsenic trioxide Two experimental groups: 1. The treatment group was sub-cultured up to passage 22 to establish a chronic exposure state. 2. The passage control group was also sub-cultured up to 22 passages but with no exposure to arsenic trioxide. 4 technical replicates with 3 replicates making a total of 8X3 =24 samples HaCat Cell untreated (passage control): 1. H1_H001, H1_H002, H1_H003 2. H2_ H004, H2_H005, H2_H006 3. H3_ H007, H3_H008, H3_H009 4. H4_ H010, H4_H011, H4_H012 HaCat Cell treated with 0.5µg/ml of arsenic trioxide: 5. A1_H013, A1_H014, A1_H015 6. A2_H016, A2_H017, A2_H018 7. A3_H019, A3_H020, A3_H021 8. A4_H022, A4_H023, A4_H024 Cell Type: Human Skin Keratinocyte: 1.5 ×105 HaCaT cells were cultured in 7.5 ml of complete DMEM containing 10% Fetal Bovine Serum (FBS) and 1% penicillin, streptomycin in T-25 culture plate. Cells were incubated in a humidified atmosphere with 5% CO2 at 37 ºC. The treatment groups were exposed to 0.5µg/mL As2O3 (equivalent to LC 0.5), and passaged at 90% confluent. Total RNA was extracted from 4 technical replicates of unexposed HaCaT cells and HaCaT cells chronically exposed to arsenic trioxide up to passage 22 using RNA STAT-60 (TEL-TEST, INC, Friendswood, TX, USA).
Project description:Arsenic trioxide (ATO) and all-trans retinoic acid (ATRA) combination safely cures fatal acute promyelocytic leukemia, but the mechanisms underlying their action and synergy remain elusive. ATRA inhibits APL, breast and liver cancers by targeting isomerase Pin1, a master regulator of oncogenic signaling. Here we show that ATO targets Pin1 and cooperates with ATRA to exert potent anticancer activity. ATO inhibits and degrades Pin1, and suppresses its oncogenic function by noncovalent binding to Pin1’s active site. ATRA increases cellular ATO uptake through upregulating aquaporin-9. ATO and ATRA, at clinically safe doses, cooperatively ablate Pin1 to block numerous cancer-driving pathways and inhibit the growth of triple-negative breast cancer cells and tumor-initiating cells in cell and animal models including patient-derived orthotopic xenografts, similar to Pin1 CRISPR knockout, which is substantiated by comprehensive protein and microRNA analyses. Thus, synergistic Pin1 inhibition by ATO and ATRA offers an attractive approach to combating breast and other cancers.
Project description:Darinaparsin (DPS) is an arsenic cytotoxin with a more favorable toxicity profile than the established anti-leukemic drug, arsenic trioxide (ATO). Here we report effects of DPS on a variety of solid tumor cell lines under normoxia as well as hypoxia, the latter an important characteristic of the tumor microenvironment. Using MTT and clonogenic assays, we demonstrated that DPS had potent cytotoxic activities under both normoxia and hypoxia, with IC50??s ~ 2- to 3-fold lower than ATO. Xenograft studies using tumor cell lines as well as patient-derived tumor tissues implanted under the renal capsule in mice confirmed the anti-tumor activity of DPS in vivo. DPS was also a more potent radiosensitizer under hypoxia than ATO in vitro, and sensitized solid tumors to radiation in vivo at doses that had no systemic toxicities. Interestingly, in contrast to previous reports of DPS effects on leukemic cells under normoxia, DPS-induced killing of hypoxic solid tumor cells was not dependent on ROS generation and oxidative damage. Instead, cDNA microarray analysis suggested that DPS inhibited oncogene- (such as RAS and MYC) associated gene expression. In fact, compared to normal mouse embryonic fibroblasts (MEFs), oncogene (RAS/E1A) transformed MEFs were markedly more sensitive to DPS-induced apoptosis under both normoxia and hypoxia. Altogether, these results demonstrate that DPS has significant and preferential cytotoxicity against solid tumor as compared with normal cells, and is an effective radiosensitizer. Since DPS is in early clinical development as a single agent, these findings have near term translational potential. time_series_design
Project description:Gene expression (GE) profiling of multiple myeloma (MM) cells is a promising means of identifying high-risk MM patients. The analyses depend on plasma cell purification by CD138+ cell separation. Considering the sensitivity of gene transcription, we wanted to test if cell separation distorts true in vivo GE patterns. We performed a controlled study of running 4 human myeloma cell lines (HMCLs: U266, INA-6, RPMI 8226, and NCI H929) through a CD138+ separation procedure identical to the handling of clinical samples. We then compared the resulting effects on gene expression. Using U266 as a screening model, we performed global GE analysis using the Affymetrix Human Gene 1.0 ST array. Sample cells showed significant changes (adj. p<0,05) in 670 genes compared to the non-separated controls. We searched for upregulated genes of myeloma and/or cancer relevance and chose (in decending fold change order) FOS, DUSP1, MIRN21, NFKBIA, and ATF4 for PCR validation. Note: Only U266 cells were used for microarray analysis. The other cell lines were used later to validate the array results.
Project description:Aberrations in the Hedgehog (Hh) pathway are known to related to several malignancies. However, little is known about the function of GLI2, a transcription factor in the Hh pathway, in osteosarcoma. Osteosarcoma is the most frequent primary bone sarcoma in children and adolescents. Despite survival rates of osteosarcoma patients have increased, the prognosis of patients with metastasis remains poor. Therefore, the development of novel therapeutic strategies for osteosarcoma patients is development of novel therapeutic strategies for osteosarcoma patients is urgently needed. Aberrations in the Hedgehog (Hh) pathway are known to related to several malignancies. However, little is known about the function of GLI2, a transcription factor in the Hh pathway, in osteosarcoma. Our findings revealed that GLI2 was overexpressed in osteosarcoma tissues. Additionally, GLI2 is involved in the metastasis of osteosarcoma cells through the regulation of ribosomal protein S3 expression. Furthermore, we showed that arsenic trioxide (ATO) suppressed the invasion and lung metastasis of osteosarcoma cells by the inhibition of GLI transcription. Consequently, these finding reveal a novel function of GLI2 in the metastasis of osteosarcoma and that ATO may be a new therapeutic agentay be a new therapeutic agent. We revealed that a novel function of GLI2 in the metastasis of osteosarcoma and that ATO may be a new therapeutic agent for preventing osteosarcoma metastasis. Negative siRNA(U-2OS) and GLI2 siRNA(U-2OS)