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AIM:To characterize and evaluate DNA alterations among intrahepatic cholangiocarcinoma (ICC) patients. METHODS:DNA from tumor and corresponding normal tissues of 52 patients was amplified with 33 arbitrary primers. The DNA fragment that alters most frequently in ICC was cloned,sequenced,and identified by comparison with known nucleotide sequences in the genome database (www.ncbi.nlm.nih.gov). The DNA copy numbers of the allelic alterations in cholangiocarcinoma were determined by quantitative real-time PCR and interpreted as allelic loss or DNA amplification by comparison with the reference gene. Associations between allelic imbalance and clinicopathological parameters of ICC patients were evaluated by χ2-test. The Kaplan-Meier method was used to analyze survival rates. RESULTS:From 33 primers,an altered DNA fragment (518 bp) amplified from BC17 random primer was found frequently in the tumors analyzed and mapped to chromosome 17p13.2. Sixteen of 52 (31%) cases showed DNA amplification,while 7 (13%) showed allelic loss. Interestingly,DNA amplification on chromosome 17p13.2 was associated with a good prognosis,median survival time (wk) of amp vs no amp was 44.14 vs 24.14,P=0.002; whereas allelic loss of this DNA sequence corresponded with a poor prognosis,median survival time (wk) of loss vs no loss was 18.00 vs 28.71,P=0.019). Moreover,Kaplan-Meier curves comparing the DNA alterations with survival depicted highly significant separation that the median survival time equal to DNAamplification,allelic loss,and normal was 44.14 wk,18.00 wk,and 24.29 wk,respectively (P=0.005). CONCLUSION:Alterations in the DNA sequence on chromosome 17p13.2 may be involved in cholangio-carcinogenesis,and could be used as a prognostic marker in the treatment of ICC patients.
AIM: To characterize and evaluate DNA alterations among intrahepatic cholangiocarcinoma (ICC) patients. METHODS: DNA from tumor and corresponding normal tissues of 52 patients was amplified with 33 arbitrary primers. The DNA fragment that alters most frequently in ICC was cloned, sequenced, and identified by comparison with known nucleotide sequences in the genome database (www.ncbi.nlm.nih.gov). The DNA copy numbers of the allelic alterations in cholangiocarcinoma were determined by quantitative real-time PCR and interpreted as allelic loss or DNA amplification by Associations between allelic imbalance and clinicopathological parameters of ICC patients were evaluated by χ2-test. The Kaplan-Meier method was used to analyze survival rates. RESULTS: From 33 primers, an altered DNA fragment (518 bp) from BC17 random primer was found frequently in the tumor analyzed and mapped to chromosome 17p13.2. Sixteen of 52 (31%) cases showed DNA amplificatio Interestingly, DNA amplification on chromosome 17p13.2 was associated with a good prognosis, median survival time (wk) of amp vs no amp was 44.14 vs 24.14, P = 0.002; loss of this DNA sequence corresponded with a poor prognosis, median survival time (wk) of loss vs no loss was 18.00 vs 28.71, P = 0.019). Moreover, Kaplan-Meier curves comparing the DNA alterations with survival rendered highly significant separation that the median survival time equal to DNA amplification, allelic loss, and normal was 44.14 wk, 18.00 wk, and 24.29 wk, respectively (P = 0.005). CONCLUSION: Alterations in the DNA sequence on chromosome 17p13.2 may be involved in cholangio-carcinogenesis, and could be used as a prognostic marker in the treatment of ICC patients.