| Objective: Hepatocellular carcinoma(HCC)is a highly heterogeneous disease caused by different pathogenic factors and genetic variation.At present,some progress has been made in the study of HCC gene mutation,but the description of HCC gene mutation map is far from complete,and the key drivers of HCC are still lack of understanding.In this study,with the help of the next generation sequencing(NGS)technique and Illumina high throughput sequencing platform,the characteristics of oncogene mutation in patients with hepatocellular carcinoma(HCC)were analyzed,and the correlation between gene mutation and clinicopathological factors in patients with HCC was investigated.Methods: From January 2017 to December 2018,a total of 164 HCC patients underwent hepatectomy or puncture biopsy and gene detection of tumor tissues and peripheral blood samples in the affiliated Hospital of Qingdao University.Tissue samples of formalin-fixed paraffin-embedded(FFPE)for 4um-10 um 8-10 samples,and peripheral blood 200-500 ul as controls.Extraction of genomic DNA from tissue and peripheral blood samples by QIAGEN QIAamp○R DNA FFPE Tissue kit and QIAamp○R DNA Mini kit respectively.Genome analysis was carried out in the laboratory of OrigiMed(Shanghai,China),a CAP accredited lab.All the encoding exons of 450 key cancer-related genes were captured by the custom hybridization capture panel.In addition,the probe density was increased to ensure high efficiency of capture in the conservatively low read depths region.At the same time,the clinical and pathological data of the patients were collected.The relationship between gene mutation and clinicopathological factors was analyzed by univariate analysis and Logistic regression analysis.Results: In this study,there were 146 males and 18 females,with an average age of 56.3 years(27-78years).A total of 1097 mutated genes were detected in the tumor tissues of the patients,and the order of mutation frequency from high to low is TP53(57.9%),TERT(46.3%),CTNNB1(22.0%),AXIN1(13.3%),CCND1(13.1%),RB1(12.7%),TSC2(11.4%),et al.Mutation forms include single nucleotide variation,copy number variation,short or long segment insertion deletion,and gene rearrangement / fusion.The single nucleotide variation was the main form of variation,and the transformation of C:G>T:A is the main form of variation.P53 pathway,telomere repair pathway,WNT pathway,PI3 K / mTOR pathway and cell cycle related pathway were the most easily mutated carcinogenic pathways in the development of HCC.TP53 gene mutation was significantly correlated with Edmondson’s grade III-IV and microvascular invasion(P=0.00000 and P=0.000 respectively).TERT gene mutation was associated with Glisson capsule invasion(P=0.004).The simultaneous mutation of TP53 and TERT were associated with Edmondson’s grade III-IV,microvessel invasion,and Glisson capsule invasion(P=0.011,P=0.015 and P=0.006 respectively).The percentage of Edmondson’s grade I-II and AFP ≤ 20ng/mL in patients with CTNNB1 gene mutation was significantly higher than that in patients with no mutation in CTNNB1 gene(P < 0.023 and P < 0.002 respectively).Logistic regression analysis showed that Edmondson’s grade III-IV and microvascular invasion were risk factors for TP53 gene mutation(OR=2.807,95%CI: 1.412-5.580,P=0.003;OR=2.774,95% CI: 1.391-5.534,P=0.004).AFP ≤ 20ng/mL was a risk factor for CTNNB1 gene mutation(OR=0.402,95% CI: 0.182-0.888,P=0.024).Edmondson’s grade III-IV was a risk factor(OR=2.265,95%CI:1.084-4.729,P=0.030)for simultaneous mutation of TP53 and TERT genes.Conclusion:In this study,TP53,TERT and CTNNB1 were the most easily mutated genes in patients with HCC.P53 pathway,telomere repair pathway and Wnt pathway were the most easily mutated carcinogenic pathways in the development of HCC.Some clinicopathological features of HCC were closely related to the types of mutated genes.These findings will help us to transform the biological knowledge of HCC into clinical practice and provide a new target for individualized therapy of HCC. |
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