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目的验证抑癌基因PTEN在肝细胞性肝癌(HCC)中的热点突变,探讨PTEN外显子基因突变与HCC的关系。方法收集经病理证实的HCC患者外周血和血清标本各100份。提取全血DNA,对PTEN突变热点所在的外显子1、4、5、6、7、8进行PCR扩增测序,SeqMan软件分析测序。通过Western blotting检测突变标本血清中PTEN蛋白的表达情况,并通过ELISA检测HCC血清PTEN蛋白表达情况。结果 100份PTEN外显子标本中共发现14个碱基改变,其中7个位于外显子区域,7个位于内含子区域。Western blotting结果显示突变标本的PTEN蛋白表达明显下降,与血清ELISA的结果一致,说明突变是引起HCC患者外周血中PTEN蛋白表达下降的原因之一。结论 PTEN基因8号外显子是HCC的突变热点,但PTEN在HCC中突变频率不高。PTEN基因突变是PTEN蛋白表达异常的原因之一,但不是主要影响因素。
Objective To verify the hotspot mutation of tumor suppressor gene PTEN in hepatocellular carcinoma (HCC) and explore the relationship between PTEN exon mutation and HCC. Methods 100 peripheral blood and serum samples were collected from pathologically confirmed HCC patients. Whole blood DNA was extracted, and the exons 1, 4, 5, 6, 7, 8 of the PTEN mutation hotspot were subjected to PCR amplification and sequencing, and SeqMan software was used for analysis and sequencing. The expression of PTEN protein in the serum of the mutant samples was detected by Western blotting, and the expression of PTEN protein in HCC serum was detected by ELISA. Results A total of 14 bases were found in 100 PTEN exons, of which 7 were located in the exon region and 7 in the intron region. Western blotting showed that the expression of PTEN protein in the mutant samples was significantly decreased, which was consistent with the results of serum ELISA. It indicated that mutation was one of the causes of the decrease of PTEN protein expression in peripheral blood of patients with HCC. Conclusion The exon 8 of PTEN gene is a hotspot of HCC mutation, but the frequency of PTEN mutation in HCC is not high. PTEN gene mutation is one of the causes of abnormal expression of PTEN protein, but it is not the main influence factor.