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Promoter hypomethylation up-regulates CD147 expression through increasing Sp1 binding and associates with poor prognosis in human hepatocellular carcinoma.

Kong LM, Liao CG, Chen L, Yang HS, Zhang SH, Zhang Z, Bian HJ, Xing JL, Chen ZN - J. Cell. Mol. Med. (2010)

Bottom Line: Significantly higher expression of CD147 and significantly lower promoter methylation level were observed in HCC cell lines compared to normal cell lines and tissues control.Moreover, HCC patients with unmethylated CD147 promoter had a significantly higher recurrence rate (88.1%versus 58.3%; P < 0.05) and death rate (83.3%versus 50.0%; P < 0.05) than patients with methylated CD147 promoter.In conclusions, promoter hypomethylation up-regulates CD147 expression primarily through increasing Sp1 binding and associates with poor prognosis in HCC patients.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Cancer Biology, Cell Engineering Research Centre & Department of Cell Biology, Fourth Military Medical University, Xi'an, China.

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Promoter hypomethylation up-regulated CD147 expression through increasing Sp1 binding in vivo. (A, B and C) The mRNA and protein expression of CD147 and Sp1 detected by regular RT-PCR, real-time quantitative RT-PCR and Western blot in QZG cell treated with different concentration of 5-Aza-dC. (D) Demethylation increased the binding of Sp1 to the CD147 promoter in vivo. ChIP assay using antibody against Sp1 was performed in before and after 5-Aza-dC treatment QZG cell. The normal rabbit IgG was used as a negative control and Input indicates 5% input DNA, a positive amplification control.
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fig04: Promoter hypomethylation up-regulated CD147 expression through increasing Sp1 binding in vivo. (A, B and C) The mRNA and protein expression of CD147 and Sp1 detected by regular RT-PCR, real-time quantitative RT-PCR and Western blot in QZG cell treated with different concentration of 5-Aza-dC. (D) Demethylation increased the binding of Sp1 to the CD147 promoter in vivo. ChIP assay using antibody against Sp1 was performed in before and after 5-Aza-dC treatment QZG cell. The normal rabbit IgG was used as a negative control and Input indicates 5% input DNA, a positive amplification control.

Mentions: To verify the role of DNA methylation in CD147 expression regulation, the QZG cell was treated with DNA-demethylating reagent 5-Aza-dC and then CD147 expression was examined. The mRNA and protein expression of CD147 were significantly increased in a dose-dependent manner after treatment (Fig. 4A–C). The methylation status in treated QZG cell was then determined by BGS. We found that the methylation level of CD147 promoter region was significantly decreased with the increased concentration of 5-Aza-dC in a dose-dependent manner (data not shown). Considering the possible role of Sp1 in the activation of CD147 transcription, to rule out the possibility of Sp1 involvement in 5-Aza-dC-induced CD147 up-regulation, we examined the Sp1 expression level in treated QZG cell. Our data showed that the 5-Aza-dC treatment did not change the mRNA and protein expression level of Sp1 (Fig. 4A–C).


Promoter hypomethylation up-regulates CD147 expression through increasing Sp1 binding and associates with poor prognosis in human hepatocellular carcinoma.

Kong LM, Liao CG, Chen L, Yang HS, Zhang SH, Zhang Z, Bian HJ, Xing JL, Chen ZN - J. Cell. Mol. Med. (2010)

Promoter hypomethylation up-regulated CD147 expression through increasing Sp1 binding in vivo. (A, B and C) The mRNA and protein expression of CD147 and Sp1 detected by regular RT-PCR, real-time quantitative RT-PCR and Western blot in QZG cell treated with different concentration of 5-Aza-dC. (D) Demethylation increased the binding of Sp1 to the CD147 promoter in vivo. ChIP assay using antibody against Sp1 was performed in before and after 5-Aza-dC treatment QZG cell. The normal rabbit IgG was used as a negative control and Input indicates 5% input DNA, a positive amplification control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4373337&req=5

fig04: Promoter hypomethylation up-regulated CD147 expression through increasing Sp1 binding in vivo. (A, B and C) The mRNA and protein expression of CD147 and Sp1 detected by regular RT-PCR, real-time quantitative RT-PCR and Western blot in QZG cell treated with different concentration of 5-Aza-dC. (D) Demethylation increased the binding of Sp1 to the CD147 promoter in vivo. ChIP assay using antibody against Sp1 was performed in before and after 5-Aza-dC treatment QZG cell. The normal rabbit IgG was used as a negative control and Input indicates 5% input DNA, a positive amplification control.
Mentions: To verify the role of DNA methylation in CD147 expression regulation, the QZG cell was treated with DNA-demethylating reagent 5-Aza-dC and then CD147 expression was examined. The mRNA and protein expression of CD147 were significantly increased in a dose-dependent manner after treatment (Fig. 4A–C). The methylation status in treated QZG cell was then determined by BGS. We found that the methylation level of CD147 promoter region was significantly decreased with the increased concentration of 5-Aza-dC in a dose-dependent manner (data not shown). Considering the possible role of Sp1 in the activation of CD147 transcription, to rule out the possibility of Sp1 involvement in 5-Aza-dC-induced CD147 up-regulation, we examined the Sp1 expression level in treated QZG cell. Our data showed that the 5-Aza-dC treatment did not change the mRNA and protein expression level of Sp1 (Fig. 4A–C).

Bottom Line: Significantly higher expression of CD147 and significantly lower promoter methylation level were observed in HCC cell lines compared to normal cell lines and tissues control.Moreover, HCC patients with unmethylated CD147 promoter had a significantly higher recurrence rate (88.1%versus 58.3%; P < 0.05) and death rate (83.3%versus 50.0%; P < 0.05) than patients with methylated CD147 promoter.In conclusions, promoter hypomethylation up-regulates CD147 expression primarily through increasing Sp1 binding and associates with poor prognosis in HCC patients.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Cancer Biology, Cell Engineering Research Centre & Department of Cell Biology, Fourth Military Medical University, Xi'an, China.

Show MeSH
Related in: MedlinePlus