Study On Genetic Susceptibility Of The Polymorphism And The Protein Experssion Of PTEN Gene With Familial Clustering Hepatocellular Carcinoma

Objective:(1)To investigate the potential correlation between the polymorphism and the protein expression of gene PTEN and genetic Susceptibilitin in Familial Clustering Hepatocellular Carcinoma; (2)To investigate the correlation between the polymorphism of PTEN gene and the prognosis of HCC(Hepatocelluar Carcinoma) patients.Materials and Methods :( 1 ) Immunohistochemical staining were employed to Detect the protein expression of PTEN genes in HCC tissues and the corresponding para-carcinoma tissues(para-HCC)from 53 cases of familial clustering HCC patients,60 cases of non-familial clustering HCC patients and in normal liver tissues from 65 cases of non-tumor patients;(2)Polymerase chain reaction -restriction fragment length polymorphism(PCR-RFLP)were employed to detect the three frequent genotype of single nucleotide polymorphisms(SNP) of the PTEN gene;(3)Check the detected genetype using DNA norientation sequencing ;(4)Analyze the prognosis of HCC(Hepatocelluar Carcinoma) patients after surgery.Results:(1)Immunohistochemical staining showed that the expression PTEN protein(2.28±1.03) in familial clustering HCC tissue was significantly lower than tha(t2.73±1.06)in non-familial clustering HCC and tha(t3.71±1.11) in normal liver tissue of non-tumor patients,and they are significantly different with each other(all were p<0.05);(2)the expression PTEN protein(2.52±1.06)in HCC(including familial clustering HCC and non-familial clustering HCC ,FH group+NFH group)tissue was also significantly lower than that(3.13±1.25)in para-HCC tissues and that(3.71±1.11)in normal liver tissue of non-tumor patients,and they are significantly different with each other(all were p<0.05), and that their positive expression level was obviously decreased along with pathology grade raising of HCC tissue; (3) The expression PTEN protein(2.96±1.24) in familial clustering HCC para-HCC tissues is not different with that(3.28±1.25)in non-familial clustering HCC para-HCC tissues in statistics,but they are significantly lower than that(3.71±1.11) in normal liver tissue of non-tumor patients;(4)There is no statistically significance SNP(rs2943772)polymorphism in PTEN gene(PTEN-A)in the three group. (5)C/C genotypic frequency of SNP(rs9651492)polymorphism in PTEN gene(PTEN-B)in the FH group(49.1%)was significantly higher than tha(t26.7%)in the NFH group and that(26.2%) in the N group,and C/C genotypic frequency was increase along with the increasing numbers of HCC patient in familial history, (P<0.05);The frequencies of PTEN-B(rs9651492) C/T genotype in HCC group(FH group 22.6%+NFH group 33.3%)was significantly lower than that of control group(N group52.3%).(6)The frequencies of PTEN-C(rs112025902) C/G genotype in HCC group (FH group 50.9%+NFH group 46.7%)was significantly higher than that of control group(N group27.7%), (P<0.05). (7)Correlation analysis showed that PTEN-B(rs9651492) genotype and the protein expression of PTEN gene took on obviously negative correlation for HCC family history,the numbers of HCC patients in familial history,pathology grade of HCC(correlation quotiety between-0.350～-0.158, P<0.05); (8)Contrasting the DNA sequencing consequence with PTEN gene order in GeneBank, verificated the PCR amplification part is the same with that of experiment design.(9)one annual survival rate of the patients taking along with PTEN-B(rs9651492)C/C genotype is lower than that taking along with PTEN-B(rs9651492)T/T genotype or C/T genotype in familial clustering hepatocellular carcinoma.Conclusion: (1)The protein expression of PTEN in HCC tissues and the corresponding para-carcinoma tissues(para-HCC) is low,and the protein expression of PTEN is lower in HCC with family HCC history. (2) the polymorphism of PTEN-B(rs9651492) site existed affinity with familial clustering HCC , thus showed that the PTEN-B(rs9651492) C/C genotype might be the genetic susceptive genotype of the familial clustering HCC.(3)the expression PTEN protein in HCC tissue was significantly lower than that in normal liver tissue of non-tumor patients, the expression PTEN protein in HCC tissue decrease along with increasing of pathology grade,this showed the expression PTEN protein might existed affinity with the biological behaviour of HCC; (4)In familial clustering hepatocellular carcinoma patients,the patient who take along PTEN-B(rs9651492) C/C genotype hinted that prognosis was bad.