Sha M, Wang B, Xiao L, Ye J, Wang J, Luan Z Y
Department of Clinical Medicine, Taizhou People's Hospital Affiliated of Nantong University of Medicine, Tai Zhou, 225300, China.
Department of Infectious Disease, Taizhou People's Hospital Affiliated of Nantong University of Medicine, Tai Zhou 225300, China.
Zhonghua Gan Zang Bing Za Zhi. 2017 Dec 20;25(12):920-926. doi: 10.3760/cma.j.issn.1007-3418.2017.12.007.
To investigate the expression of miR-212 and miR-132 in the serum of patients with primary liver cancer and their targeted regulation of GP73. The patients with liver cancer, chronic hepatitis B, or liver cirrhosis who were hospitalized in Taizhou People's Hospital from January 2015 to December 2016 were enrolled, and healthy volunteers were also enrolled as controls. Quantitative real-time PCR was used to measure the serum levels of miR-212 and miR-132, and the association between the expression of serum miR-212 and miR-132 and the clinicopathological features of patients with liver cancer was analyzed. A Spearman's rank correlation analysis was used to analyze the correlation between serum miR-212/miR-132 and GP73. Western blot was used to measure the protein expression of GP73, and MTT assay was used to measure the survival rate of cells. The Levene's homogeneity of variance test was used for data analysis. The independent samples t-test was used for comparison of means between two samples, and ANOVA was used for comparison of means between multiple samples. A total of 90 patients with liver cancer, 60 with chronic hepatitis B, 68 with liver cirrhosis, and 100 healthy volunteers were enrolled. The relative expression levels of miR-212 and miR-132 in serum were 0.046 6 ± 0.024 7 and 0.005 9 ± 0.003 0 in the patients with liver cancer, 0.979 7 ± 0.259 5 and 1.001 8 ± 0.249 9 in the healthy volunteers, 0.588 2 ± 0.216 5 and 0.345 7 ± 0.233 8 in the patients with hepatitis, and 0.313 8 ± 0.153 3 and 0.080 1 ± 0.042 66 in the patients with liver cirrhosis. Compared with the normal controls, all patients had significant reductions in the expression of serum miR-212 ( = 10.26, 20.86, and 35.80, all < 0.01) and miR-132 ( = 16.55, 36.09, and 39.85, all < 0.01). In the patients with liver cancer, the relative expression of miR-212 and miR-132 was negatively correlated with alpha-fetoprotein (miR-212: = -4.46, < 0.01; miR-132: = -4.83, < 0.01), TNM stage (miR-212: = 6.569, < 0.01; miR-132: = 7.31, < 0.01), degree of tumor differentiation (miR-212: = 5.268, < 0.01; miR-132: = 5.914, < 0.01), and presence of portal vein tumor thrombus (miR-212: = 5.16, < 0.01; miR-132: = 3.681, < 0.01), while it was not correlated with tumor size (miR-212: = 0.687, > 0.05; miR-132: = 0.887, > 0.05). In addition, serum miR-212 and miR-132 were negatively correlated with GP73 in the patients with liver cancer (miR-212: (s) = -0.709, < 0.01; miR-132: (s) = -0.877, < 0.01). Overexpression of miR-212 or miR-132 in HepG2 cells significantly inhibited the activity and expression of 3'-UTR, and interference of miR-212 or miR-132 significantly increased the activity and expression of 3'-UTR in GP73. Overexpression of GP73 reversed the reduction in survival rate of hepatoma cells induced by the overexpression of miR-212 or miR-132. Patients with liver cancer have a significant reduction in the expression of miR-212 and miR-132 in serum, which is closely associated with the development, progression, and metastasis of liver cancer, and miR-212 and miR-132 in hepatoma cells inhibit the growth of liver cancer by targeted regulation of GP73 expression.
探讨原发性肝癌患者血清中miR-212和miR-132的表达及其对GP73的靶向调控作用。选取2015年1月至2016年12月在泰州市人民医院住院的肝癌、慢性乙型肝炎或肝硬化患者,并纳入健康志愿者作为对照。采用定量实时荧光定量PCR检测血清中miR-212和miR-132水平,分析血清miR-212和miR-132表达与肝癌患者临床病理特征的关系。采用Spearman等级相关分析血清miR-212/miR-132与GP73的相关性。采用蛋白质免疫印迹法检测GP73蛋白表达,MTT法检测细胞存活率。采用Levene方差齐性检验进行数据分析。两组样本均数比较采用独立样本t检验,多组样本均数比较采用方差分析。共纳入90例肝癌患者、60例慢性乙型肝炎患者、68例肝硬化患者和100例健康志愿者。肝癌患者血清中miR-212和miR-132的相对表达水平分别为0.046 6±0.024 7和0.005 9±0.003 0,健康志愿者分别为0.979 7±0.259 5和1.001 8±0.249 9,肝炎患者分别为0.588 2±0.216 5和0.345 7±0.233 8,肝硬化患者分别为0.313 8±0.153 3和0.080 1±0.042 66。与正常对照组相比,所有患者血清miR-212(P=10.26、20.86和35.80,均P<0.01)和miR-132(P=16.55、36.09和39.85,均P<0.01)表达均显著降低。肝癌患者中,miR-212和miR-132的相对表达与甲胎蛋白(miR-212:P=-4.46,P<0.01;miR-132:P=-4.83,P<0.01)、TNM分期(miR-212:P=6.569,P<0.01;miR-132:P=7.31,P<0.01)、肿瘤分化程度(miR-212:P=5.268,P<0.01;miR-132:P=5.914,P<0.01)及门静脉癌栓形成(miR-212:P=5.16,P<0.01;miR-132:P=3.681,P<0.01)呈负相关,而与肿瘤大小无关(miR-212:P=0.687,P>0.05;miR-