Inhibition of P-Glycoprotein and Recovery of Drug Sensitivity of Human Acute Leukemic Blast Cells by Multidrug Resistance Gene (mdr1) Antisense Oligonucleotides

Motomura, Sayuri; Motoji, Toshiko; Takanashi, Minoko; Wang, Yanhua; Shiozaki, Hiroko; Sugawara, Isamu; Aikawa, Eizou; Tomida, Akihiro; Tsuruo, Takashi; Kanda, Naotoshi; Mizoguchi, Hideaki

doi:10.1182/blood.v91.9.3163

Cited by 45 publications

(14 citation statements)

References 32 publications

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“…The P-gp was induced in these cells by a single-step selection after exposure to low DOX concentrations equivalent to that found in leukemia patients. 29 Unlike engineered cell lines with higher surface concentrations of P-gp, the native-like cells used in that study might have been more responsive to silencing due to lower concentrations of the transporters. However, even engineered cells with high P-gp levels were reported to respond well to shRNAmediated silencing; for example, sensitivity of human colon carcinoma HCT-8 cells displayed increased sensitivity as IC 50 decreased (based on IC 50 assessment after vincristine treatment) 66 or multidrug-resistant gastric carcinoma EPG85-257RDB cells reverted completely to drugsensitive phenotype (with daunorubicin) after stable shRNA expression.…”

Section: Expression Of P-gp Silencers In Situmentioning

confidence: 92%

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Recent attempts at RNAi‐mediated P‐glycoprotein downregulation for reversal of multidrug resistance in cancer

Abbasi

Lavasanifar

Uludağ

2011

Medicinal Research Reviews

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Multidrug resistance (MDR) is among the major mechanisms leading to failure in chemotherapy of cancer patients. The ATP-binding cassette proteins are major contributors to MDR, involved in the active efflux of xenobiotics out of cancer cells. Among them, P-glycoprotein (P-gp) is the most dominant protein involved in the efflux of drugs. For more than 30 years, scientists have searched for the ideal P-gp inhibitor to modulate drug resistance activity of P-gp. This inhibitor should be tissue and cell specific with side effects on other tissues, must not provoke immune responses from the host, should provide sustained inhibition, and must be synthesized readily with low cost. Chemical P-gp inhibitors tested to date, have shown nonspecific toxic effects limiting their clinical applications. Sequence-specific P-gp gene silencing by RNA interference (RNAi) may provide a more effective approach for downregulation of specific protein targets due to high specificity, limited toxicity and immunogenicity, and relative ease in synthesis. RNAi can be implemented by delivery of synthetic small interfering RNAs (siRNAs) or by gene expression of short hairpin RNAs using gene expressing vectors. Specific delivery systems and expression vectors have been designed for this purpose and many researchers have explored their effectiveness for P-gp downregulation. In this report, we review the efficiency of various methods for siRNA delivery and transfection for P-gp downregulation in cancer cells for MDR reversal. Novel ideas and observations by different research groups were discussed for future improvement in this essential field.

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Section: Expression Of P-gp Silencers In Situmentioning

confidence: 92%

“…Motomura et al used a cationic liposome (Lypolymine) to deliver ASOs to K562 myelogenous leukemia cells and its adriamycin-resistant phenotype K562/ ADM; 16 25-75% P-gp downregulation was achieved at 2.5-20 mM ASO concentrations. 29…”

Section: Specific Downregulation Of P-gp By Rna Interferencementioning

confidence: 99%

Recent attempts at RNAi‐mediated P‐glycoprotein downregulation for reversal of multidrug resistance in cancer

Abbasi

Lavasanifar

Uludağ

2011

Medicinal Research Reviews

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“…Inhibition of Pgp-mediated drug extrusion may allow chemosensitivity of cancer cells to antineoplastic drugs and result in successful treatment of MDR cells. Antisense oligonucleotides (ASODN) and hammerhead ribozymes for specific inhibition of Pgp expression in some malignant tumours have been well established [7,8]. However, ribozymes are RNA molecules that are unstable in cell medium and are easily degradable [9], thus making them inconvenient for experimental use.…”

Section: Introductionmentioning

confidence: 99%

Screening of deoxyribozyme with high reversal efficiency against multidrug resistance in breast carcinoma cells

Gao

Wei

et al. 2011

J Cellular Molecular Medi

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Specific inhibition of P-glycoprotein (Pgp) expression, which is encoded by multidrug resistance gene-1 (MDR1), is considered a well-respected strategy to overcome multidrug resistance (MDR). Deoxyribozymes (DRz) are catalytic nucleic acids that could cleave a target RNA in sequence-specific manner. However, it is difficult to select an effective target site for DRz in living cells. In this study, target sites of DRz were screened according to MDR1 mRNA secondary structure by RNA structure analysis software. Twelve target sites on the surface of MDR1 mRNA were selected. Accordingly, 12 DRzs were synthesized and their suppression effect on the MDR phenotype in breast cancer cells was confirmed. The results showed that 4 (DRz 2, 3, 4, 9) of the 12 DRzs could, in a dose-dependent response, significantly suppress MDR1 mRNA expression and restore chemosensitivity in breast cancer cells with MDR phenotype. This was especially true of DRz 3, which targets the 141 site purine-pyrimidine dinucleotide. Compared with antisense oligonucleotide or anti-miR-27a inhibitor, DRz 3 was more efficient in suppressing MDR1 mRNA and Pgp protein expression or inhibiting Pgp function. The chemosensitivity assay also proved DRz 3 to be the best one to reverse the MDR phenotype. The present study suggests that screening targets of DRzs according to MDR1 mRNA secondary structure could be a useful method to obtain workable ones. We provide evidence that DRzs (DRz 2, 3, 4, 9) are highly efficient at reversing the MDR phenotype in breast carcinoma cells and restoring chemosensitivity.

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“…Numerous attempts have been made to overcome cancer cell resistance against antineoplastic drugs including ADM by inhibition of intracellular resistance factors. 1,2) We previously have reported that endogenous tumor necrosis factor (enTNF) exerts a protective effect against ADM by enhancing the scavenging of intracellular reactive oxygen species (ROS) via induction of manganous superoxide dismutase (MnSOD). 3) We also found that measurement of enTNF expression could predict ADM sensitivity in tumor cells.…”

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confidence: 99%

Augmented Adriamycin Sensitivity in Cells Transduced with an Antisense Tumor Necrosis Factor Gene Is Mediated by Caspase‐3 Downstream from Reactive Oxygen Species

Sasaki

Kobayashi

Watanabe

2001

Japanese Journal of Cancer Research

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While transduction of an antisense tumor necrosis factor (TNF) gene sequence can augment the cytotoxicity of adriamycin (ADM) in human cancer cells, the specific effect of introducing this sequence on the signal transduction pathway leading to cell death remains unclear. In ADM-resistant pancreatic carcinoma (PANC-1) cells, both the antioxidant N-acetyl-L-cysteine (NAC) and the caspase-3 inhibitor acetyl-L-aspartyl-L-methionyl-L-glutaminyl-L-aspartyl-aldehyde (Ac-DMQD-CHO) prevented ADM-induced cytotoxicity. NAC additionally inhibited caspase-3 activity induced by ADM treatment, while Ac-DMQD-CHO showed no suppressive effect on reactive oxygen species (ROS). Stable antisense-TNF transfectants showed higher ADM sensitivity and greater ADMinduced ROS production and caspase-3 activity than mock transfectant or parent cells. These results indicate that increased caspase-3 activity downstream from ROS production is among the mechanisms by which transduction of the antisense TNF sequence of augments ADM sensitivity of pancreatic carcinoma cells.Key words: Tumor necrosis factor -Antisense -Reactive oxygen species -Caspase-3 Adriamycin (ADM) is widely used to treat solid tumors and malignant hematologic diseases. Numerous attempts have been made to overcome cancer cell resistance against antineoplastic drugs including ADM by inhibition of intracellular resistance factors.1, 2) We previously have reported that endogenous tumor necrosis factor (enTNF) exerts a protective effect against ADM by enhancing the scavenging of intracellular reactive oxygen species (ROS) via induction of manganous superoxide dismutase (MnSOD). 3)We also found that measurement of enTNF expression could predict ADM sensitivity in tumor cells. 4) Thus, antisense TNF gene transduction could augment ADMinduced toxicity in human cancer cells by decreasing MnSOD activity. 4,5) However, whether transduction of the antisense TNF gene actually affects the signal transduction pathway leading to ADM-induced cell death is unclear. We therefore investigated changes following antisense TNF gene transduction in expression of molecules leading to cell death in human pancreatic carcinoma (PANC-1) cells showing resistance to ADM. Our investigative group and others recently identified a signaling pathway including production of reactive oxygen species and activation of caspase-3 that leads to apoptosis in cells exposed to heat or ionizing radiation.6, 7) We therefore examined whether and how antisense TNF gene transduction augmented this signaling pathway to result in increased ADM sensitivity. As the PANC-1 cells used in this study harbor a mutant-type p53, effects from several molecules that interact with caspase-3 in cells with wild-type p53 could be excluded. MATERIALS AND METHODSCell culture and gene transduction PANC-1 cells were cultured in RPMI-1640 (Gibco, Grand Island, NY) supplemented with 10% heat-inactivated fetal calf serum (FCS; Biological Industries, Kibbutzbeit Haemek, Israel). Transduction of antisense TNF mRNA expression vector (pLJantiTNF) to PAN...

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Inhibition of P-Glycoprotein and Recovery of Drug Sensitivity of Human Acute Leukemic Blast Cells by Multidrug Resistance Gene (mdr1) Antisense Oligonucleotides

Cited by 45 publications

References 32 publications

Recent attempts at RNAi‐mediated P‐glycoprotein downregulation for reversal of multidrug resistance in cancer

Recent attempts at RNAi‐mediated P‐glycoprotein downregulation for reversal of multidrug resistance in cancer

Screening of deoxyribozyme with high reversal efficiency against multidrug resistance in breast carcinoma cells

Augmented Adriamycin Sensitivity in Cells Transduced with an Antisense Tumor Necrosis Factor Gene Is Mediated by Caspase‐3 Downstream from Reactive Oxygen Species

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