Functional assay of multidrug resistant cells using JC-1, a carbocyanine fluorescent probe

Kühnel, JM; Jy, Perrot; Faussat, Anne Marie; Marie, J; Schwaller, M. A.

doi:10.1038/sj.leu.2400698

Cited by 52 publications

(43 citation statements)

References 3 publications

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“…The dye enters living cells and fluoresces bright red in its multimeric form within active mitochondria. In apoptotic cells, the mitochondrial membrane potential collapses, and the JC-1 dye does not manifest its multimeric form within the mitochondria, thus remaining in the cytoplasm in its green J-monomeric form (Kuhnel et al 1997). We verified that DREF over-expression under the GMR-GAL4 driver induces apoptosis in eye imaginal disc cells (Fig.…”

Section: Dref Is a Key Regulator For Dant Promoter Activity In Livingmentioning

confidence: 54%

Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system

et al. 2007

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Adenine nucleotide translocase (ANT) is a crucial component in the maintenance of cellular energy homeostasis, as well as in the formation of the mitochondrial permeability transition pores. However, the molecular mechanisms regulating the expression of the ANT gene are poorly understood. In this study, we have identified three DNA replication-related elements (DRE; 5′ ′ ′ ′-TATCGATA) in the 5′ ′ ′ ′-flanking region of the Drosophila ANT (dANT ) gene. Gel-mobility shift analyses revealed that all three of the DREs were recognized by the DRE-binding factor (DREF). The site-directed mutagenesis of these DRE sites induces a considerable reduction in the activity of the dANT gene promoter in vitro. Analyses with transgenic flies harboring a dANT-lacZ fusion gene bearing the wild-type or mutant DRE sites showed that the DRE sites were required for the expression of dANT in vivo. We determined that the over-expression or knockdown of DREF exerts a regulatory effect on the activity of the dANT promoter. In addition, we observed the collapse of mitochondrial membrane potential in the eye imaginal discs in which DREF was overexpressed. These results show that DRE/DREF is a crucial regulator of dANT gene expression, and also suggest the possibility that cross-talk may occur between the DRE/DREF system and mitochondrial functioning.

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Section: Dref Is a Key Regulator For Dant Promoter Activity In Livingmentioning

confidence: 54%

Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system

et al. 2007

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“…However, using K562 cell lines with increasing levels of resistance, we demonstrated that the level of JC-1 accumulation correlated with the level of P-gp expression as detected by UIC2 antibodies (8). Also, in AML patients, JC-1 was shown to be able to discern between three groups of patients: resistant, intermediate, and sensitive (9).…”

Section: Discussionmentioning

confidence: 84%

“…With JC-1, the sensitive cell lines display both green and red fluorescence. In resistant cells, with the increase of P-gp activity the green fluorescence of JC-1 decreased, and the red fluorescence was lost completely (8). Our study thus proved JC-1 to be a very sensitive probe, more sensitive than Rhodamine 123 (Rh 123), for the detection of low-level P-gp activity in adult patients with acute myeloid leukemia (AML).…”

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confidence: 77%

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JC‐1, a sensitive probe for a simultaneous detection of P‐glycoprotein activity and apoptosis in leukemic cells

Chaoui

Faussat

Majdak

et al. 2006

Cytometry Part B Clinical

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Background: JC-1 probe has been successfully used for the analysis of either apoptosis or P-glycoprotein (P-gp) activity. Therefore, we wanted to see if JC-1 could also simultaneously assess both, P-gp activity and apoptosis, in acute myeloid leukemia (AML) cells.Methods Conclusions: JC-1 can simultaneously evaluate two important parameters involved in drug resistance in AML cells, P-gp activity and apoptosis. q

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“…32 Verapamil pretreatment did not affect fluorescence in LoVo WT cells, while the emitted fluorescence was significantly increased in MDR cells.…”

Section: Flow Cytometrymentioning

confidence: 85%

Enzymatic oxidation products of spermine induce greater cytotoxic effects on human multidrug‐resistant colon carcinoma cells (LoVo) than on their wild‐type counterparts

Calcabrini¹,

Arancia²,

Marra³

et al. 2002

Intl Journal of Cancer

122

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The occurrence of resistance to cytotoxic agents in tumor cells, associated with several phenotypic alterations, is one of the major obstacles to successful anticancer chemotherapy. A new strategy to overcome MDR of human cancer cells was studied, using BSAO, which generates cytotoxic products from spermine, H 2 O 2 and aldehyde(s). The involvement of these products in causing cytotoxicity was investigated in both drug-sensitive (LoVo WT) and drug-resistant (LoVo DX) colon adenocarcinoma cells. Evaluation of clonogenic cell survival showed that LoVo DX cells are more sensitive than LoVo WT cells. Fluorometric assay and treatments performed in the presence of catalase demonstrated that the cytotoxicity was due mainly to the presence of H 2 O 2 . Cytotoxicity was eliminated in the presence of both catalase and ALDH. Transmission electron microscopic observations showed more pronounced mitochondrial modifications in drug-resistant than in drug-sensitive cells. Mitochondrial functionality studies performed by flow cytometry after JC-1 labeling revealed basal hyperpolarization of the mitochondrial membrane in LoVo DX cells. After treatment with BSAO and spermine, earlier and higher mitochondrial membrane depolarization was found in LoVo DX cells than in drug-sensitive cells. In addition, higher basal ROS production in LoVo DX cells than in drug-sensitive cells was detected by flow-cytometric analysis, suggesting increased mitochondrial activity in drug-resistant cells. Our results support the hypothesis that mitochondrial functionality affects the sensitivity of cells to the cytotoxic enzymatic oxidation products of spermine, which might be promising anticancer agents, mainly against drug-resistant tumor cells.

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Functional assay of multidrug resistant cells using JC-1, a carbocyanine fluorescent probe

Cited by 52 publications

References 3 publications

Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system

Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system

JC‐1, a sensitive probe for a simultaneous detection of P‐glycoprotein activity and apoptosis in leukemic cells

Enzymatic oxidation products of spermine induce greater cytotoxic effects on human multidrug‐resistant colon carcinoma cells (LoVo) than on their wild‐type counterparts

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