Effect of 5-2Aza-2'-deoxycytidine on the P16 tumor suppressor gene in hepatocellular carcinoma cell line HepG2

Liu, Li Hua

doi:10.3748/wjg.v7.i1.131

Cited by 23 publications

(11 citation statements)

References 39 publications

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“…It has been reported that higher expression of methyltransferases was associated with methylation of p16 and p15 genes in HCC [29] . It has been noted that either the inhibitors of methyltransferase, such as 5-aza-2'-deoxycytidine and tea-polyphenol (-)-epigallocatechin-3-gallate or antisense inhibitors of DNA methyltransferases, reactivated silenced tumor suppressor genes and inhibited the growth of cancer cells [30][31][32][33] . Thus, methylation of 5'CpG islands of p16 and p15 genes might be an early event in hepatocarcinogenesis.…”

Section: Discussionmentioning

confidence: 99%

Association of low p16INK4a and p15INK4b mRNAs expression with their CpG islands methylation with human hepatocellular carcinogenesis

Qin¹,

Liu²,

Li³

et al. 2004

WJG

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AIM:To study the significance of p16 and p15 transcription suppression with hypermethylation of their genes' 5'CpG islands during human hepatocellular carcinogenesis. METHODS:The mRNA expression levels of p16 and p15 genes were evaluated in cancerous, para-cancerous and noncancerous tissues of 20 HCC, 3 normal liver tissues from 3 accidentally died healthy adults using semi-quantitatively Northern blot. The methylation status was also assessed with methylation specific PCR. RESULTS: p16 mRNA expression level was decreased in the cancerous tissues in 60% (12/20) of HCC patients, of which 2 cases had no p16 mRNA detected, 5 cases (25%) displayed variation in the order of cancerous

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Section: Discussionmentioning

confidence: 99%

Association of low p16INK4a and p15INK4b mRNAs expression with their CpG islands methylation with human hepatocellular carcinogenesis

Qin¹,

Liu²,

Li³

et al. 2004

WJG

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“…Frozen tissue samples were ground into very fine powder in liquid nitrogen, suspended in lysis buffer and treated with proteinase K. DNA was extracted by phenol-chloroformisoamyl alcohol and ethanol precipitation [21] . Adjacent normal liver tissues were used as corresponding normal controls.…”

Section: Dna Extractionmentioning

confidence: 99%

Hemizygous deletion and hypermethylation of RUNX3 gene in hepatocellular carcinoma

Liu²

2004

WJG

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AIM:To analyze the genetic and epigenetic alterations of RUNX3 gene, a potential putative tumor suppressor gene, in hepatocellular carcinoma (HCC). METHODS:PCR-based loss of heterozygosity (LOH) detection, analysis of mutation with PCR-single strand conformational polymorphism (SSCP) and sequencing, and methylation study with methylation specific PCR (MSP) were performed on RUNX3 gene in a series of 62 HCCs along with their matched normal tissues. RESULTS:Mutation of RUNX3 gene was not found, but one single nucleotide polymorphism with T to A transversion at the second nucleotide of the 18th condon was found. Nine of 26 informative cases (34.6%) showed allelic loss on the polymorphic site and 30 cases (48.4%) revealed hypermethylation of RUNX3 gene in promoter CpG islands. Furthermore, of the 9 cases with LOH, 8 (88.9%) also had hypermethylation. CONCLUSION:Our findings indicate that inactivation of RUNX3 gene through allelic loss and promoter hypermethylation might be one of the major mechanisms in hepatocellualr carcinogenesis.Xiao WH, Liu WW. Hemizygous deletion and hypermethylation of RUNX3 gene in hepatocellular carcinoma.

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“…5-Aza-2'-deoxycytidine (5-Aza-CdR) is thought to act by inhibiting DNA methyltransferase that methylates cytosine residues in eukaryotic DNA (8). Several researchers have used it as an experimental tool for demethylation (9,10). This drug appears to be incorporated into the cellular DNA where it acts as a non-competitive inhibitor of the maintenance methylase (9,11).…”

Section: Introductionmentioning

confidence: 99%

Effect of 5-Aza-2'-deoxycytidine on SLC22A18 in glioma U251 cells

Chu

Feng

et al. 2011

Mol Med Report

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Abstract. SLC22A18 [solute carrier family 22 (organic cation transporter) member 18] is located within the 11p15.5 cluster, and may be a new tumor suppressor gene; evidence of SLC22A18 hypermethylation is documented in several types of human cancers. In order to determine whether SLC22A18 hypermethylation is involved in glioma, we determined the SLC22A18 gene protein expression, mRNA expression and methylation status in glioma U251 cells before and after treatment with 5-Aza-2'-deoxycytidine (5-Aza-CdR), and observed the change in growth. Glioma U251 cells treated with 5-AzaCdR were analyzed by flow cytometry to identify any change in their cell cycle profiles. Tumors induced via the injection of untreated U251 cells were measured. Immunohistochemistry, reverse transcriptase-polymerase chain reaction (RT-PCR) and PCR-based methylation assay were carried out to determine SLC22A18 gene protein expression, mRNA expression and methylation status in glioma U251 cells before and after treatment with 5-Aza-CdR. The treated cells showed an increase in their proportion in G1, from 79.2 to 83.5%, and a decrease in S phase, from 12.4 to 5.8%. The apoptotic rate increased from 6.4 to 15.8%. Tumors induced via the injection of untreated U251 cells were approximately 1.46 cm 3 in size, whereas the tumors induced by U251 cells treated with 5-Aza-CdR averaged 0.88 cm 3 in size. The expression levels of SLC22A18 protein and mRNA in U251 cells were increased following treatment with 5x10 -7 M 5-Aza-CdR. Prior to 5-Aza-CdR treatment, the SLC22A18 gene demonstrated hypermethylation and therefore could not be cleaved by HpaII and MspI. It is known that only the DNA digested with HpaII or MspI can be amplified. Following treatment with 5-Aza-CdR, the SLC22A18 gene became demethylated, and could then be cleaved by both of the enzymes, and this failed to be amplified. 5-Aza-cdR may induce glioma U251 cell division and apoptosis and enhance demethylation and protein and mRNA expression of SLC22A18. The hypermethylation of SLC22A18 may be related to the transcriptional silencing of this gene. The growth inhibitory effects of 5-Aza-CdR treatment in vivo remain recognizable.

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Effect of 5-2Aza-2'-deoxycytidine on the P16 tumor suppressor gene in hepatocellular carcinoma cell line HepG2

Cited by 23 publications

References 39 publications

Association of low p16INK4a and p15INK4b mRNAs expression with their CpG islands methylation with human hepatocellular carcinogenesis

Association of low p16INK4a and p15INK4b mRNAs expression with their CpG islands methylation with human hepatocellular carcinogenesis

Hemizygous deletion and hypermethylation of RUNX3 gene in hepatocellular carcinoma

Effect of 5-Aza-2'-deoxycytidine on SLC22A18 in glioma U251 cells

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