miRNA表达与免疫细胞因子相关性在原发性肝癌早期诊断和预后中的作用
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摘要
目的:检测miR-146a、miR-224、miR-34c、miR-200a、miR-148b、miR-375六种miRNAs在原发性肝癌中的表达变化及其与HBs Ag、抗-HBs、HBe Ag、抗-HBe、抗-HBc和IL-12、IL-4、IL-6、IL-10、IFN-γ、TNF-α等炎症因子表达的相关性,以验证循环miRNA是否可作为理想的血源性新型生物标志物用于原发性肝癌的早期检测。方法:收集肝炎、肝硬化患者及健康对照组静脉血,并收集原发性肝癌患者癌组织和癌旁组织。提取总RNA后通过实时定量PCR检测并比较各组miRNA的相对表达水平,同时检测miRNA表达水平变化与血清肿瘤标志物AFP、CEA、CA19-9、CA125表达的关系;并检测miRNAs表达水平变化与HBs Ag、抗-HBs、HBe Ag、抗-HBe、抗-HBc和炎症因子IL-12、IL-4、IL-6、IL-10、IFN-γ、TNF-α表达相关性。结果:相对于健康组,miR-34c、miR-224、miR-146a在PHC组血清和组织中表达显著上调;miR-200a、miR-148b、miR-375在PHC组血清和组织中表达显著下调,差异具有统计学意义。HBs Ag与血清miR-375和miR-146a存在回归关系,miR-375随HBs Ag表达水平升高而降低,而miR-146a随HBs Ag表达升高而升高。IFN-γ与miR-146a存在回归关系,miR-146a随IFN-γ表达水平降低而升高,miR-375和miR-146a诊断能力大于CA19-9和AFP。结论:miR-146a、miR-224、miR-34c、miR-200a、miR-148b、miR-375在原发性肝癌血清和组织中存在表达差异,其中miR-375和miR-146a诊断能力优于AFP和CA19-9,血清miR-375和miR-146a可能成为新的肝癌早期诊断标志。
Objective: To detect the expression of six miRNAs for miR-146 a,miR-224,miR-34c,miR-200 a,miR-148 b,miR-375 level in primary hepatocellular carcinoma patients tissues and serum,and to explore the correlation between the expression of miRNA and inflammatory factors,in the hope of verification whether circulating miRNA could be used as an ideal blood borne biomarker for early detection of primary liver cancer. Methods: Venous blood samples were collected from patients with primary liver cancer,hepatitis,cirrhosis and healthy controls,cancer tissues and para cancerous tissues were collected from patients with cancer. After the total RNA was extracted,the relative expression levels of miRNA were detected by real-time quantitative PCR,and the relationship between the expression level of miRNA and the expression of serum tumor markers AFP,CEA,CA19-9 and CA125 was detected. The correlation between the expression level of miRNAs and the expression of HBs Ag,anti-HBs,HBe Ag,anti-HBe,anti-HBc and inflammatory factors IL-12,IL-4,IL-6,IL-10,IFN-γ,TNF-α was detected as well. Results: Compared with the healthy group,the expression of miR-34 c,miR-224 and miR-146 a in serum and tissues of PHC group was significantly up-regulated,and the expression of miR-200 a,miR-148 b and miR-375 in serum and tissues of PHC group was significantly down regulated. The difference was statistically significant. There was a regression relationship between HBs Ag and serum miR-375 and miR-146 a,and miR-375 decreased with the increase of HBs Ag expression,while miR-146 a increased with the increase of HBs Ag. There was a regression relationship between IFN-γ and miR-146 a,miR-146 a increased with the decrease of IFN-γ expression. The diagnostic ability of miR-375 and miR-146 a was greater than that of CA19-9 and AFP. Conclusion: MiR-146 a,miR-224,miR-34 c,miR-200 a,miR-148 b,miR-375 in serum and tissue of primary hepatocellular carcinoma are differentially express,among them,the diagnostic ability of miR-375 and miR-146 a is better than that of AFP and CA19-9,and serum miR-375 and miR-146 a may be the new markers for early diagnosis of PHC.
Objective: To detect the expression of six miRNAs for miR-146 a,miR-224,miR-34c,miR-200 a,miR-148 b,miR-375 level in primary hepatocellular carcinoma patients tissues and serum,and to explore the correlation between the expression of miRNA and inflammatory factors,in the hope of verification whether circulating miRNA could be used as an ideal blood borne biomarker for early detection of primary liver cancer. Methods: Venous blood samples were collected from patients with primary liver cancer,hepatitis,cirrhosis and healthy controls,cancer tissues and para cancerous tissues were collected from patients with cancer. After the total RNA was extracted,the relative expression levels of miRNA were detected by real-time quantitative PCR,and the relationship between the expression level of miRNA and the expression of serum tumor markers AFP,CEA,CA19-9 and CA125 was detected. The correlation between the expression level of miRNAs and the expression of HBs Ag,anti-HBs,HBe Ag,anti-HBe,anti-HBc and inflammatory factors IL-12,IL-4,IL-6,IL-10,IFN-γ,TNF-α was detected as well. Results: Compared with the healthy group,the expression of miR-34 c,miR-224 and miR-146 a in serum and tissues of PHC group was significantly up-regulated,and the expression of miR-200 a,miR-148 b and miR-375 in serum and tissues of PHC group was significantly down regulated. The difference was statistically significant. There was a regression relationship between HBs Ag and serum miR-375 and miR-146 a,and miR-375 decreased with the increase of HBs Ag expression,while miR-146 a increased with the increase of HBs Ag. There was a regression relationship between IFN-γ and miR-146 a,miR-146 a increased with the decrease of IFN-γ expression. The diagnostic ability of miR-375 and miR-146 a was greater than that of CA19-9 and AFP. Conclusion: MiR-146 a,miR-224,miR-34 c,miR-200 a,miR-148 b,miR-375 in serum and tissue of primary hepatocellular carcinoma are differentially express,among them,the diagnostic ability of miR-375 and miR-146 a is better than that of AFP and CA19-9,and serum miR-375 and miR-146 a may be the new markers for early diagnosis of PHC.
引文
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[9]Dhayat SA,Mardin WA,K9hler G,et al.The microRNA-200family--a potential diagnostic marker in hepatocellular carcinoma?[J].J Surg Oncol,2014,110(4):430-438.
[10]Ziari K,Zarea M,Gity M,et al.Downregulation of miR-148b as biomarker for early detection of hepatocellular carcinoma and may serve as a prognostic marker[J].Tumour Biol,2016,37(5):5765-5768.
[11]Mirzaei HR,Sahebkar A,Mohammadi M,et al.Circulating microRNAs in hepatocellular carcinoma:potential diagnostic and prognostic biomarkers[J].Curr Pharm Des,2016,22(34):5257-5269.
[2]陈彪,徐细明.miRNA在原发性肝癌中的研究进展[J].临床肿瘤学杂志,2012,17(10):946-950.Chen B,Xu XM.Progression of mi RNA in the research of primary liver cancer[J].Chin Clin Oncol,2012,17(10):946-950.
[3]米旭光,刘磊,李首庆,等.早期肝细胞癌患者循环microRNA-199a/b-3p的检测及临床意义[J].中国免疫学杂志,2015,31(5):683-685,689.Mi XG,Liu L,Li SQ,et al.Value of serum microRNA-199a/b-3p on early diagnosis of hepatocellular carcinoma[J].Chin J Immunol,2015,31(5):683-685,689.
[4]Marco D'Anzeo,Luca Faloppi,Mario Scartozzi,et al.The role of micro-RNAs in hepatocellular carcinoma:from molecular biology to treatment[J].Molecules,2014,19:6393-6406.
[5]李飞,李德新,周广朋.miR-634在肝癌中的表达及对肝癌细胞生物学行为的影响[J].中国免疫学杂志,2016,32(8):1160-1164.Li F,Li DX,Zhou GP.Expression of miR-634 in hepatocellular carcinoma and its effect on biological behavior of Hepatocellular carcinoma cells[J].Chin J Immunol,2016,32(8):1160-1164.
[6]Chamani F,Sadeghizadeh M,Masoumi M,et al.Evaluation of MiR-34 family and DNA methyltransferases 1,3A,3B gene expression levels in hepatocellular carcinoma following treatment with dendrosomal nanocurcumin.[J].Asian Pac J Cancer Prev,2016,17(S3):219-224.
[7]Shen SN,Wang LF,Jia YF,et al.Upregulation of microRNA-224 is associated with aggressive progression and poor prognosis in human cervical cancer[J].Diagn Pathol,2013,8:69.
[8]Lin CW,Chang YL,Chang YC,et al.microRNA-135b promotes lung cancer metastasis by regulating multiple targets in the Hippo pathway and LZTS1[J].Nat Commun,2013,4:1877.
[9]Dhayat SA,Mardin WA,K9hler G,et al.The microRNA-200family--a potential diagnostic marker in hepatocellular carcinoma?[J].J Surg Oncol,2014,110(4):430-438.
[10]Ziari K,Zarea M,Gity M,et al.Downregulation of miR-148b as biomarker for early detection of hepatocellular carcinoma and may serve as a prognostic marker[J].Tumour Biol,2016,37(5):5765-5768.
[11]Mirzaei HR,Sahebkar A,Mohammadi M,et al.Circulating microRNAs in hepatocellular carcinoma:potential diagnostic and prognostic biomarkers[J].Curr Pharm Des,2016,22(34):5257-5269.