CAJ | 학술논문

目的分析血清及肝组织miR-200a在肝癌发生过程中的表达水平并将其与传统肝癌血清学标志物甲胎蛋白比较,探索其成为肝癌早期诊断标志物的可能性.方法二乙基亚硝胺腹腔注射诱导建立大鼠肝癌模型,建模过程中收集肝癌发展到各个阶段大鼠血液及肝脏组织.同时,临床收集正常、肝硬化、肝癌患者血液标本.用实时定量PCR技术检测血清miR-200a的相对表达情况,用酶联免疫吸附法检测血清甲胎蛋白动态变化,原位杂交法检测miR-200a在组织中表达情况.体质量数据采用两样本t检验,PCR及酶联免疫吸咐各组数据间采用单因素方差分析进行比较.结果与正常组及纤维化期大鼠相比,miR-200a在肝硬化期、肝癌早期、肝癌晚期的大鼠血清均有不同程度下调(P值均＜0.05),甲胎蛋白在肝癌早期、肝癌晚期的大鼠血清明显上调(P值均＜ 0.05).与肝硬化期大鼠比较,肝癌晚期大鼠体内的miR-200a表达显著下调(P值均＜0.05),甲胎蛋白在肝癌早期、肝癌晚期均显著上调.临床标本分析结果显示,miR-200a在肝硬化及肝癌患者中均显著下调(P值均＜0.05),甲胎蛋白仅在肝癌患者中出现异常表达.原位杂交结果显示,以正常肝组为标准,miR-200a在正常组、纤维化期、肝硬化期、肝癌早期、肝癌晚期中表达水平依次为:1.00％±0.01％、0.37％±0.03％、0.14％±0.01％、0.05％±0％、0.01％±0.00％.结论 miR-200a极大程度地参与了肝癌发生,对于肝癌发生各个阶段均有巨大的指示作用,可能作为肝癌预防和早期诊断的一个潜在标志物.
목적 분석혈청급간조직miR-200a재간암발생과정중적표체수평병장기여전통간암혈청학표지물갑태단백비교,탐색기성위간암조기진단표지물적가능성.방법 이을기아초알복강주사유도건립대서간암모형,건모과정중수집간암발전도각개계단대서혈액급간장조직.동시,림상수집정상、간경화、간암환자혈액표본.용실시정량PCR기술검측혈청miR-200a적상대표체정황,용매련면역흡부법검측혈청갑태단백동태변화,원위잡교법검측miR-200a재조직중표체정황.체질량수거채용량양본t검험,PCR급매련면역흡부각조수거간채용단인소방차분석진행비교.결과 여정상조급섬유화기대서상비,miR-200a재간경화기、간암조기、간암만기적대서혈청균유불동정도하조(P치균＜0.05),갑태단백재간암조기、간암만기적대서혈청명현상조(P치균＜ 0.05).여간경화기대서비교,간암만기대서체내적miR-200a표체현저하조(P치균＜0.05),갑태단백재간암조기、간암만기균현저상조.림상표본분석결과현시,miR-200a재간경화급간암환자중균현저하조(P치균＜0.05),갑태단백부재간암환자중출현이상표체.원위잡교결과현시,이정상간조위표준,miR-200a재정상조、섬유화기、간경화기、간암조기、간암만기중표체수평의차위:1.00％±0.01％、0.37％±0.03％、0.14％±0.01％、0.05％±0％、0.01％±0.00％.결론 miR-200a겁대정도지삼여료간암발생,대우간암발생각개계단균유거대적지시작용,가능작위간암예방화조기진단적일개잠재표지물.
Objective To explore whether microRNA-200a (miR-200a) could be used as a novel biomarker of liver cancer using a rat model system.Methods Diethylnirtosamine abdominal injection was applied to induce liver cancer in the F344 rat strain (n =40); ten unmodeled rats served as controls.In addition,human subjects with normal healthy liver (n =10),liver cirrhosis (n =10),and liver cancer (n =10) were enrolled in the study.Blood samples from both rats and patients and rats' livers were collected for analysis.Real-time quantitative PCR and enzyme-linked immunosorbent assay were used respectively to measure the expressions of serum miR-200a and alpha-fetoprotein (AFP) for all rat and human subjects.In situ hybridization was used to detect the miR-200a expression in the rats' livers.Results Comparison of normal rats and the liver cancer modeled rats showed that the latter had significantly lower expression of miR-200a (P ＜ 0.05),with decreasing expression following the progression of liver injury to cancer (liver cirrhosis rats ＜ early liver cancer rats ＜ advanced liver cancer rats); in contrast,the AFP levels were significantly higher in the liver cancer modeled rats only at the early and advanced stages of the liver cancer (P ＜ 0.05).These results suggested that miR-200a expression decreases during the developmental process of liver cancer,while AFP expression increases distinctly at the stage of tumor formation.Analysis of the human subjects' clinical samples showed that miR-200a expression was decreased in both liver cirrhosis patients and liver cancer patients (vs.normal liver subjects,P ＜ 0.05),while AFP showed abnormal expression only in the patients with liver cancer.Comparison of the normal rats and modeled rats using in situ hybridization showed the positive rates for miR-200a expression were 1.00％ ± 0.01％ in rats with normal liver,0.37％ ± 0.03％ in rats with fibrotic liver,0.14％ ± 0.01％ in rats with cirrhotic liver,0.05％ ± 0.00％ in rats with early stage liver cancer,and 0.01％ ± 0.00％ in rats with advanced stage liver cancer.Conclusion MiR-200a may play an important role in liver cancer development and may have diagnostic value for indicating early liver cancer.