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Epigenetic modulation with HDAC inhibitor CG200745 induces anti-proliferation in non-small cell lung cancer cells.

Chun SM, Lee JY, Choi J, Lee JH, Hwang JJ, Kim CS, Suh YA, Jang SJ - PLoS ONE (2015)

Bottom Line: Histone modification plays a pivotal role on gene regulation, as regarded as global epigenetic markers, especially in tumor related genes.Treatment with CG200745 increased the global level of histone acetylation, resulting in the inhibition of cell proliferation.ChIP-on-chip analysis with an H4K16ac antibody showed altered H4K16 acetylation on genes critical for cell growth inhibition, although decreased at the transcription start site of a subset of genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Asan Medical Center, The University of Ulsan College of Medicine, Seoul, Republic of Korea.

ABSTRACT
Histone modification plays a pivotal role on gene regulation, as regarded as global epigenetic markers, especially in tumor related genes. Hence, chemical approaches targeting histone-modifying enzymes have emerged onto the main stage of anticancer drug discovery. Here, we investigated the therapeutic potentials and mechanistic roles of the recently developed histone deacetylase inhibitor, CG200745, in non-small cell lung cancer cells. Treatment with CG200745 increased the global level of histone acetylation, resulting in the inhibition of cell proliferation. ChIP-on-chip analysis with an H4K16ac antibody showed altered H4K16 acetylation on genes critical for cell growth inhibition, although decreased at the transcription start site of a subset of genes. Altered H4K16ac was associated with changes in mRNA expression of the corresponding genes, which were further validated in quantitative RT-PCR and western blotting assays. Our results demonstrated that CG200745 causes NSCLC cell growth inhibition through epigenetic modification of critical genes in cancer cell survival, providing pivotal clues as a promising chemotherapeutics against lung cancer.

No MeSH data available.


Related in: MedlinePlus

Changes in protein expression of NSCLC cells were analyzed to evaluate the effect of CG200745 or SAHA for 24 and 48 h.Cells were analyzed on Western blotting with cyclin D1, p21, caspase-3, c-myc, acetyl-H4, H3, and β-actin antibodies. Cell lysates of drug-treated A549, Calu6 (A), H358, and H596 (B) cells were compared to corresponding untreated cells.
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pone.0119379.g006: Changes in protein expression of NSCLC cells were analyzed to evaluate the effect of CG200745 or SAHA for 24 and 48 h.Cells were analyzed on Western blotting with cyclin D1, p21, caspase-3, c-myc, acetyl-H4, H3, and β-actin antibodies. Cell lysates of drug-treated A549, Calu6 (A), H358, and H596 (B) cells were compared to corresponding untreated cells.

Mentions: To further confirm the correlation of mRNA expression and changes in H4K16ac after CG200745 treatment, mRNA expression of the CCND1 and p21 genes were assessed. The H4K16ac levels of the CCND1 and p21 -promoter region were analyzed in untreated (control) and CG200745-treated (CG5) Calu6 cells using a whole gene probe microarray. As shown in Fig. 5, the H4K16ac level in CCND1 and p21 genes after CG200745 treatment, especially near the TSS position, was found to be significantly altered (Fig. 5A) and to be correlated with the expression of mRNA when quantitatively measured (Fig. 5B). Expression of Mxi1 was increased after 24 hours treatment of CG200745 on Semi quantitative analysis with two different sets of primers (Fig. 5C). The protein expression levels of cyclin D1 and p21 genes were dramatically reduced and increased, respectively, not only in Calu6 cells but also in several other NSCLC cells including A549, H358, and H596 cells (Fig. 6). In addition, CG200745 treatment resulted in activation of the intrinsic apoptotic caspase 3, in Calu6 cells as well as in several NSCLC cells including A549, H358, and H596 cells (Fig. 6A and 6B). Taken together, these data showed that the expression levels of certain genes were affected by CG200745 treatment through the H4K16ac level at the TSS region.


Epigenetic modulation with HDAC inhibitor CG200745 induces anti-proliferation in non-small cell lung cancer cells.

Chun SM, Lee JY, Choi J, Lee JH, Hwang JJ, Kim CS, Suh YA, Jang SJ - PLoS ONE (2015)

Changes in protein expression of NSCLC cells were analyzed to evaluate the effect of CG200745 or SAHA for 24 and 48 h.Cells were analyzed on Western blotting with cyclin D1, p21, caspase-3, c-myc, acetyl-H4, H3, and β-actin antibodies. Cell lysates of drug-treated A549, Calu6 (A), H358, and H596 (B) cells were compared to corresponding untreated cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4363698&req=5

pone.0119379.g006: Changes in protein expression of NSCLC cells were analyzed to evaluate the effect of CG200745 or SAHA for 24 and 48 h.Cells were analyzed on Western blotting with cyclin D1, p21, caspase-3, c-myc, acetyl-H4, H3, and β-actin antibodies. Cell lysates of drug-treated A549, Calu6 (A), H358, and H596 (B) cells were compared to corresponding untreated cells.
Mentions: To further confirm the correlation of mRNA expression and changes in H4K16ac after CG200745 treatment, mRNA expression of the CCND1 and p21 genes were assessed. The H4K16ac levels of the CCND1 and p21 -promoter region were analyzed in untreated (control) and CG200745-treated (CG5) Calu6 cells using a whole gene probe microarray. As shown in Fig. 5, the H4K16ac level in CCND1 and p21 genes after CG200745 treatment, especially near the TSS position, was found to be significantly altered (Fig. 5A) and to be correlated with the expression of mRNA when quantitatively measured (Fig. 5B). Expression of Mxi1 was increased after 24 hours treatment of CG200745 on Semi quantitative analysis with two different sets of primers (Fig. 5C). The protein expression levels of cyclin D1 and p21 genes were dramatically reduced and increased, respectively, not only in Calu6 cells but also in several other NSCLC cells including A549, H358, and H596 cells (Fig. 6). In addition, CG200745 treatment resulted in activation of the intrinsic apoptotic caspase 3, in Calu6 cells as well as in several NSCLC cells including A549, H358, and H596 cells (Fig. 6A and 6B). Taken together, these data showed that the expression levels of certain genes were affected by CG200745 treatment through the H4K16ac level at the TSS region.

Bottom Line: Histone modification plays a pivotal role on gene regulation, as regarded as global epigenetic markers, especially in tumor related genes.Treatment with CG200745 increased the global level of histone acetylation, resulting in the inhibition of cell proliferation.ChIP-on-chip analysis with an H4K16ac antibody showed altered H4K16 acetylation on genes critical for cell growth inhibition, although decreased at the transcription start site of a subset of genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Asan Medical Center, The University of Ulsan College of Medicine, Seoul, Republic of Korea.

ABSTRACT
Histone modification plays a pivotal role on gene regulation, as regarded as global epigenetic markers, especially in tumor related genes. Hence, chemical approaches targeting histone-modifying enzymes have emerged onto the main stage of anticancer drug discovery. Here, we investigated the therapeutic potentials and mechanistic roles of the recently developed histone deacetylase inhibitor, CG200745, in non-small cell lung cancer cells. Treatment with CG200745 increased the global level of histone acetylation, resulting in the inhibition of cell proliferation. ChIP-on-chip analysis with an H4K16ac antibody showed altered H4K16 acetylation on genes critical for cell growth inhibition, although decreased at the transcription start site of a subset of genes. Altered H4K16ac was associated with changes in mRNA expression of the corresponding genes, which were further validated in quantitative RT-PCR and western blotting assays. Our results demonstrated that CG200745 causes NSCLC cell growth inhibition through epigenetic modification of critical genes in cancer cell survival, providing pivotal clues as a promising chemotherapeutics against lung cancer.

No MeSH data available.


Related in: MedlinePlus