膀胱移行上皮细胞癌患者染色体畸变与血清氧化应激状态的相关性
|
摘要
目的探讨3、7、17号染色体区带特异性探针(CSP3、CSP7、CSP17)和9号染色体p21(9p21)区p16基因位点特异性探针(GLPp16)的荧光原位杂交(fluorescence in situ hybridization,FISH)检测和血清氧化应激参数测定在膀胱移行上皮细胞癌(bladder transitional cell carcinoma,BTCC)中的应用价值,并分析其结果间的相互关系。方法随机选择健康对照30例和BTCC患者188例,运用FISH技术检测其膀胱脱落细胞的CSP3、CSP7、CSP17及GLPp16异常率,同时测定其血清TOS、TAS浓度和OSI值,比较各测定结果在健康人群和BTCC患者间的差异及其与BTCC分期分级的相关性,并进一步分析BTCC患者中CSP3、CSP7、CSP17和GLPp16异常分别和血清氧化应激参数的相关性。结果 BTCC患者与健康对照相比,其膀胱脱落细胞CSP3异常率(47.3%vs.10.0%)、CSP7异常率(52.1%vs.15.0%)、CSP17异常率(41.5%vs.10.0%)、GLPp16异常率(59.0%vs.20.0%)和FISH阳性率(50.5%vs.0.0%)以及其血清TOS浓度[(18.60±3.70).vs.(14.74±1.31)μmol H2O2 Eq./L]、OSI值[(1.33±0.41)vs.(0.89±0.10)]均明显增高(P<0.01),仅其血清TAS浓度[(1.44±0.21)vs.(1.66±0.16)mmol Trolox Eq./L]明显降低(P=0.000)。各探针异常率、FISH阳性率和血清氧化应激参数与BTCC分期之间均不相关(P>0.05)。而与BTCC分级之间,除GLPp16外,CSP3、CSP7、CSP17异常率和FISH阳性率呈明显的Gamma等级相关(r=-0.452、-0.564、-0.519和-0.375,P=0.000),且血清TOS、TAS浓度和OSI值也具有良好的Spearman相关性(r=0.678、-0.311和0.664,均P=0.000)。进一步分析4种染色体异常与3项氧化应激测定结果的相关性显示,CSP3、CSP7、CSP17异常率分别与TOS、OSI结果间均呈正相关(P<0.01),CSP7、CSP17异常率分别与TAS结果间呈负相关(P<0.01);而GLPp16异常率与TOS、TAS、OSI结果间均不相关(P>0.05)。结论 FISH检测膀胱脱落细胞染色体畸变是早期辅助诊断BTCC的有效的、无创性手段,而血清氧化应激参数准确定量可用于病情评估,二者的相关性研究有助于探索染色体畸变的机制以及其与BTCC发生、发展的关系。
Objective To investigate the application value of fluorescence in situ hybridization(FISH)detection for 3,7,17 chromosome region-specific probes(CSP3,CSP7,CSP17)and p16 gene locus-specific probe(GLPp16),and serum oxidative stress states determination in diagnosing bladder transitional cell carcinoma(BTCC),and analyze the interrelation of their results.Methods 30 health controls and 188 patients with BTCC were randomly enrolled to be detected the abnormal rate of CSP3,CSP7,CSP17 and GLPp16 in bladder exfoliated cells by FISH,and to be measured serum TOS,TAS concentrations and OSI values at the same time.Afterwards,their deference between healthy controls and patients with BTCC were compared,as well as their correlation with BTCC stages and classifications,and further analyzed the interrelation between serum oxidative stress parameters and the probe signal of CSP3,CSP7,CSP17 and GLPp16,respectively.Results In patients with BTCC comparing with healthy controls,the abnormal rate of CSP3(47.3% vs.10.0%),CSP7(52.1% vs.15.0%),CSP17(41.5% vs.10.0%),GLPp16(59.0% vs.20.0%),and FISH positive incidence(50.5%vs.0.0%)in bladder exfoliated cells were obviously increased(P<0.01),as well as serum TOS concentrations[(18.60±3.70)vs(14.74±1.31)μmol H2O2 Eq./L]and OSI values(1.33±0.41 vs.0.89±0.10),and only serum TAS concentrations[(1.44±0.21)vs.(1.66±0.16)mmol Trolox Eq./L]were obviously reduced(P=0.000).With BTCC stages,every chromosomal aberration rate,FISH positive incidence rate and serum oxidative stress parameters had no apparent correlations(P>0.05).But with BTCC classifications,except GLPp16,not only the aberration rate of CSP3,CSP7,CSP17,and FISH positive incidence had apparent Gamma rank correlations(r=-0.452,-0.564,-0.519,and-0.375,P=0.000),but also serum TOS,TAS concentrations and OSI values had superior Spearman correlation coefficients(r=0.678、-0.311和0.664,P=0.000).By further analyzing the correlation between four chromosomal aberration and three oxidative stress parameters results,it was shown that the abnormal rate of CSP3,CSP7,CSP17 had positive correlations with TOS and TAS results(P<0.01),moreover,the abnormal rate of CSP7 and CSP17 had negative correlations with TAS results(P<0.01),but GLPp16 abnormal rate had no correlations with TOS,TAS and OSI results(P>0.05).ConclusionThe chromosomal aberration detection by FISH in bladder exfoliated cells is an efficient and noninvasively means for early auxiliary diagnosing BTCC,and serum oxidative stress parameters determination may be helpful for disease assessment,the relationship study of their results should be contribute to exploring mechanisms of chromosomal aberration,and their relations with the genesis and development of BTCC.
Objective To investigate the application value of fluorescence in situ hybridization(FISH)detection for 3,7,17 chromosome region-specific probes(CSP3,CSP7,CSP17)and p16 gene locus-specific probe(GLPp16),and serum oxidative stress states determination in diagnosing bladder transitional cell carcinoma(BTCC),and analyze the interrelation of their results.Methods 30 health controls and 188 patients with BTCC were randomly enrolled to be detected the abnormal rate of CSP3,CSP7,CSP17 and GLPp16 in bladder exfoliated cells by FISH,and to be measured serum TOS,TAS concentrations and OSI values at the same time.Afterwards,their deference between healthy controls and patients with BTCC were compared,as well as their correlation with BTCC stages and classifications,and further analyzed the interrelation between serum oxidative stress parameters and the probe signal of CSP3,CSP7,CSP17 and GLPp16,respectively.Results In patients with BTCC comparing with healthy controls,the abnormal rate of CSP3(47.3% vs.10.0%),CSP7(52.1% vs.15.0%),CSP17(41.5% vs.10.0%),GLPp16(59.0% vs.20.0%),and FISH positive incidence(50.5%vs.0.0%)in bladder exfoliated cells were obviously increased(P<0.01),as well as serum TOS concentrations[(18.60±3.70)vs(14.74±1.31)μmol H2O2 Eq./L]and OSI values(1.33±0.41 vs.0.89±0.10),and only serum TAS concentrations[(1.44±0.21)vs.(1.66±0.16)mmol Trolox Eq./L]were obviously reduced(P=0.000).With BTCC stages,every chromosomal aberration rate,FISH positive incidence rate and serum oxidative stress parameters had no apparent correlations(P>0.05).But with BTCC classifications,except GLPp16,not only the aberration rate of CSP3,CSP7,CSP17,and FISH positive incidence had apparent Gamma rank correlations(r=-0.452,-0.564,-0.519,and-0.375,P=0.000),but also serum TOS,TAS concentrations and OSI values had superior Spearman correlation coefficients(r=0.678、-0.311和0.664,P=0.000).By further analyzing the correlation between four chromosomal aberration and three oxidative stress parameters results,it was shown that the abnormal rate of CSP3,CSP7,CSP17 had positive correlations with TOS and TAS results(P<0.01),moreover,the abnormal rate of CSP7 and CSP17 had negative correlations with TAS results(P<0.01),but GLPp16 abnormal rate had no correlations with TOS,TAS and OSI results(P>0.05).ConclusionThe chromosomal aberration detection by FISH in bladder exfoliated cells is an efficient and noninvasively means for early auxiliary diagnosing BTCC,and serum oxidative stress parameters determination may be helpful for disease assessment,the relationship study of their results should be contribute to exploring mechanisms of chromosomal aberration,and their relations with the genesis and development of BTCC.
引文
[1]Kanojia D,Garg M,Saini S,et al.Sperm Associated Antigen 9Plays an Important Role in Bladder Transitional Cell Carcinoma[J].PLoS One,2013,8(12):e81348.
[2]Siegel R,Ma J,Zou Z,Jemal A.Cancer statistics,2014[J].CA Cancer J Clin,2014,64(1):9-29.
[3]Feng JF,Lu L,Zeng P,et al.Serum total oxidant/antioxidant status and trace element levels in breast cancer patients[J].Int J Clin Oncol,2012,17(6):575-83.
[4]Wang D,Feng JF,Zeng P,et al.Total oxidant/antioxidant status in sera of patients with thyroid cancers[J].Endocr Relat Cancer,2011,18(6):773-782.
[5]Leufkens AM,Van Duijnhoven FJ,Woudt SH,et al.Biomarkers of oxidative stress and risk of developing colorectal cancer:a cohort-nested case-control study in the European Prospective Investigation Into Cancer and Nutrition[J].Am J Epidemiol,2012,175(7):653-663.
[6]Bubendorf L,Piaton E.UroVysion c multiprobe FISH in the triage of equivocal urinary cytology cases[J].Ann Pathol,2012,32(6):e52-56,438-443.
[7]Huang WT,Li LY,Pang J,et al.Fluorescence in situ hybridization assay detects upper urinary tract transitional cell carcinoma in patients with asymptomatic hematuria and negative urine cytology[J].Neoplasma,2012,59(4):355-360.
[8]Erel O.A new automated colorimetric method for measuring total oxidant status[J].Clin Biochem,2005,38(12):1103-1111.
[9]Erel O.A novel automated direct measurement method for total antioxidant capacity using a new generation,more stable ABTS radical cation[J].Clin Biochem,2004,37(4):277-285.
[10]Kwak KW,Kim SH,Lee HM.The utility of fluorescence in situ hybridization for detection of bladder urothelial carcinoma in routine clinical practice[J].J Korean Med Sci,2009,24(6):1139-1144.
[11]Caraway NP,Katz RL.A review on the current state of urine cytology emphasizing the role of fluorescence in situ hybridization as an adjunct to diagnosis[J].Cancer Cytopathol,2010,118(4):175-183.
[12]Sarosdy MF,Kahn PR,Ziffer MD,et al.Use of a multitarget fluorescence in situ hybridization assay to diagnose bladder cancer in patients with hematuria[J].J Urol,2006,176(1):44-47.
[13]Salim EI,Morimura K,Menesi A,et al.Elevated oxidative stress and DNA damage and repair levels in urinary bladder carcinomas associated with schistosomiasis[J].Int J Cancer,2008,123(3):601-608.
[14]Vadhanam MV,Thaiparambil J,Gairola CG,et al.Oxidative DNA adducts detected in vitro from redox activity of cigarette smoke constituents[J].Chem Res Toxicol,2012,25(11):2499-2504.
[15]Bellavia M,Gioviale MC,Damiano G,et al.Dissecting the different biological effects of oncogenic Ras isoforms in cancer cell lines:could stimulation of oxidative stress be the one more weapon of H-Ras?Regulation of oxidative stress and Ras biological effects[J].Med Hypotheses,2012,79(6):731-734.
[16]Guo S,Mao X,Chen J,et al.Overexpression of Pim-1in bladder cancer[J].J Exp Clin Cancer Res,2010,29:161-167.
[17]Sak SC,Barrett JH,Paul AB,et al.DNA repair gene XRCC1polymorphisms and bladder cancer risk[J].BMC Genet,2007,8:13-20.
[18]Stern MC,Lin J,Figueroa JD,et al.Polymorphisms in DNA repair genes,smoking,and bladder cancer risk:findings from the international consortium of bladder cancer[J].Cancer Res,2009,69(17):6857-6864.
[19]Wang Z,Rhee DB,Lu J,et al.Characterization of oxidative guanine damage and repair in mammalian telomeres[J].PLoS Genet,2010,6(5):e1000951-e1000963.
[20]Glei M,Hovhannisyan G,Pool-Zobel BL.Use of Comet-FISH in the study of DNA damage and repair:review[J].Mutat Res,2009,681(1):33-43.
[21]Jenkins NC,Liu T,Cassidy P,et al.The GLPp16(INK4A)tumor suppressor regulates cellular oxidative stress[J].Oncogene,2011,30(3):265-274.
[2]Siegel R,Ma J,Zou Z,Jemal A.Cancer statistics,2014[J].CA Cancer J Clin,2014,64(1):9-29.
[3]Feng JF,Lu L,Zeng P,et al.Serum total oxidant/antioxidant status and trace element levels in breast cancer patients[J].Int J Clin Oncol,2012,17(6):575-83.
[4]Wang D,Feng JF,Zeng P,et al.Total oxidant/antioxidant status in sera of patients with thyroid cancers[J].Endocr Relat Cancer,2011,18(6):773-782.
[5]Leufkens AM,Van Duijnhoven FJ,Woudt SH,et al.Biomarkers of oxidative stress and risk of developing colorectal cancer:a cohort-nested case-control study in the European Prospective Investigation Into Cancer and Nutrition[J].Am J Epidemiol,2012,175(7):653-663.
[6]Bubendorf L,Piaton E.UroVysion c multiprobe FISH in the triage of equivocal urinary cytology cases[J].Ann Pathol,2012,32(6):e52-56,438-443.
[7]Huang WT,Li LY,Pang J,et al.Fluorescence in situ hybridization assay detects upper urinary tract transitional cell carcinoma in patients with asymptomatic hematuria and negative urine cytology[J].Neoplasma,2012,59(4):355-360.
[8]Erel O.A new automated colorimetric method for measuring total oxidant status[J].Clin Biochem,2005,38(12):1103-1111.
[9]Erel O.A novel automated direct measurement method for total antioxidant capacity using a new generation,more stable ABTS radical cation[J].Clin Biochem,2004,37(4):277-285.
[10]Kwak KW,Kim SH,Lee HM.The utility of fluorescence in situ hybridization for detection of bladder urothelial carcinoma in routine clinical practice[J].J Korean Med Sci,2009,24(6):1139-1144.
[11]Caraway NP,Katz RL.A review on the current state of urine cytology emphasizing the role of fluorescence in situ hybridization as an adjunct to diagnosis[J].Cancer Cytopathol,2010,118(4):175-183.
[12]Sarosdy MF,Kahn PR,Ziffer MD,et al.Use of a multitarget fluorescence in situ hybridization assay to diagnose bladder cancer in patients with hematuria[J].J Urol,2006,176(1):44-47.
[13]Salim EI,Morimura K,Menesi A,et al.Elevated oxidative stress and DNA damage and repair levels in urinary bladder carcinomas associated with schistosomiasis[J].Int J Cancer,2008,123(3):601-608.
[14]Vadhanam MV,Thaiparambil J,Gairola CG,et al.Oxidative DNA adducts detected in vitro from redox activity of cigarette smoke constituents[J].Chem Res Toxicol,2012,25(11):2499-2504.
[15]Bellavia M,Gioviale MC,Damiano G,et al.Dissecting the different biological effects of oncogenic Ras isoforms in cancer cell lines:could stimulation of oxidative stress be the one more weapon of H-Ras?Regulation of oxidative stress and Ras biological effects[J].Med Hypotheses,2012,79(6):731-734.
[16]Guo S,Mao X,Chen J,et al.Overexpression of Pim-1in bladder cancer[J].J Exp Clin Cancer Res,2010,29:161-167.
[17]Sak SC,Barrett JH,Paul AB,et al.DNA repair gene XRCC1polymorphisms and bladder cancer risk[J].BMC Genet,2007,8:13-20.
[18]Stern MC,Lin J,Figueroa JD,et al.Polymorphisms in DNA repair genes,smoking,and bladder cancer risk:findings from the international consortium of bladder cancer[J].Cancer Res,2009,69(17):6857-6864.
[19]Wang Z,Rhee DB,Lu J,et al.Characterization of oxidative guanine damage and repair in mammalian telomeres[J].PLoS Genet,2010,6(5):e1000951-e1000963.
[20]Glei M,Hovhannisyan G,Pool-Zobel BL.Use of Comet-FISH in the study of DNA damage and repair:review[J].Mutat Res,2009,681(1):33-43.
[21]Jenkins NC,Liu T,Cassidy P,et al.The GLPp16(INK4A)tumor suppressor regulates cellular oxidative stress[J].Oncogene,2011,30(3):265-274.