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Inhibition of malignant phenotypes of human osteosarcoma cells by a gene silencer, a pyrrole-imidazole polyamide, which targets an E-box motif.

Taniguchi M, Fujiwara K, Nakai Y, Ozaki T, Koshikawa N, Toshio K, Kataba M, Oguni A, Matsuda H, Yoshida Y, Tokuhashi Y, Fukuda N, Ueno T, Soma M, Nagase H - FEBS Open Bio (2014)

Bottom Line: Gene amplification and/or overexpression of the transcription factor c-MYC, which binds to the E-box sequence (5'-CACGTG-3'), has been observed in many human tumors.In this study, we have designed 5 pyrrole-imidazole (PI) polyamides recognizing E-box, and found that, among them, Myc-6 significantly suppresses malignant phenotypes of human osteosarcoma MG63 cells both in vitro and in vivo.Collectively, our present findings strongly suggest that Myc-6 exerts its tumor-suppressive ability at least in part through the specific down-regulation of MALAT1.

View Article: PubMed Central - PubMed

Affiliation: Division of Orthopedic Surgery, Nihon University School of Medicine, 30-1 Oyaguchi Kami-Cho, Itabashi, Tokyo 173-8610, Japan.

ABSTRACT
Gene amplification and/or overexpression of the transcription factor c-MYC, which binds to the E-box sequence (5'-CACGTG-3'), has been observed in many human tumors. In this study, we have designed 5 pyrrole-imidazole (PI) polyamides recognizing E-box, and found that, among them, Myc-6 significantly suppresses malignant phenotypes of human osteosarcoma MG63 cells both in vitro and in vivo. Intriguingly, knockdown of the putative Myc-6 target MALAT1 encoding long noncoding RNA remarkably impaired cell growth of MG63 cells. Collectively, our present findings strongly suggest that Myc-6 exerts its tumor-suppressive ability at least in part through the specific down-regulation of MALAT1.

No MeSH data available.


Related in: MedlinePlus

Myc-6 significantly represses cell viability of human osteosarcoma MG63 cells. (A) Sequence comparison between the putative targets of the indicated synthetic PI polyamides and the consensus E-box. Sequences corresponding to E-box are underlined. W indicates A or T. (B) WST-8 assay. MG63 cells were exposed to water (control), non-specific PI polyamide (mismatch) or with the indicated PI polyamides. Six days after the treatment, cell viability was examined by WST-8 assay. Data are expressed as the means ± SD. Differences were considered significant at p < 0.05. ∗p < 0.01;∗∗p < 0.001. (C) Chemical structure of Myc-6. (D) Schematic representation of DNA recognition by Myc-6. Open circle, filled circle, β and Dp indicate pyrrole, imidazole, β-alanine and dimethylpropanediamine, respectively. (E) Dose and time-dependency. Cell viability was measured by WST8 assay 3–6 days after addition of Myc-6. Differences between viability of cells treated with or without Myc-6 were considered significant at p < 0.05. ∗p < 0.05, ∗∗p < 0.01. Data were expressed as the means ± SD.
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f0005: Myc-6 significantly represses cell viability of human osteosarcoma MG63 cells. (A) Sequence comparison between the putative targets of the indicated synthetic PI polyamides and the consensus E-box. Sequences corresponding to E-box are underlined. W indicates A or T. (B) WST-8 assay. MG63 cells were exposed to water (control), non-specific PI polyamide (mismatch) or with the indicated PI polyamides. Six days after the treatment, cell viability was examined by WST-8 assay. Data are expressed as the means ± SD. Differences were considered significant at p < 0.05. ∗p < 0.01;∗∗p < 0.001. (C) Chemical structure of Myc-6. (D) Schematic representation of DNA recognition by Myc-6. Open circle, filled circle, β and Dp indicate pyrrole, imidazole, β-alanine and dimethylpropanediamine, respectively. (E) Dose and time-dependency. Cell viability was measured by WST8 assay 3–6 days after addition of Myc-6. Differences between viability of cells treated with or without Myc-6 were considered significant at p < 0.05. ∗p < 0.05, ∗∗p < 0.01. Data were expressed as the means ± SD.

Mentions: PI polyamides targeting E-box were designed as shown in Fig. 1A and synthesized as described previously [16]. All of the PI polyamides except for Myc-3 were dissolved in distilled water (at a final concentration of 1 mM) and small aliquots were stored at −20 °C. Myc-3 was dissolved in 50% DMSO and small aliquots were kept at −20 °C.


Inhibition of malignant phenotypes of human osteosarcoma cells by a gene silencer, a pyrrole-imidazole polyamide, which targets an E-box motif.

Taniguchi M, Fujiwara K, Nakai Y, Ozaki T, Koshikawa N, Toshio K, Kataba M, Oguni A, Matsuda H, Yoshida Y, Tokuhashi Y, Fukuda N, Ueno T, Soma M, Nagase H - FEBS Open Bio (2014)

Myc-6 significantly represses cell viability of human osteosarcoma MG63 cells. (A) Sequence comparison between the putative targets of the indicated synthetic PI polyamides and the consensus E-box. Sequences corresponding to E-box are underlined. W indicates A or T. (B) WST-8 assay. MG63 cells were exposed to water (control), non-specific PI polyamide (mismatch) or with the indicated PI polyamides. Six days after the treatment, cell viability was examined by WST-8 assay. Data are expressed as the means ± SD. Differences were considered significant at p < 0.05. ∗p < 0.01;∗∗p < 0.001. (C) Chemical structure of Myc-6. (D) Schematic representation of DNA recognition by Myc-6. Open circle, filled circle, β and Dp indicate pyrrole, imidazole, β-alanine and dimethylpropanediamine, respectively. (E) Dose and time-dependency. Cell viability was measured by WST8 assay 3–6 days after addition of Myc-6. Differences between viability of cells treated with or without Myc-6 were considered significant at p < 0.05. ∗p < 0.05, ∗∗p < 0.01. Data were expressed as the means ± SD.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4048845&req=5

f0005: Myc-6 significantly represses cell viability of human osteosarcoma MG63 cells. (A) Sequence comparison between the putative targets of the indicated synthetic PI polyamides and the consensus E-box. Sequences corresponding to E-box are underlined. W indicates A or T. (B) WST-8 assay. MG63 cells were exposed to water (control), non-specific PI polyamide (mismatch) or with the indicated PI polyamides. Six days after the treatment, cell viability was examined by WST-8 assay. Data are expressed as the means ± SD. Differences were considered significant at p < 0.05. ∗p < 0.01;∗∗p < 0.001. (C) Chemical structure of Myc-6. (D) Schematic representation of DNA recognition by Myc-6. Open circle, filled circle, β and Dp indicate pyrrole, imidazole, β-alanine and dimethylpropanediamine, respectively. (E) Dose and time-dependency. Cell viability was measured by WST8 assay 3–6 days after addition of Myc-6. Differences between viability of cells treated with or without Myc-6 were considered significant at p < 0.05. ∗p < 0.05, ∗∗p < 0.01. Data were expressed as the means ± SD.
Mentions: PI polyamides targeting E-box were designed as shown in Fig. 1A and synthesized as described previously [16]. All of the PI polyamides except for Myc-3 were dissolved in distilled water (at a final concentration of 1 mM) and small aliquots were stored at −20 °C. Myc-3 was dissolved in 50% DMSO and small aliquots were kept at −20 °C.

Bottom Line: Gene amplification and/or overexpression of the transcription factor c-MYC, which binds to the E-box sequence (5'-CACGTG-3'), has been observed in many human tumors.In this study, we have designed 5 pyrrole-imidazole (PI) polyamides recognizing E-box, and found that, among them, Myc-6 significantly suppresses malignant phenotypes of human osteosarcoma MG63 cells both in vitro and in vivo.Collectively, our present findings strongly suggest that Myc-6 exerts its tumor-suppressive ability at least in part through the specific down-regulation of MALAT1.

View Article: PubMed Central - PubMed

Affiliation: Division of Orthopedic Surgery, Nihon University School of Medicine, 30-1 Oyaguchi Kami-Cho, Itabashi, Tokyo 173-8610, Japan.

ABSTRACT
Gene amplification and/or overexpression of the transcription factor c-MYC, which binds to the E-box sequence (5'-CACGTG-3'), has been observed in many human tumors. In this study, we have designed 5 pyrrole-imidazole (PI) polyamides recognizing E-box, and found that, among them, Myc-6 significantly suppresses malignant phenotypes of human osteosarcoma MG63 cells both in vitro and in vivo. Intriguingly, knockdown of the putative Myc-6 target MALAT1 encoding long noncoding RNA remarkably impaired cell growth of MG63 cells. Collectively, our present findings strongly suggest that Myc-6 exerts its tumor-suppressive ability at least in part through the specific down-regulation of MALAT1.

No MeSH data available.


Related in: MedlinePlus