摘要
目的:探讨血清miR-378a-5p表达对早期肝癌诊断价值及其对Huh-7细胞增殖和侵袭影响。方法:应用Real-time PCR检测64例早期肝癌患者和64例常规体检健康志愿者血清中miR-378a-5p表达量,应用受试者工作特征曲线(Receiver operating characteristic curve,ROC)分析血清miR-17-5p和AFP表达对早期肝癌的筛查效能,t检验分析其与患者临床病理参数的关系。采用pcDNA3.1构建过表达miR-378a-5p质粒(pcDNA-miR-378a-5p),实现Huh-7细胞miR-378a-5p过表达后,运用CCK-8和Transwell实验观察Huh-7细胞增殖和侵袭能力变化。结果:早期肝癌患者血清中miR-378a-5p表达量高于健康人(2.263±0.964 vs 0.936±0.645,t=9.155,P<0.001)。肿瘤较大(>5cm)、肝癌细胞分级较差(Ⅲ级和Ⅳ级)、TNM分期Ⅰb期的早期肝癌患者血清miR-378a-5p表达水平升高(P均<0.05)。ROC分析结果显示血清miR-378a-5p表达预测早期肝癌的曲线下面积为0.868(95%CI:0.806-0.931,P<0.001),当cut-off值为1.685,其敏感性和特异性为71.9%和91.6%;血清AFP预测早期肝癌的ROC曲线下面积为0.851(95%CI:0.776-0.925,P<0.001),当cut-off值为17.35,其敏感性和特异性为75%和100%;血清miR-378a-5p联合AFP检测对早期肝癌预测的曲线下面积为0.950(95%CI:0.914-0.986,P<0.001),当cut-off值为61.52,其敏感性和特异性为81.3%和100%,Z检验显示两者联合检测筛选早期肝癌的效能优于血清miR-378a-5p(Z=2.233,P=0.026)和AFP(Z=2.354,P=0.019)独立检测。CCK-8和Transwell实验结果表明Huh-7细胞miR-378a-5p过表达后其增殖和侵袭能力增强(P均<0.05)。结论:miR-378a-5p在早期肝癌患者血清中显著升高,并促进了肝癌细胞生长和侵袭,与AFP联合检测可有效提高早期肝癌的筛查效果。
Objective:To explore the value of serum miR-378a-5p expression in the diagnosis of early liver cancer and its influence on the proliferation and invasion of Huh-7 cells.Methods:Real-time PCR was used to detect the expression of miR-378a-5p in the serum of 64 patients with early liver cancer and 64 healthy volunteers during routine physical examination.ROC was used to analyze the diagnostic value of serum miR-17-5p and AFP expression for early liver cancer,and the relationship between the expression of miR-17-5p and the clinical pathological parameters of the patients was analyzed by t test.Use pcDNA3.1 to construct an overexpressing miR-378a-5p plasmid(pcDNA-miR-378a-5p)to achieve miR-378a-5p overexpression,then CCK-8 and Transwell experiments was used to observe the proliferation and invasiveness of Huh-7 cells.Results:The expression of miR-378a-5p in the serum of patients with early liver cancer was higher than that of healthy people(2.263±0.964 vs 0.936±0.645,t=9.155,P<0.001).The serum miR-378a-5p expression level of early liver cancer patients with larger tumors(>5 cm)/worse grades of liver cancer cells(gradeⅢandⅣ)/TNMⅠb stage was elevated(allP<0.05).ROC analysis showed the area under the curve of serum miR-378a-5p expression predicting early liver cancer was 0.868(95%CI:0.806-0.931,P<0.001),the cut-off value was 1.685,the sensitivity and specificity were 71.9%and 91.6%.The area under the ROC curve of serum AFP predicting early liver cancer is 0.851(95%CI:0.776-0.925,P<0.001),the cut-off value is 17.35,the sensitivity and specificity are 75%and 100%.The area under the curve of serum miR-378a-5p combined with AFP predicting early liver cancer is 0.950(95%CI:0.914-0.986,P<0.001),the cut-off value is 61.52,the sensitivity and specificity are 81.3%and 100%.The Z test showed the combined detection of miR-378a-5p and AFP is more effective than the independent detection of serum miR-378a-5p(Z=2.233,P=0.026)and AFP(Z=2.354,P=0.019)in screening early liver cancer.The results of CCK-8 and Transwell experiments showed that the proliferation and invasion ability of Huh-7 cells was enhanced after miR-378a-5p overexpression(allP<0.05).Conclusion:MiR-378a-5p is significantly elevated in the serum of patients with early liver cancer,and promotes the growth and invasion of liver cancer cells,combined detection with AFP can effectively improve the screening effect of early liver cancer.
作者
何勇勇
方长英
张青松
HE Yongyong;FANG Changying;ZHANG Qingsong(Department of internal medicine,Xuancheng Vocational and Technical College,Xuancheng 242000,China;Department of Clinical Laboratory,Xuancheng Central Hospital)
出处
《包头医学院学报》
CAS
2021年第7期17-22,共6页
Journal of Baotou Medical College
