Aberrant TGFβ/SMAD4 signaling contributes to epigenetic silencing of a putative tumor suppressor, Runx1T1 in ovarian cancer

Kun Tu Yeh; Tze Ho Chen; Hui Wen Yang; Jian Liang Chou; Lin Yu Chen; Chia Ming Yeh; Yu Hsin Chen; Ru Inn Lin; Her Young Su; Gary C.W. Chen; Daniel E. Deatherage; Yi Wen Huang; Pearlly S. Yan; Huey Jen Lin; Kenneth P. Nephew; Tim H.M. Huang; Hung Cheng Lai; Michael W.Y. Chan

doi:10.4161/epi.6.6.15856

Aberrant TGFβ/SMAD4 signaling contributes to epigenetic silencing of a putative tumor suppressor, Runx1T1 in ovarian cancer

Kun Tu Yeh, Tze Ho Chen, Hui Wen Yang, Jian Liang Chou, Lin Yu Chen, Chia Ming Yeh, Yu Hsin Chen, Ru Inn Lin, Her Young Su, Gary C.W. Chen, Daniel E. Deatherage, Yi Wen Huang, Pearlly S. Yan, Huey Jen Lin, Kenneth P. Nephew, Tim H.M. Huang, Hung Cheng Lai, Michael W.Y. Chan

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Abstract

Aberrant TGFβ signaling pathway may alter the expression of down-stream targets and promotes ovarian carcinogenesis. However, the mechanism of this impairment is not fully understood. Our previous study has identified RunX1T1 as a putative SMAD4 target in an immortalized ovarian surface epithelial cell line, IOSE. In this study, we report that transcription of RunX1T1 was confirmed to be positively regulated by SMAD4 in IOSE cells and epigenetically silenced in a panel of ovarian cancer cell lines by promoter hypermethylation and histone methylation at H3 lysine 9. SMAD4 depletion increased repressive histone modifications of RunX1T1 promoter without affecting promoter methylation in IOSE cells. Epigenetic treatment can restore RunX1T1 expression by reversing its epigenetic status in MCP3 ovarian cancer cells. When transiently treated with a demethylating agent, the expression of RunX1T1 was partially restored in MCP3 cells, but gradual re-silencing through promoter re-methylation was observed after the treatment. Interestingly, SMAD4 knockdown accelerated this re-silencing process, suggesting that normal TGFβ signaling is essential for the maintenance of RunX1T1 expression. In vivo analysis confirmed that hypermethylation of RunX1T1 was detected in 35.7% (34/95) of ovarian tumors with high clinical stages (p = 0.035) and in 83% (5/6) of primary ovarian cancer-initiating cells. Additionally, concurrent methylation of RunX1T1 and another SMAD4 target, FBXO32 which was previously found to be hypermethylated in ovarian cancer was observed in this same sample cohort (p < 0.05). Restoration of RunX1T1 inhibited cancer cell growth. Taken together, dysregulated TGFβ/SMAD4 signaling may lead to epigenetic silencing of a putative tumor suppressor, RunX1T1, during ovarian carcinogenesis.

Original language	English (US)
Pages (from-to)	727-739
Number of pages	13
Journal	Epigenetics
Volume	6
Issue number	6
DOIs	https://doi.org/10.4161/epi.6.6.15856
State	Published - Jun 2011
Externally published	Yes

Keywords

Epigenetics
Ovarian cancer
RunX1T1
TGFβ

ASJC Scopus subject areas

Molecular Biology
Cancer Research

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10.4161/epi.6.6.15856

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Yeh, K. T., Chen, T. H., Yang, H. W., Chou, J. L., Chen, L. Y., Yeh, C. M., Chen, Y. H., Lin, R. I., Su, H. Y., Chen, G. C. W., Deatherage, D. E., Huang, Y. W., Yan, P. S., Lin, H. J., Nephew, K. P., Huang, T. H. M., Lai, H. C., & Chan, M. W. Y. (2011). Aberrant TGFβ/SMAD4 signaling contributes to epigenetic silencing of a putative tumor suppressor, Runx1T1 in ovarian cancer. Epigenetics, 6(6), 727-739. https://doi.org/10.4161/epi.6.6.15856

Yeh, KT, Chen, TH, Yang, HW, Chou, JL, Chen, LY, Yeh, CM, Chen, YH, Lin, RI, Su, HY, Chen, GCW, Deatherage, DE, Huang, YW, Yan, PS, Lin, HJ, Nephew, KP, Huang, THM, Lai, HC & Chan, MWY 2011, 'Aberrant TGFβ/SMAD4 signaling contributes to epigenetic silencing of a putative tumor suppressor, Runx1T1 in ovarian cancer', Epigenetics, vol. 6, no. 6, pp. 727-739. https://doi.org/10.4161/epi.6.6.15856

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title = "Aberrant TGFβ/SMAD4 signaling contributes to epigenetic silencing of a putative tumor suppressor, Runx1T1 in ovarian cancer",

abstract = "Aberrant TGFβ signaling pathway may alter the expression of down-stream targets and promotes ovarian carcinogenesis. However, the mechanism of this impairment is not fully understood. Our previous study has identified RunX1T1 as a putative SMAD4 target in an immortalized ovarian surface epithelial cell line, IOSE. In this study, we report that transcription of RunX1T1 was confirmed to be positively regulated by SMAD4 in IOSE cells and epigenetically silenced in a panel of ovarian cancer cell lines by promoter hypermethylation and histone methylation at H3 lysine 9. SMAD4 depletion increased repressive histone modifications of RunX1T1 promoter without affecting promoter methylation in IOSE cells. Epigenetic treatment can restore RunX1T1 expression by reversing its epigenetic status in MCP3 ovarian cancer cells. When transiently treated with a demethylating agent, the expression of RunX1T1 was partially restored in MCP3 cells, but gradual re-silencing through promoter re-methylation was observed after the treatment. Interestingly, SMAD4 knockdown accelerated this re-silencing process, suggesting that normal TGFβ signaling is essential for the maintenance of RunX1T1 expression. In vivo analysis confirmed that hypermethylation of RunX1T1 was detected in 35.7% (34/95) of ovarian tumors with high clinical stages (p = 0.035) and in 83% (5/6) of primary ovarian cancer-initiating cells. Additionally, concurrent methylation of RunX1T1 and another SMAD4 target, FBXO32 which was previously found to be hypermethylated in ovarian cancer was observed in this same sample cohort (p < 0.05). Restoration of RunX1T1 inhibited cancer cell growth. Taken together, dysregulated TGFβ/SMAD4 signaling may lead to epigenetic silencing of a putative tumor suppressor, RunX1T1, during ovarian carcinogenesis.",

keywords = "Epigenetics, Ovarian cancer, RunX1T1, TGFβ",

author = "Yeh, {Kun Tu} and Chen, {Tze Ho} and Yang, {Hui Wen} and Chou, {Jian Liang} and Chen, {Lin Yu} and Yeh, {Chia Ming} and Chen, {Yu Hsin} and Lin, {Ru Inn} and Su, {Her Young} and Chen, {Gary C.W.} and Deatherage, {Daniel E.} and Huang, {Yi Wen} and Yan, {Pearlly S.} and Lin, {Huey Jen} and Nephew, {Kenneth P.} and Huang, {Tim H.M.} and Lai, {Hung Cheng} and Chan, {Michael W.Y.}",

year = "2011",

month = jun,

doi = "10.4161/epi.6.6.15856",

language = "English (US)",

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pages = "727--739",

journal = "Epigenetics",

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T1 - Aberrant TGFβ/SMAD4 signaling contributes to epigenetic silencing of a putative tumor suppressor, Runx1T1 in ovarian cancer

AU - Yeh, Kun Tu

AU - Chen, Tze Ho

AU - Yang, Hui Wen

AU - Chou, Jian Liang

AU - Chen, Lin Yu

AU - Yeh, Chia Ming

AU - Chen, Yu Hsin

AU - Lin, Ru Inn

AU - Su, Her Young

AU - Chen, Gary C.W.

AU - Deatherage, Daniel E.

AU - Huang, Yi Wen

AU - Yan, Pearlly S.

AU - Lin, Huey Jen

AU - Nephew, Kenneth P.

AU - Huang, Tim H.M.

AU - Lai, Hung Cheng

AU - Chan, Michael W.Y.

PY - 2011/6

Y1 - 2011/6

N2 - Aberrant TGFβ signaling pathway may alter the expression of down-stream targets and promotes ovarian carcinogenesis. However, the mechanism of this impairment is not fully understood. Our previous study has identified RunX1T1 as a putative SMAD4 target in an immortalized ovarian surface epithelial cell line, IOSE. In this study, we report that transcription of RunX1T1 was confirmed to be positively regulated by SMAD4 in IOSE cells and epigenetically silenced in a panel of ovarian cancer cell lines by promoter hypermethylation and histone methylation at H3 lysine 9. SMAD4 depletion increased repressive histone modifications of RunX1T1 promoter without affecting promoter methylation in IOSE cells. Epigenetic treatment can restore RunX1T1 expression by reversing its epigenetic status in MCP3 ovarian cancer cells. When transiently treated with a demethylating agent, the expression of RunX1T1 was partially restored in MCP3 cells, but gradual re-silencing through promoter re-methylation was observed after the treatment. Interestingly, SMAD4 knockdown accelerated this re-silencing process, suggesting that normal TGFβ signaling is essential for the maintenance of RunX1T1 expression. In vivo analysis confirmed that hypermethylation of RunX1T1 was detected in 35.7% (34/95) of ovarian tumors with high clinical stages (p = 0.035) and in 83% (5/6) of primary ovarian cancer-initiating cells. Additionally, concurrent methylation of RunX1T1 and another SMAD4 target, FBXO32 which was previously found to be hypermethylated in ovarian cancer was observed in this same sample cohort (p < 0.05). Restoration of RunX1T1 inhibited cancer cell growth. Taken together, dysregulated TGFβ/SMAD4 signaling may lead to epigenetic silencing of a putative tumor suppressor, RunX1T1, during ovarian carcinogenesis.

AB - Aberrant TGFβ signaling pathway may alter the expression of down-stream targets and promotes ovarian carcinogenesis. However, the mechanism of this impairment is not fully understood. Our previous study has identified RunX1T1 as a putative SMAD4 target in an immortalized ovarian surface epithelial cell line, IOSE. In this study, we report that transcription of RunX1T1 was confirmed to be positively regulated by SMAD4 in IOSE cells and epigenetically silenced in a panel of ovarian cancer cell lines by promoter hypermethylation and histone methylation at H3 lysine 9. SMAD4 depletion increased repressive histone modifications of RunX1T1 promoter without affecting promoter methylation in IOSE cells. Epigenetic treatment can restore RunX1T1 expression by reversing its epigenetic status in MCP3 ovarian cancer cells. When transiently treated with a demethylating agent, the expression of RunX1T1 was partially restored in MCP3 cells, but gradual re-silencing through promoter re-methylation was observed after the treatment. Interestingly, SMAD4 knockdown accelerated this re-silencing process, suggesting that normal TGFβ signaling is essential for the maintenance of RunX1T1 expression. In vivo analysis confirmed that hypermethylation of RunX1T1 was detected in 35.7% (34/95) of ovarian tumors with high clinical stages (p = 0.035) and in 83% (5/6) of primary ovarian cancer-initiating cells. Additionally, concurrent methylation of RunX1T1 and another SMAD4 target, FBXO32 which was previously found to be hypermethylated in ovarian cancer was observed in this same sample cohort (p < 0.05). Restoration of RunX1T1 inhibited cancer cell growth. Taken together, dysregulated TGFβ/SMAD4 signaling may lead to epigenetic silencing of a putative tumor suppressor, RunX1T1, during ovarian carcinogenesis.

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Aberrant TGFβ/SMAD4 signaling contributes to epigenetic silencing of a putative tumor suppressor, Runx1T1 in ovarian cancer

Abstract

Keywords

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