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 ...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 non-cancerous tissues of 20 HCC,3 normal liver tissues from 3 accidentally died healthy adults using simi-quantitatively Northerm blot.The methylation status was also assessed with methylation specfic PCR.RESULTS:p16 mRNA expression level was decressed 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<para-cancerous<non-cancerous liver tissues.p15 mRNA expression level was decreased in the cancerous tissues in 50%(10/20) HCC patients,of which one case had no p15 mRNA detected,4 cases (20%) displayed variation in the order of cancerous<para-cancerous<non-cancerous liver tissues.In cancerous,para-cancerous and non-cancerous tissues,p16 promoter CpG islands hypernethylation occurred 65% ,60% and 35%,while p15 promoter CpG islands hypermethylation occurred 50%,40% and 25%.of 12 HCCs with lower p16 promoter CpG islands methylation(91.6%).Hundred percent(10/10) HCCs with lower p15 mRNA expression level showed p15 promoter CpG islands methylation.Significant correlation between 5′CpG islands methylation and p16/p15 mRNA expression suppression was found.The dexreased expression of p16/p15 mRNA or metyylation of p16.p15 promoters 5′CpG island was significantly,0.01.2,0.00271,0.0218,respectively,(p<0.05).CONCLUSION:p16 and p15 genes transcriptional inactivation might play an important role in hepataocarcinogenesis.5′CpG islands methylation might bi the major mechanism of p16 and p15 genes inactivation in primary HCC in the studied population.5′CpG islands methylation of p16 and p15 genes might be an early event in hepatocarcinogenesis.展开更多
AIM: To study methylation, frequencies of homozygous deletion and mutation of p16 gene in gastric carcinoma.METHODS: The methylation pattern in exon 1 and exon 2of p16 gene was studied with polymerase chain reaction (...AIM: To study methylation, frequencies of homozygous deletion and mutation of p16 gene in gastric carcinoma.METHODS: The methylation pattern in exon 1 and exon 2of p16 gene was studied with polymerase chain reaction (PCR), using methylation sensitive restriction endonuclease HpaⅡ and methylation insensitive restriction endonudease MspⅠ. PCR technique was used to detect homozygous deletions of exon 1 and exon 2 of p16 gene and single strand conformation polymorphism (SSCP) technique was used to detect the mutation of the gene.RESULTS: Hypermethylation changes in exon 1 and exon 2 of p16 gene were observed in 25 % and 45 % of 20gastric cancer tissues, respectively, while no methylation abnormality was found in normal tissues. The homozygous deletion frequency of exon 1 and exon 2 of p16 gene in 20gastric cancer tissues was 20 % and 10 %, respectively. No mutation was found in exon 1 of p16 gene, while abnormal single strands were found in 2 (10 %) cases in exon 2 as detected by SSCP.CONCLUSION: The results suggest that hypermethylation and abnormality of p16 gene may play a key role in the progress of gastric cancer. Hypermethylation of exon 2 of p16 gene may have effects on the carcinogenesis of gastric mucosa and may be a later event.展开更多
基金Supported by the National Natural Science Foundation of China.No.39670702
文摘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 non-cancerous tissues of 20 HCC,3 normal liver tissues from 3 accidentally died healthy adults using simi-quantitatively Northerm blot.The methylation status was also assessed with methylation specfic PCR.RESULTS:p16 mRNA expression level was decressed 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<para-cancerous<non-cancerous liver tissues.p15 mRNA expression level was decreased in the cancerous tissues in 50%(10/20) HCC patients,of which one case had no p15 mRNA detected,4 cases (20%) displayed variation in the order of cancerous<para-cancerous<non-cancerous liver tissues.In cancerous,para-cancerous and non-cancerous tissues,p16 promoter CpG islands hypernethylation occurred 65% ,60% and 35%,while p15 promoter CpG islands hypermethylation occurred 50%,40% and 25%.of 12 HCCs with lower p16 promoter CpG islands methylation(91.6%).Hundred percent(10/10) HCCs with lower p15 mRNA expression level showed p15 promoter CpG islands methylation.Significant correlation between 5′CpG islands methylation and p16/p15 mRNA expression suppression was found.The dexreased expression of p16/p15 mRNA or metyylation of p16.p15 promoters 5′CpG island was significantly,0.01.2,0.00271,0.0218,respectively,(p<0.05).CONCLUSION:p16 and p15 genes transcriptional inactivation might play an important role in hepataocarcinogenesis.5′CpG islands methylation might bi the major mechanism of p16 and p15 genes inactivation in primary HCC in the studied population.5′CpG islands methylation of p16 and p15 genes might be an early event in hepatocarcinogenesis.
基金the National Natural Science Foundation of China,No.39170440
文摘AIM: To study methylation, frequencies of homozygous deletion and mutation of p16 gene in gastric carcinoma.METHODS: The methylation pattern in exon 1 and exon 2of p16 gene was studied with polymerase chain reaction (PCR), using methylation sensitive restriction endonuclease HpaⅡ and methylation insensitive restriction endonudease MspⅠ. PCR technique was used to detect homozygous deletions of exon 1 and exon 2 of p16 gene and single strand conformation polymorphism (SSCP) technique was used to detect the mutation of the gene.RESULTS: Hypermethylation changes in exon 1 and exon 2 of p16 gene were observed in 25 % and 45 % of 20gastric cancer tissues, respectively, while no methylation abnormality was found in normal tissues. The homozygous deletion frequency of exon 1 and exon 2 of p16 gene in 20gastric cancer tissues was 20 % and 10 %, respectively. No mutation was found in exon 1 of p16 gene, while abnormal single strands were found in 2 (10 %) cases in exon 2 as detected by SSCP.CONCLUSION: The results suggest that hypermethylation and abnormality of p16 gene may play a key role in the progress of gastric cancer. Hypermethylation of exon 2 of p16 gene may have effects on the carcinogenesis of gastric mucosa and may be a later event.
