【摘要】 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。更多还原

【Abstract】 Objective To Validate the hot spot mutations of tumor suppressor gene PTEN in hepatocellular carcinoma(HCC),and study the relationship of PTEN gene mutation in exon and HCC.Methods One hundred peripheral blood and serum samples of HCC patients pathologically proved were collected respectively.DNA was extracted from blood samples,the PTEN mutation hot spots located in exon 1,4,5,6,7,8for PCR amplification and sequencing,and analysis of SeqMan software.Through the Western blotting,the expression of PTEN protein with mutation in serum was detected,and the expression of PTEN protein in serum was detected with ELISA.Results Fourteen bases changed in the 100 PTEN specimens,seven of them located in exon region,and seven located in the intron region.Western blotting results showed that protein of PTEN with mutation was significantly decreased,which was consistent with the results of serum ELISA.It illustrated that the mutation was one of the reasons for the decline of expression of PTEN protein in the peripheral blood of HCC patients.ConclusionPTEN gene exon 8is the hot spot mutation in HCC,but PTEN in hepatocellular carcinoma mutation frequency is not high.Mutation of PTEN gene is one of the causes of abnormal expression of PTEN protein,but not the main influencing factors.更多还原