胆管癌患者胆汁中端粒酶逆转录酶和Bcl-2基因的检测及意义
摘要
目的1.研究端粒酶逆转录酶(hTERT)和Bcl-2基因在胆管癌患者胆汁中的表达及相互关系。2.探讨检测胆汁中hTERT和Bcl-2基因对胆管癌的诊断价值。
方法1.对照组(胆总管结石患者胆汁标本20份),试验组(胆管癌患者胆汁标本20份),2.采用半定量逆转录一聚合酶链反应(RT-PCR)方法检测标本中hTERT和Bcl-2基因。
结果1.20例胆管癌患者胆汁中hTERT基因表达阳性14例(14/20),20例胆总管结石患者胆汁中hTERT基因无表达,两组间有统计学差异(p＜0.05);2.20例胆管癌患者胆汁中Bcl-2基因表达阳性11例(11/20),20例胆总管结石患者胆汁中Bcl-2基因的阳性1例(1/20),两组间有统计学差异(p＜0.05);3.10例胆管癌手术者中的6例有转移的胆管癌胆汁hTERT基因表达阳性6例(6/6)显著高于无转移的胆管癌胆汁hTERT基因表达(1/4)(p＜0.05)。10例胆管癌手术者中的4例高、中分化胆管癌中,其胆汁中Bcl-2基因阳性4例(4/4),低分化胆管癌患者胆汁中Bcl-2表达(2/6),两组无显著差异。4.胆管癌患者胆汁中hTERT和Bcl-2基因表达无相关性。
结论1.胆管癌患者胆汁hTERT基因与Bcl-2基因的表达均明显高于胆总管结石者(p＜0.05)。2.端粒酶和Bcl-2在胆管癌胆汁中的表达无相关性。3.用RT-PCR方法检测胆汁中hTERT和Bcl-2基因的表达对胆管癌的诊断有参考意义。
Objective:1.To study the expression of telomerase reverse transcriptase(hTERT) and Bcl-2 in bile of patients with cholangiocarcinoma and the relationship between them.2.To discuss the diagnostic value of the determination of the expression of hTERT and Bcl-2 in bile for cholangiocarcinoma.
Methods:1.The control group(20 bile samples of patients with choledocholith).The trial group(20 bile samples of patients with cholangiocarcinoma).2.Expression of hTERT and Bcl-2 in bile were confirmed by reverse transcriptase polymerase chain reaction(RT-PCR)
Results:1.hTERT were expressed in 14 out of the 20 bile samples of patients with cholangiocarcinoma(14/20),but no expression was found in 20 bile samples of patients with choledocholith,which was significantly different between the two groups(p＜0.05);2.Bcl-2 were expressed in 11 out of the 20 bile samples of patients with cholangiocarcinoma(11/20),but in only 1 out of the 20 bile samples of patients with choledocholith(1/20)、which was significantly different between the two groups(p＜0.05);3.In 10 patients with cholangiocarcinoma surgery, hTERT were expressed in 6 out of 6 bile samples of patients with metastatic cholangiocarcinoma(6/6),which was significantly higher than that of patients with non-metastatic cholangiocarcinoma(1/4)(p＜0.05).In 10 patients with cholangiocarcinoma surgery,Bcl-2 were expressed in 4 out of 4 bile samples of patients with highly or moderately differentiated cholangiocarcinoma(4/4),Bcl-2 were expressed in 2 out of 6 bile samples of patients with poorly differentiated cholangiocarcinoma(2/6),there no significant difference between them.4.The expression of hTERT and Bcl-2 in bile of patients with cholangiocarcinoma had no correlation.
Conclusion:1.The expression of hTERT and Bcl-2 in bile of patients with cholangiocarcinoma were significantly higher than those of patients with choledocholith(p＜0.05).2.The expression of hTERT and Bcl-2 in bile of patients with cholangiocarcinoma had no correlation.3.Determination of the expression of hTERT and Bcl-2 in bile by RT-PCR has diagnostic value for cholangiocarcinoma.
方法1.对照组(胆总管结石患者胆汁标本20份),试验组(胆管癌患者胆汁标本20份),2.采用半定量逆转录一聚合酶链反应(RT-PCR)方法检测标本中hTERT和Bcl-2基因。
结果1.20例胆管癌患者胆汁中hTERT基因表达阳性14例(14/20),20例胆总管结石患者胆汁中hTERT基因无表达,两组间有统计学差异(p＜0.05);2.20例胆管癌患者胆汁中Bcl-2基因表达阳性11例(11/20),20例胆总管结石患者胆汁中Bcl-2基因的阳性1例(1/20),两组间有统计学差异(p＜0.05);3.10例胆管癌手术者中的6例有转移的胆管癌胆汁hTERT基因表达阳性6例(6/6)显著高于无转移的胆管癌胆汁hTERT基因表达(1/4)(p＜0.05)。10例胆管癌手术者中的4例高、中分化胆管癌中,其胆汁中Bcl-2基因阳性4例(4/4),低分化胆管癌患者胆汁中Bcl-2表达(2/6),两组无显著差异。4.胆管癌患者胆汁中hTERT和Bcl-2基因表达无相关性。
结论1.胆管癌患者胆汁hTERT基因与Bcl-2基因的表达均明显高于胆总管结石者(p＜0.05)。2.端粒酶和Bcl-2在胆管癌胆汁中的表达无相关性。3.用RT-PCR方法检测胆汁中hTERT和Bcl-2基因的表达对胆管癌的诊断有参考意义。
Objective:1.To study the expression of telomerase reverse transcriptase(hTERT) and Bcl-2 in bile of patients with cholangiocarcinoma and the relationship between them.2.To discuss the diagnostic value of the determination of the expression of hTERT and Bcl-2 in bile for cholangiocarcinoma.
Methods:1.The control group(20 bile samples of patients with choledocholith).The trial group(20 bile samples of patients with cholangiocarcinoma).2.Expression of hTERT and Bcl-2 in bile were confirmed by reverse transcriptase polymerase chain reaction(RT-PCR)
Results:1.hTERT were expressed in 14 out of the 20 bile samples of patients with cholangiocarcinoma(14/20),but no expression was found in 20 bile samples of patients with choledocholith,which was significantly different between the two groups(p＜0.05);2.Bcl-2 were expressed in 11 out of the 20 bile samples of patients with cholangiocarcinoma(11/20),but in only 1 out of the 20 bile samples of patients with choledocholith(1/20)、which was significantly different between the two groups(p＜0.05);3.In 10 patients with cholangiocarcinoma surgery, hTERT were expressed in 6 out of 6 bile samples of patients with metastatic cholangiocarcinoma(6/6),which was significantly higher than that of patients with non-metastatic cholangiocarcinoma(1/4)(p＜0.05).In 10 patients with cholangiocarcinoma surgery,Bcl-2 were expressed in 4 out of 4 bile samples of patients with highly or moderately differentiated cholangiocarcinoma(4/4),Bcl-2 were expressed in 2 out of 6 bile samples of patients with poorly differentiated cholangiocarcinoma(2/6),there no significant difference between them.4.The expression of hTERT and Bcl-2 in bile of patients with cholangiocarcinoma had no correlation.
Conclusion:1.The expression of hTERT and Bcl-2 in bile of patients with cholangiocarcinoma were significantly higher than those of patients with choledocholith(p＜0.05).2.The expression of hTERT and Bcl-2 in bile of patients with cholangiocarcinoma had no correlation.3.Determination of the expression of hTERT and Bcl-2 in bile by RT-PCR has diagnostic value for cholangiocarcinoma.
引文
[1]Seyama Y,Kubota K,Sano K,et al.Long-term outcome of extended hemihepatectomy for hilar bile duct cancer with nomortality and high survival rate.Ann Surg,2003,238(1):73-83.
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[4]王征旭,何振平,房殿春,等.胆管癌K-ras癌基因,p53抑癌基因突变及表达的研究.癌症,1997,16(1):48.
[5]Chow NH,Huang SM,Chan SH,et al.Significance of c-erB 2 expression in normal and neoplastic epithelium of biliary tract.Anticancer Res,1995,15(3):1055.
[6]Voravud N,Foster CS,Gilbertson JA,et al.Oncogene expression in cholangiocarcinoma and in normal hepatic development.Hum Pathol,1989,20(12):1163.
[7]Kin NW,Piatyszek MA,Prowse KK,et al.Specific association of human telomerase activity with immortal cells and cancer.Science,1994,(26)6:2011.
[8]Tahara H,Nakanish T,Kitamato M,et al.Telomerase activity in human liver tissues.Comparison between chronic liver disease and hepatocellular carcinomas.Cancer Res,1995,55:27-35.
[9]Cohn M,Blackburn EH.Telomerase in yeast.Science,1995,26(9):396.
[10]Hiyoma E,Kodama T,Shinbara K,er al.Telomerase activity is detected in pancreatic cancer but in benign tumors.Cancer Res,1997,57:326.
[11]Mokbel K.The role of telomerase in breast cancer.Eur J Surg Oncol,2000,26:509.
[12]Okaro AC,Decery AR,Hutchins RR,et al.The expression of antiapoptotic proteins Bcl-XL and Mcl-1 in benign,dysplastic and malignant biliary epithelium.Clinical pathology,2002,54:927-932.
[13]Lyonel G,Esther D,Apoptosis J.The oncologist.1999,4(4):332-339.
[14]Holzinger F,ZI Graggen K,Buchler MW.Mechanism of biliary carcinogenesis:a pathogentic multi-stage cascade towards cholangiocarcinoma.Ann Oncol,1999,10(supps14):122-126.
[15]Ortiz-Rey J,Alvarez C,Anton-Badiola I,et al.Human meissner corpuseles express Bcl-2 but not Bax protein.Neruosci Lett,2002,239(2):240.
[16]Bukholm IR,Bukholm G,Nesland JM.Reduced expression of both Bax and Bcl-2 is independently associated with lymph node metastasis in human breast carcinomas.APMIS,2002,110(3):214-220.
[17]Kubicka S,Kuhnel F,Flemming P,et al.K-ras mutations in the bile of patients with primary sclerosing cholangitis.Gut,2001,48:403-408.
[18]塔西乌,王炳生,刘银昆,等.检测胆汁和胆管刷取物中K-ras基因突变诊断胆管 癌.中华试验外科杂志,2001,21(3):319-320.
[19]李兆申,刘枫,周国雄,等.胰液中肿瘤相关基因检测在胰腺癌诊断中的价值.中华医学杂志,2004,84(24):2091-2093.
[20]Mandal M,Kumar R.Bcl-2 modulates telomerase activity.J Biol Chem 1997,272(22):14183-14187.
[21]郭华雄,余龙江,周治兰,等.乳腺癌端粒酶活性与Bcl-2和Bax蛋白表达的相关性研究.肿瘤防治研究,2002,29(2):112-113.
[22]熊小熊,陈蔚蔚,周宏远,等.端粒酶基因和抗凋亡基因Bcl-2在肺组织中的表达研究.肿瘤防治研究,2002,29(3):165-167.
[23]Kin NW,Piatyszek MA,Prow se KK,et al.Specific association If human telomerase activity with immortal cells and cancer.Science,1994,266:2011.
[24]Tahara H,Nakanish T,Kitamato M,et al.Telomerase activity in human liver tissues comparison between chronic liver disease and hepatocellular carcinomas.Cancer Res,1995,55:2735.
[25]Cohn M,Blackbum EH.Telomerase in yeast.Science,1995,269:396.
[26]Hiyama E,Yokoyama T,Tstsnnoto N,et al.Telomerase activity in gastric cancer.Cancer Res,1995,55(15):3258-3262.
[27]Rhyu M S.Telomerase and immortality.J Natl Cancer Inst,1995,87(12):884-894.
[28]Yoshida K,Sugino T,Tshara H,et al.Telomerase activity in bladder carcinoma and its implication for noninvasive diagnosis by detection of exfoliated cancer cells in urine.Cancer,1997,79(2):362-369.
[29]Bieche I,Nogues C,Paradis V,et al.Quantitation of hTERT gene expression in sporadic breast tumors with a real-time reverse transcription-polymerase chain reaction assay.Clin Cancer Res,2000,6:452.
[30]Longchampt E,Lebret T,Molinie V,et al.Detection of telomerase status by semi-quantitative and in situ assays,and by real-time reverse transcription-polymerase chain reaction(telomerase reverse transcriptase)assay in bladder carcinomas.BJU Int,2003,91(6):567.
[31]Lledo SM,Garcia-Granero E,Dasi F,et al.Real time quantification in plasma of human telomerase reverse transcriptase(hTERT)mRNA in patients with colorectal cancer.Colorectal Dis,2004,6(4):236.
[32]Niiyama H,Mizumoto K,Kusumoto M,et al.Activity of telomerase and diagnostic application in biopsy specimens from biliary tract neoplasms.Cancer,1999,85(10):2138-2143.
[33]Virkajarvi N,Pakko P,Soini Y.Apoptotic index and apoptosis influencing proteins Bcl-2,mcl-2,Bax and caspases 3,6 and 8 in pancreatic carcinoma J.Histopthology,1998,33:432-439.
[34]Wu X,Blank A,Olovsson M,et al.Expression fo Bcl-2,Bcl-x,Mcl-1,Bax and Bak in human uterine leiomyomas and myometrium during the menstrual cycle and after menopause J.J Steroid Biochem Mol Biol,2002,81(1):77-83.
[35]王徽,宣兆艳,刘永晨,等。Bcl-2基因在乳腺癌中的表达及临床意义.中国癌症杂志,2001,11(3):252-253.
[36]Ito Y,Takeda T,Sasaki Y,et al.Bcl-2 expression in cholangiocellular carcinoma is inversely correlated with biological aggressive phenotypes.J Oncology,2000,59(1):63-67.
[37]Charlotte F,L'Hermine A,Martin N,et al.Immunohistochemical detection of Bcl-2 protein in normal and pathological human liver.Am J Pathol,1994,144(3):460-465.
[38]Ito Y,Takeda T,Sasaki Y,et al.Expression and clinical implication of Bcl-2 in extrahepatic bile duct carcinoma:its relationship with biological features.J Anticancer Res,2000,20(5c):3891-3895.
[39]Harnois DM,Que FG,Celli A,et al.Bcl-2 is overe expressed and alters the threshold for apoptosis in a cholangiocarinoma cell line.J Hepatol,1997,26(4):884-890.
[40]Mandal M,Kumar R.Bcl-2 modulates telomerase activity.J Biol Chem,1997,272(22):14183-14187.
[1]Kubicka S,Kuhnel F,Flemming P,et al.K-ras mutations in the bile of patients with primary sclerosing cholangitis.Gut,2001,48:403-408.
[2]Tsai CC,MoLR,Chou CY,et al Percutaneous transhepatic transluminal forceps biopsy in obstructive jaundice.Hepatogastroenterology,1997,44(15):770-773.
[3]Pugliese V,Conio M,Nicolo G,et al Endoscopic retrograde forceps biopsy and brush cytology of biliary strictures:a prospective study.Gastuointest Endosc,1995,42(6):520-526.
[4]Ponchon T,Gagnon P,Berger F et al Value of endobiliary brush cytology and biopsies for the diagnosis of malignant bile buct stenosis:results of a prospective study.1995-42(6):565-572.
[5]Bjornsson E,Kilander A,Olsson R,er al CA19-9 and CEA are unrealiable markers for chlolangiocarcinoma.Liver,1999,19:501-503.
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[11]塔西乌,王炳生,等.检测胆汁和胆管刷取物中K2ras基因突变诊断胆管癌.中华实验外科杂志,2004,21(3):319-320.
[12]俞世安,彭乘宏,吴荣进,等.胆管癌胆汁脱落细胞端粒酶活性检测的诊断价值.中华肝胆外科杂志,2003,9(8):469-470.
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[15]何贵金,杜怀林,夏振龙,等.胆管癌患者血清IL26检测意义.中国医科大学学报,2001,30(2):145-146.
[2]Holzinger E,ZI Graggen K,Buchler MW.Mechanism of biliary carcinogenesis:a pathogenetic muli-stagecascade towards cholangiocarcinoma.Ann oncol,1999,10(suppl4):122-126.
[3]Lee JC,Lin PW,Lin YJ,et al.Analysis of k-ras gene mutations in periampullary cancers,gallbladder cancers and cholangioncarcinomas from paraffin-embedded tissue sections.Formos Med Assoc,195,94(12):719.
[4]王征旭,何振平,房殿春,等.胆管癌K-ras癌基因,p53抑癌基因突变及表达的研究.癌症,1997,16(1):48.
[5]Chow NH,Huang SM,Chan SH,et al.Significance of c-erB 2 expression in normal and neoplastic epithelium of biliary tract.Anticancer Res,1995,15(3):1055.
[6]Voravud N,Foster CS,Gilbertson JA,et al.Oncogene expression in cholangiocarcinoma and in normal hepatic development.Hum Pathol,1989,20(12):1163.
[7]Kin NW,Piatyszek MA,Prowse KK,et al.Specific association of human telomerase activity with immortal cells and cancer.Science,1994,(26)6:2011.
[8]Tahara H,Nakanish T,Kitamato M,et al.Telomerase activity in human liver tissues.Comparison between chronic liver disease and hepatocellular carcinomas.Cancer Res,1995,55:27-35.
[9]Cohn M,Blackburn EH.Telomerase in yeast.Science,1995,26(9):396.
[10]Hiyoma E,Kodama T,Shinbara K,er al.Telomerase activity is detected in pancreatic cancer but in benign tumors.Cancer Res,1997,57:326.
[11]Mokbel K.The role of telomerase in breast cancer.Eur J Surg Oncol,2000,26:509.
[12]Okaro AC,Decery AR,Hutchins RR,et al.The expression of antiapoptotic proteins Bcl-XL and Mcl-1 in benign,dysplastic and malignant biliary epithelium.Clinical pathology,2002,54:927-932.
[13]Lyonel G,Esther D,Apoptosis J.The oncologist.1999,4(4):332-339.
[14]Holzinger F,ZI Graggen K,Buchler MW.Mechanism of biliary carcinogenesis:a pathogentic multi-stage cascade towards cholangiocarcinoma.Ann Oncol,1999,10(supps14):122-126.
[15]Ortiz-Rey J,Alvarez C,Anton-Badiola I,et al.Human meissner corpuseles express Bcl-2 but not Bax protein.Neruosci Lett,2002,239(2):240.
[16]Bukholm IR,Bukholm G,Nesland JM.Reduced expression of both Bax and Bcl-2 is independently associated with lymph node metastasis in human breast carcinomas.APMIS,2002,110(3):214-220.
[17]Kubicka S,Kuhnel F,Flemming P,et al.K-ras mutations in the bile of patients with primary sclerosing cholangitis.Gut,2001,48:403-408.
[18]塔西乌,王炳生,刘银昆,等.检测胆汁和胆管刷取物中K-ras基因突变诊断胆管 癌.中华试验外科杂志,2001,21(3):319-320.
[19]李兆申,刘枫,周国雄,等.胰液中肿瘤相关基因检测在胰腺癌诊断中的价值.中华医学杂志,2004,84(24):2091-2093.
[20]Mandal M,Kumar R.Bcl-2 modulates telomerase activity.J Biol Chem 1997,272(22):14183-14187.
[21]郭华雄,余龙江,周治兰,等.乳腺癌端粒酶活性与Bcl-2和Bax蛋白表达的相关性研究.肿瘤防治研究,2002,29(2):112-113.
[22]熊小熊,陈蔚蔚,周宏远,等.端粒酶基因和抗凋亡基因Bcl-2在肺组织中的表达研究.肿瘤防治研究,2002,29(3):165-167.
[23]Kin NW,Piatyszek MA,Prow se KK,et al.Specific association If human telomerase activity with immortal cells and cancer.Science,1994,266:2011.
[24]Tahara H,Nakanish T,Kitamato M,et al.Telomerase activity in human liver tissues comparison between chronic liver disease and hepatocellular carcinomas.Cancer Res,1995,55:2735.
[25]Cohn M,Blackbum EH.Telomerase in yeast.Science,1995,269:396.
[26]Hiyama E,Yokoyama T,Tstsnnoto N,et al.Telomerase activity in gastric cancer.Cancer Res,1995,55(15):3258-3262.
[27]Rhyu M S.Telomerase and immortality.J Natl Cancer Inst,1995,87(12):884-894.
[28]Yoshida K,Sugino T,Tshara H,et al.Telomerase activity in bladder carcinoma and its implication for noninvasive diagnosis by detection of exfoliated cancer cells in urine.Cancer,1997,79(2):362-369.
[29]Bieche I,Nogues C,Paradis V,et al.Quantitation of hTERT gene expression in sporadic breast tumors with a real-time reverse transcription-polymerase chain reaction assay.Clin Cancer Res,2000,6:452.
[30]Longchampt E,Lebret T,Molinie V,et al.Detection of telomerase status by semi-quantitative and in situ assays,and by real-time reverse transcription-polymerase chain reaction(telomerase reverse transcriptase)assay in bladder carcinomas.BJU Int,2003,91(6):567.
[31]Lledo SM,Garcia-Granero E,Dasi F,et al.Real time quantification in plasma of human telomerase reverse transcriptase(hTERT)mRNA in patients with colorectal cancer.Colorectal Dis,2004,6(4):236.
[32]Niiyama H,Mizumoto K,Kusumoto M,et al.Activity of telomerase and diagnostic application in biopsy specimens from biliary tract neoplasms.Cancer,1999,85(10):2138-2143.
[33]Virkajarvi N,Pakko P,Soini Y.Apoptotic index and apoptosis influencing proteins Bcl-2,mcl-2,Bax and caspases 3,6 and 8 in pancreatic carcinoma J.Histopthology,1998,33:432-439.
[34]Wu X,Blank A,Olovsson M,et al.Expression fo Bcl-2,Bcl-x,Mcl-1,Bax and Bak in human uterine leiomyomas and myometrium during the menstrual cycle and after menopause J.J Steroid Biochem Mol Biol,2002,81(1):77-83.
[35]王徽,宣兆艳,刘永晨,等。Bcl-2基因在乳腺癌中的表达及临床意义.中国癌症杂志,2001,11(3):252-253.
[36]Ito Y,Takeda T,Sasaki Y,et al.Bcl-2 expression in cholangiocellular carcinoma is inversely correlated with biological aggressive phenotypes.J Oncology,2000,59(1):63-67.
[37]Charlotte F,L'Hermine A,Martin N,et al.Immunohistochemical detection of Bcl-2 protein in normal and pathological human liver.Am J Pathol,1994,144(3):460-465.
[38]Ito Y,Takeda T,Sasaki Y,et al.Expression and clinical implication of Bcl-2 in extrahepatic bile duct carcinoma:its relationship with biological features.J Anticancer Res,2000,20(5c):3891-3895.
[39]Harnois DM,Que FG,Celli A,et al.Bcl-2 is overe expressed and alters the threshold for apoptosis in a cholangiocarinoma cell line.J Hepatol,1997,26(4):884-890.
[40]Mandal M,Kumar R.Bcl-2 modulates telomerase activity.J Biol Chem,1997,272(22):14183-14187.
[1]Kubicka S,Kuhnel F,Flemming P,et al.K-ras mutations in the bile of patients with primary sclerosing cholangitis.Gut,2001,48:403-408.
[2]Tsai CC,MoLR,Chou CY,et al Percutaneous transhepatic transluminal forceps biopsy in obstructive jaundice.Hepatogastroenterology,1997,44(15):770-773.
[3]Pugliese V,Conio M,Nicolo G,et al Endoscopic retrograde forceps biopsy and brush cytology of biliary strictures:a prospective study.Gastuointest Endosc,1995,42(6):520-526.
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