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Objective@#To investigate the molecular markers of copy number aberrations (CNAs) of genes related to extrohepatic metastasis-free survival after the operation for hepatocellular carcinoma (HCC).@*Methods@#The CNA status of 20 candidate genes in 66 HCC samples was detected by microarray comparative genomic hybridization. The associations between gene CNAs and extrohepatic metastasis-free survival were evaluated using the Cox regression model, Log-rank test, and Kaplan-Meier survival analysis.@*Results@#Multivariate Cox analysis revealed that the independent risk factors for metastasis-free survival were MDM4 gain (hazard ratio [HR] = 2.74, 95% confidence interval [CI] = 1.18-6.37, P < 0.05), APC loss (HR = 8.43, 95% CI = 2.48-28.66, P < 0.01), and BCL2L1 gain (HR = 3.45, 95% CI = 1.13-10.52, P < 0.05) and the independent protective factor was FBXW7 loss (HR = 0.32, 95% CI = 0.12-0.89, P < 0.05). By stepwise Cox regression analysis, three CNAs related to metastasis-free survival were screened out: MDM4 gain (HR = 2.71, 95% CI = 1.11-6.64, P < 0.05), APC loss (HR = 7.19, 95% CI = 1.88-27.60, P < 0.005), and FBXW7 loss (HR = 0.16, 95% CI = 0.05-0.46, P < 0.01). There were significant differences in metastasis-free survival rate between the HCC patients with FBXW7 loss and without MDM4 gain or APC loss, those with MDM4 gain and/or APC loss and without FBXW7 loss, and those with other CNA combinations (log-rank test, P < 0.01).@*Conclusion@#MDM4 gain, APC loss, and FBXW7 loss are the independent prognostic factors for extrohepatic metastasis-free survival after the operation for HCC and can be used to predict the risk of extrohepatic metastasis after the operation for HCC.
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Objective To investigate the relationship between chromosome 6p copy number alterations (CNAs) and postoperative intrahepatic recurrence of hepatocellular carcinoma (HCC);and to screen for the target genes in CNA(s).Methods Array comparative genomic hybridization (CGH) and expression arrays were used to detect CNAs and differences in gene expression, respectively.The associations between CNAs in 6p and HCC recurrence were analyzed using the log-rank test, the Kaplan-Meier curves and the Cox proportional hazards models on 66 patients who had been follow-up for 2.6 ～ 73.3 months.The differentially expression of genes in the potentially recurrence-related CNAs were further evaluated by the MannWhitney U test on 117 HCCs, which included 109 cases with paired array CGH and expression data.Results 6p CNAs were detected in 46 (69.7％) of the 66 HCCs.Of the 8 CNAs with the most frequent recurrence of over 20％ , a gain at 6p21.1 was independently associated with a 2.3-fold (95％ CI =1.1 ～ 5.1, P ＜ 0.05) increased risk for intrahepatic recurrence and with a more pronounced 3.3-fold (95％ CI =1.4 ～ 8.2, P ＜0.05) risk for early recurrence (≤ 1 year).A panel of 9 genes, including BYSL and RPL7L1 within the documented 6p21.1, were observed to be upregulated in HCCs with 6p21.1 gain when compared with HCCs without (all P ＜ 0.05).A high BYSL expression significantly correlated with a larger tumor size (＞ 6 cm), vascular invasion and advanced tumor stage (all P ＜ 0.05), and high RPL7L1 expression significantly correlated with vascular invasion and advanced tumor stage (all P ＜ 0.05).Conclusion A gain at 6p21.1 was an independently prognostic marker for intrahepatic recurrence of postoperative HCC, particular for early recurrence, and BYSL and RPL7L1 might be the target genes in the recurrence-related 6p21.1 gain.
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OBJECTIVE To assess the association of copy number variations (CNVs) in chromosome 17q with the overall survival(OS) of patients with hepatocellular carcinoma(HCC), and to screen for target genes contained in the OS-related CNVs. METHODS A total of 174 HCC cases were enrolled. For 66 patients, the follow-up data was available. High-resolution Agilent Hu-244A array comparative genomic hybridization (aCGH) and Affymetrix U133 Plus 2.0 expression arrays were used to detect CNVs and gene expression of genes from the 17q region, respectively. The association of CNVs and OS was assessed with Log-rank test, Kaplan-Meier survival analysis, and Cox proportional hazards models. The gene expression in HCCs with 17q gain, HCCs without, and non-tumor liver tissues were compared with a Mann-Whitney U test. RESULTS Univariate association analysis showed that copy number gain in 17q25.1-25.3 was significantly associated with reduced OS (Log-rank test, P = 0.00002), and HCC cases with 17q25.1-25.3 gain had a 4.76-fold (95%CI: 2.31-9.81) increased hazard ratio (HR) for death from HCC, as compared to those without the gain. Multivariate Cox proportional hazards regression model revealed 17q25.1-25.3 gain to be an independent prognostic marker for poor OS (HR = 3.17, 95%CI: 1.39-7.26, P = 0.006). The expression levels of 18 genes in 17q25.1-25.3 including SLC9A3R1, GRB2, and TK1 were significantly increased in HCCs with gain than in those without (all P < 0.01) and non-tumor liver tissues (all P < 0.01). CONCLUSION The association of 17q25.1-25.3 gain with reduced OS has indicated that it is a prognostic marker for poor patient survival in HCC, for which SLC9A3R1, GRB2, and TK1 are candidate genes.
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Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Genetics , Mortality , Chromosomes, Human, Pair 17 , DNA Copy Number Variations , Liver Neoplasms , Genetics , MortalityABSTRACT
BACKGROUND: What role tumor-related genes play in the process of tumor generation, development, metastasis and prognosis has always been a thorny issue in medical field?OBJECTIVE: To study the detection of gene mutation in tumor by denaturing gradient gel electrophoresis(DGGE) and automated DNA se+uence analysis and the change of p53 gene and p53 protein during the development and metastasis of colorectal carcinoma so as to provide basis for evaluating the prognosis of colorectal carcinoma.DESIGN: Single sample study using the tissue specimen as subject.SETTING: Department of oncology in an affiliated hospital of a military medical university.PARTICIPANTS: We collected the primary focus and liver metastasis focus specimens from 41 patients with colon cancer who had hepatectomy because of liver metastasis 5 months to 5 years after radical operation for coloncancer. They were inpatients in Nanfang Hospital, First Military Medical University of Chinese PLA, from January 1994 to December 2000.METHODS: p53 gene(exons 5- 11) mutation of primary focus and liver metastasis focus specimens from 41 cases of colon cancer was examined by DGGE and automated DNA sequencing. Expression of p53 protein was detected by immunohistochemistry.histochemical staining.MAIN OUTCOME MEASURES:①Detection of the mutation of p53 gene by DGGE;②Analysis of p53 gene sequence;③Results of p53 immunohistochemical staining.RESULTS: p53 gene mutation was detected in exons 5 - 9 in 24 out of 41patients(62% ) . Among them, 6 patients had p53 mutation in liver metastasis. The others had consistent mutations in both primary coloreetal and hepatic metastatic lesions. In addition, p53 mutation was also found in the metastatic lesion in three patients. Among the 16 cases of mutation in primary colorectal and hepatic metastatic lesions, 14 cases showed that the ratio of p53 base peak to normal peak was significantly higher in hepatic metastatic lesions than in primary colorectal lesions(P ＜ 0. 001) . Results of p53 immunohistochemical staining were highly consistent with those of DGGE and DNA sequence analysis. However, gene analysis detected focus with nonsense mutation while immunohistochemistry detected overexpression of p53 protein.CONCLUSION: p53 mutation, in patients with colorectal carcinoma followed by hepatic metastases, mostly originates from primary colorectal lesion and then is kept and metastasizes into hepatic cells. The amount of mutated p53 gene and the number of tumor cells containing p53 mutation are increased in hepatic metastatic lesion. P53 mutation is positively correlated with overexpression of p53 protein.
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To investigate the expression of cytokeratin 18(CYK18) in hepatocellular carcinoma(HCC) cell membrane and the relationship between the serum levels of tissue polypeptide specific antigen (TPS) and the proliferating activity of HCC cells. The serum levels of TPS were determined by TPS TM ELISA in 96 patients with HCC. Four micrometer paraffin embeded sections of liver specimens from these patients were immunostained with the strept avidin biotin complex immunoperoxidase technique (IHC SABC). All of the HCC preparations were positively stained by CYK 18 and proliferating cell nuclear antigen (PCNA) with IHC SABC.The results of PCNA stainings were: + in 62 5%, in 26 0%, in 8 3%,and in 3 1% .The median levels of TPS in the four groups were: + 137 98 U/L, 685 3 U/L, in 1 126 U/L and 4 672U/L respectively. There were significant differences among these groups ( P
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To investigate the value of tissue polypeptide specific antigen(TPS) in the diagnosis of hepatocellular carcinoma (HCC), the serum levels of TPS and alfa fetoprotein (AFP) were measured by TPS TM ELISA and enzyme immunoassay respectively in 96 patients with HCC, 30 patients with cirrhosis, 20 patients with hepatitis and 20 healthy controls.The diagnostic sensitivity of TPS and AFP is 89 6% and 73 0% respectively.There is a significant difference between them ( P
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To investigate the prognostic value of serum levels of tissue polypeptide specific antigen (TPS) in patients with hepatocellular carcinoma(HCC). Ninety six patients who had undergone hepatoma resections and 22 patients who received radio frequency ablations (RF) were followed up for 6～12 months.The serum levels of TPS were repeatedly determined.The RF group patients were divided into three subgroups: improved ,stable,and advanced.The median levels (U/L) of serum TPS in advanced subgroup were always higher than those in improved subgroup ( P