Methylation detection of phosphatase and tensin homolog deleted on chromosome ten gene promoter in hepatocellular carcinoma samples by next-generation sequencing
10.3760/cma.j.cn112150-20210302-00208
- VernacularTitle:应用二代测序技术检测肝细胞癌组织样本中抑癌基因 PTEN基因启动子甲基化率
- Author:
Xiaokuan JING
1
;
Qiyu JIANG
;
Congshu LI
;
Nianrong ZHANG
;
Yantao CHAI
;
Fan FENG
;
Boan LI
;
Yankun LI
Author Information
1. 湖北科技学院药学院，咸宁 437100
- Keywords:
Methylation;
Hepatocellular carcinoma;
Next generation sequencing
- From:
Chinese Journal of Preventive Medicine
2021;55(10):1220-1227
- CountryChina
- Language:Chinese
-
Abstract:
Objective:The purpose of this study is to use the next-generation sequencing (NGS) technology platform to detect the methylation rate of phosphatase and tensin homolog deleted on chromosome ten ( PTEN) promoter region in hepatocellular carcinoma (HCC) tissue samples, and to analyze the clinical significance of its correlation with the prognosis of patients receiving sorafenib treatment. Methods:The 52 pairs of tumor tissue and para-cancerous tissue samples from HCC patients treated with sorafenib alone, which were collected and preserved in the Liver Tumor Diagnosis and Research Center of the former 302 Hospital of the People′s Liberation Army by the National Natural Science Foundation of China Youth Project with the project batch number 81702986 in 2018, were extracted total DNA from the samples. Then the DNA samples were treated with bisulfite and specific primers were designed to amplify the PTEN promoter region. Finally, the amplified products were analyzed by second-generation sequencing. In the analysis of clinical significance of PTEN methylation, log-rank statistical analysis was used to calculate whether there was a statistical difference in survival between the patient groups. Results:The methylation rate of PTEN promoter region in tumor tissues (29.17%±9.58%) was significantly higher than that in paracancer tissues (4.17%±2.86%)( t=19.970, P<0.05). At the same time, in HCC tissues, the methylation rate of the PTEN promoter region is negatively correlated with its expression ( F=47.270, P<0.000 1; Y=-1 800× X+38.03), and the PTEN methylation rate is negatively correlated with the prognosis of patients receiving the molecularly targeted drug Sorafenib (χ2=4.313, P<0.05). Conclusion:This study successfully established a new method for detecting methylation in the promoter region of PTEN, and the methylation rate of PTEN can be used as one of the targets of HCC diagnosis and targeted therapy.
