高尔基体蛋白73及其基因检测对原发性肝癌诊断的价值

Diagnostic role of Golgi protein 73 and its gene determination in primary hepatic carcinoma

目的 评价分析GP73和GP73 mRNA在PHC中的诊断价值,探讨血清中GP73和AFP联合检测对PHC诊断和高危人群普查的意义,为PHC诊断和普查提供一种新方法 .方法 采用ELISA对73例PHC、13例肝硬化、32例肝炎和62名健康人的血清GP73、AFP水平进行检测,采用SYBR Green实时荧光定量PCR法检测各组外周血单个核细胞GP73 mRNA的相对表达量,以Ct值比较法计算GP73 mRNA相对表达水平,同时检测分析8份正常肝组织和8份肝癌组织的GP73 mRNA相对表达水平.结果 ELISA检测4组血清GF73、AFP结果 显示,总体比较经Kruskal-Wallis检验,4组间差异有统计学意义（H值分别为89.6、52.0,P均＜0.01）,全血GP73 mRNA含量4组间差异无统计学意义（H=4.33,P＞0.05）.组间多重比较Mann-Whitney检验结果显示,PHC组血清GP73的含量[166.7（162.7-231.8）μL]与肝硬化[57.3（46.6～113.6）μg/L]、肝炎[29.6（26.2～54.5）μg/L]及健康对照组[25.1（20.8～29.4）μg/L]比较,差异有统计学意义（U值分别为246、297、349,P均＜0.01）,各组血清AFP的含量分别为380.9（258.5～ 503.2）μg/L、3.8（1.3～14.5）μg/L、5.1（2.4～7.8）μg/L、2.8（2.2～5.7）μg/L,差异亦有统计学意义（U值分别为246、419、790,P均＜0.01）.肝癌组织GP73 mRNA表达量（12.64）显著高于正常肝组织（1.00）.以ROC曲线确定诊断PHC的GP73临界值为123.2μg/L和AFP临界值为10.6 μg/L时,PHC组血清GP73、AFP单项检测的敏感度分别为65.8%和53.4%,特异度分别为95.3%和92.5%,两者联合检测的敏感度为79.5%,特异度为90.7%.结论 GP73蛋白对PHC诊断具有较好的敏感度和特异度;全血标本GP73 mRNA检测不能作为诊断PHC的肿瘤标志,肝组织标本GP73 mRNA检测可作为诊断PHC的肿瘤标志,但存在创伤性大、风险大、患者痛苦等缺点.血清GP73联合AFP检测可有效提高PHC诊断,可用于PHC高危人群的普查及筛选.

Objective To explore the diagnostic value of GP-/3 protein in gene detection in the patient of primary hepatic carcinoma, to discuss the joint roles of serum GP73 and AFP, and provide a novel method for the diagnosis for PHC and screening for high-risk population. Methods ELISA was used to detect the serum level of GP73 and AFP in 73 cases of PHC, 13 cases of hepatic cirrhosis, 32 cases of hepatitis and 62 cases of health people. SYBR Green real time fluorescence quantitative PCR was used to detect the relative value of GP73 mRNA in the peripheral blood cells of each group. Comparative Ct method was used to evaluate the relative expression levels. Eight cases of normal liver tissues and 8 cases of PHC tissues were detected at the same time to compare the relative expression levels. Results Kruskal-Wallis test showed that the serum levels of GP73 and AFP had significant differences between four groups（H value were 89. 6 and 52.0, P ＜ 0. 01） and the whole blood GP73 mRNA had no significant differences（H =4. 33, P ＞ 0. 05）. Mann-Whitney test showed that the serum levels of GP73 had significant differences among PHC groups[166. 7 （162. 7-231.8） μg/L] and liver cirrhosis[57. 3 （46. 6-113. 6） μg/L], hepatitis[29. 6（26. 2-54. 5） μg/L], health group[25.1 （20. 8-29. 4） μg/L] （U value were 246, 297, 349, P ＜ 0. 01）.The A FP levels of the four groups were 380. 9 （258.5-503.2） μg/L, 3.8 （1.3-14. 5） μg/L, 5. 1 （2. 4-7. 8）μg/L and 2. 8（2. 2-5.7） μg/L. It also showed significant differences （U value were 246,419 and 790,P ＜0. 01）. The GP73 mRNA expression of PHC liver tissues（12. 64） was significant higher than normal liver tissues （1.00）. The critical values for GP73 and AFP was determined to be 123. 2 μg/L and 10. 6 μg/L through the 8OC curves. Under the critical value the sensitivity of GP73 and AFP were 65.8% and 53.4% ,and the specificity of CP73 and AFP were 95.3% and 92. 5% respectively. Joint detection could increase the sensitivity up to 79. 5%, and achieve the high specificity of 90. 7%. Conclusions As a new diagnositic marker of primary hepatic carcinoma, GP73 protein has the very good sensitivity and specificity. The GP73 mRNA in the whole blood sample could not be used for the diagnosis of PHC. But it woule be a good molecular marker for diagnosis of PHC in the liver tissue sample. The joint detection of GP73 and AFP could improve PHC diagnostic performance, and provide an effective approcach to the PHC high-risk screening.