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EVI1, a target gene for amplification at 3q26, antagonizes transforming growth factor-β-mediated growth inhibition in hepatocellular carcinoma.

Yasui K, Konishi C, Gen Y, Endo M, Dohi O, Tomie A, Kitaichi T, Yamada N, Iwai N, Nishikawa T, Yamaguchi K, Moriguchi M, Sumida Y, Mitsuyoshi H, Tanaka S, Arii S, Itoh Y - Cancer Sci. (2015)

Bottom Line: Knockdown of EVI1 resulted in increased induction of the cyclin-dependent kinase inhibitor p15(INK) (4B) by transforming growth factor (TGF)-β and decreased expression of c-Myc, cyclin D1, and phosphorylated Rb in TGF-β-treated cells.Consequently, knockdown of EVI1 led to reduced DNA synthesis and cell viability.Collectively, our results suggest that EVI1 is a probable target gene that acts as a driving force for the amplification at 3q26 in HCC and that the oncoprotein EVI1 antagonizes TGF-β-mediated growth inhibition of HCC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine, Kyoto, Japan.

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DNA copy number of EVI1, mRNA levels of EVI1, MDS1–EVI1, and MDS1, and EVI1 protein expression in hepatocellular carcinoma (HCC) cell lines. (a) DNA copy number of EVI1 in 20 HCC cell lines and four normal peripheral blood lymphocytes as measured by quantitative PCR. Values are normalized such that the average copy number in genomic DNA derived from four normal lymphocytes has a value of 1 (solid horizontal line). A value corresponding to the mean + two SD of the copy number of normal lymphocytes was used as the cut-off value for copy number gain (dotted line). Asterisks indicate cell lines showing copy number gain. (b) Relative levels of EVI1, MDS1–EVI1, and MDS1 mRNAs in 18 HCC cell lines as determined by quantitative TaqMan PCR. (c) Immunoblot analyses of EVI1 protein in the indicated cell lines. β-actin was blotted as an internal control.
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fig02: DNA copy number of EVI1, mRNA levels of EVI1, MDS1–EVI1, and MDS1, and EVI1 protein expression in hepatocellular carcinoma (HCC) cell lines. (a) DNA copy number of EVI1 in 20 HCC cell lines and four normal peripheral blood lymphocytes as measured by quantitative PCR. Values are normalized such that the average copy number in genomic DNA derived from four normal lymphocytes has a value of 1 (solid horizontal line). A value corresponding to the mean + two SD of the copy number of normal lymphocytes was used as the cut-off value for copy number gain (dotted line). Asterisks indicate cell lines showing copy number gain. (b) Relative levels of EVI1, MDS1–EVI1, and MDS1 mRNAs in 18 HCC cell lines as determined by quantitative TaqMan PCR. (c) Immunoblot analyses of EVI1 protein in the indicated cell lines. β-actin was blotted as an internal control.

Mentions: The DNA copy number of EVI1 was then determined in the 20 HCC cell lines by using quantitative PCR. The sequence-tagged site marker, SHGC-77524, which is specific for EVI1, was used for this PCR (Fig.1b). For this analysis, copy number changes were counted as gains if the copy number for a given tumor cell line exceeded the mean plus two standard deviations of that in normal lymphocytes. A copy number gain of EVI1 was observed in seven (35%) of the 20 cell lines (Fig.2a). As expected, JHH-1 cells showed the highest copy number gain.


EVI1, a target gene for amplification at 3q26, antagonizes transforming growth factor-β-mediated growth inhibition in hepatocellular carcinoma.

Yasui K, Konishi C, Gen Y, Endo M, Dohi O, Tomie A, Kitaichi T, Yamada N, Iwai N, Nishikawa T, Yamaguchi K, Moriguchi M, Sumida Y, Mitsuyoshi H, Tanaka S, Arii S, Itoh Y - Cancer Sci. (2015)

DNA copy number of EVI1, mRNA levels of EVI1, MDS1–EVI1, and MDS1, and EVI1 protein expression in hepatocellular carcinoma (HCC) cell lines. (a) DNA copy number of EVI1 in 20 HCC cell lines and four normal peripheral blood lymphocytes as measured by quantitative PCR. Values are normalized such that the average copy number in genomic DNA derived from four normal lymphocytes has a value of 1 (solid horizontal line). A value corresponding to the mean + two SD of the copy number of normal lymphocytes was used as the cut-off value for copy number gain (dotted line). Asterisks indicate cell lines showing copy number gain. (b) Relative levels of EVI1, MDS1–EVI1, and MDS1 mRNAs in 18 HCC cell lines as determined by quantitative TaqMan PCR. (c) Immunoblot analyses of EVI1 protein in the indicated cell lines. β-actin was blotted as an internal control.
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Related In: Results  -  Collection

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fig02: DNA copy number of EVI1, mRNA levels of EVI1, MDS1–EVI1, and MDS1, and EVI1 protein expression in hepatocellular carcinoma (HCC) cell lines. (a) DNA copy number of EVI1 in 20 HCC cell lines and four normal peripheral blood lymphocytes as measured by quantitative PCR. Values are normalized such that the average copy number in genomic DNA derived from four normal lymphocytes has a value of 1 (solid horizontal line). A value corresponding to the mean + two SD of the copy number of normal lymphocytes was used as the cut-off value for copy number gain (dotted line). Asterisks indicate cell lines showing copy number gain. (b) Relative levels of EVI1, MDS1–EVI1, and MDS1 mRNAs in 18 HCC cell lines as determined by quantitative TaqMan PCR. (c) Immunoblot analyses of EVI1 protein in the indicated cell lines. β-actin was blotted as an internal control.
Mentions: The DNA copy number of EVI1 was then determined in the 20 HCC cell lines by using quantitative PCR. The sequence-tagged site marker, SHGC-77524, which is specific for EVI1, was used for this PCR (Fig.1b). For this analysis, copy number changes were counted as gains if the copy number for a given tumor cell line exceeded the mean plus two standard deviations of that in normal lymphocytes. A copy number gain of EVI1 was observed in seven (35%) of the 20 cell lines (Fig.2a). As expected, JHH-1 cells showed the highest copy number gain.

Bottom Line: Knockdown of EVI1 resulted in increased induction of the cyclin-dependent kinase inhibitor p15(INK) (4B) by transforming growth factor (TGF)-β and decreased expression of c-Myc, cyclin D1, and phosphorylated Rb in TGF-β-treated cells.Consequently, knockdown of EVI1 led to reduced DNA synthesis and cell viability.Collectively, our results suggest that EVI1 is a probable target gene that acts as a driving force for the amplification at 3q26 in HCC and that the oncoprotein EVI1 antagonizes TGF-β-mediated growth inhibition of HCC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine, Kyoto, Japan.

Show MeSH
Related in: MedlinePlus