CAJ | 학술논문

目的找寻胆管癌细胞标记染色体及外周血标志物,为胆管癌临床的早期诊断提供依据.方法用染色体显带分析技术和荧光原位杂交(FISH)分析技术分析胆管癌组织、胆管癌细胞株染色体畸变及胆管癌患者外周血淋巴细胞染色体不稳定性和脆性部位的表达;应用聚合酶链反应-单链构象多态性(PCR-SSCP)及免疫组织化学技术(ABC)分析肝门部胆管癌的p16基因的突变及蛋白表达.结果瘤细胞染色体数目平均62.3条,以亚三倍体和超二倍体多见,染色体数目畸变率达83.67％;细胞株比新鲜瘤组织染色体数目变化范围大,且高倍体细胞相对较多.染色体结构畸变率为62.8％,其中9p高频率缺失(50.8％).p16基因变异率为58％(21/36),p16蛋白表达下降率共占50％(18/36).胆管癌患者外周血淋巴细胞染色体裂隙、断裂和异常细胞数目及细胞脆性部位表达率显著高于正常对照组.结论胆管癌发病与染色体畸变及癌基因激活或抑癌基因失活有关;胆管癌患者外周血淋巴细胞染色体裂隙、断裂和异常细胞比率明显增高,脆性部位表达明显异常.胆管癌外周血淋巴细胞染色体断裂点8p22、9p21、11q13、17p12出现率较高,是胆管癌相关断裂点,为寻找胆管癌外周血标志物提供候选部位.
목적 조심담관암세포표기염색체급외주혈표지물,위담관암림상적조기진단제공의거.방법 용염색체현대분석기술화형광원위잡교(FISH)분석기술분석담관암조직、담관암세포주염색체기변급담관암환자외주혈림파세포염색체불은정성화취성부위적표체;응용취합매련반응-단련구상다태성(PCR-SSCP)급면역조직화학기술(ABC)분석간문부담관암적p16기인적돌변급단백표체.결과 류세포염색체수목평균62.3조,이아삼배체화초이배체다견,염색체수목기변솔체83.67％;세포주비신선류조직염색체수목변화범위대,차고배체세포상대교다.염색체결구기변솔위62.8％,기중9p고빈솔결실(50.8％).p16기인변이솔위58％(21/36),p16단백표체하강솔공점50％(18/36).담관암환자외주혈림파세포염색체렬극、단렬화이상세포수목급세포취성부위표체솔현저고우정상대조조.결론 담관암발병여염색체기변급암기인격활혹억암기인실활유관;담관암환자외주혈림파세포염색체렬극、단렬화이상세포비솔명현증고,취성부위표체명현이상.담관암외주혈림파세포염색체단렬점8p22、9p21、11q13、17p12출현솔교고,시담관암상관단렬점,위심조담관암외주혈표지물제공후선부위.
Objective To study alterations of molecular cytogenetics of cholangiocarcinoma on chromosome and gene,study the possible involvement of chromosome abnormalities and the genetic susceptibility in pathogenesis of cholangiocarcinoma,investigate chromosome instability and expression of fragile sites of cholangiocarcinoma and relationship each other,and find marker chromosome of cholangiocarcinoma.Methods Chromosomal analysis and fluorescence in situ hybridization (FISH) were used with 9th chromosome libraries to investigate the chromosomal aberration,the chromosome instability and expressiom of fragile sites in patients with cholangiocarcinoma.The genetic alteration and expression of p16 gene were analyzed by polymerase chain reaction single strand comformation polymorphism (PCR-SSCP) and immunohistochemistry.Results Chromosomal aberrations were found in 10 case of patients and 1 case of cell line of cholangiocarcinoma.The chromosome model number was 62.3 and predominantly hyperdiploid,with a chromosome number aberration rate of 83.67％,and a structure aberration rate of 62.8％.9p deletion appeared more frequently(50.8％).p16 gene mutation rate was 58.3％ (21/36).The expression of p16 gene was not significantly related with sex and age of the patients,and had a relation with Bismuth type,the depth of invasion and lymph node metastasis.The chromosome gaps (8.75 ± 3.30),chromosome breakpoints (7.63 ± 2.76),abnormal cell rate (13.35 ± 4.73) and expression rate of fragile sites of the patients were markedly higher in patients with cholgnagiocarcinoma than those of normal persons (3.17 ±1.82,2.04±1.76,3.65 ±1.97) (P＜0.01).Conclusion Chromosomal aberrations and gene mutation may play an important role in the pathogenesis of cholangiocarcinoma.The alteration of p16 gene and abnormal expression of p16 protein are significantly correlated with the biological behaviors and clinical staging of cholangiocarcinorma and may hence be helpful to prognosis.The increase of chromosome instability and overexpression of frigile sites freguently exist in cholangiocarcinoma.The sites on 8p22,9p21,13q13 and 17p12 may be candidate regions for further identifying the blood marker chromosome of cholangiocarcinoma.