非小细胞肺癌早期诊断的潜在生物标志物:血浆miRNA-23a和miRNA-451
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摘要
目的研究血浆中miRNA23-a及miRNA-451作为潜在生物标志物在非小细胞肺癌(NSCLC)中早期诊断的价值。方法选取本院50例NSCLC病例和50例健康对照,通过实时荧光定量PCR(qRT-PCR)测定组织和血浆中miRNA23-a及miRNA-451的表达水平,分析miRNA23-a及miRNA-451在组织和血浆中的表达差异及相关性。同时使用受试者工作特征曲线(ROC曲线)和线下面积(AUC)评估miRNA23-a、miRNA-451及CEA在NSCLC中的诊断能力。通过瞬时转染技术,在NSCLC细胞系A549中分别抑制miRNA-23a和miRNA-451的表达,使用qRT-PCR检测SPRY2 mRNA及MIF mRNA的表达水平。结果miRNA-23a及miRNA-451在NSCLC组织中的表达与吸烟状态、肿瘤大小、淋巴结转移和TNM分期显著相关(P<0.05)。与对照组相比,miRNA-23a在NSCLC患者的肿瘤组织和血浆中显著升高,而miRNA-451明显低于对照组。这2种miRNA的水平在肿瘤组织与血浆中表现出相同的表达趋势,呈高度相关。ROC分析显示,miRNA-23a和miRNA-451较之传统的肿瘤标志物CEA,AUCROC、灵敏度和特异度均显著高于后者(P<0.05)。当miRNA23-a及miRNA-451联合检测时,AUCROC/灵敏度/特异度均得到改善,分别为0.900/78%/86%。在NSCLC细胞系A549中分别抑制miRNA-23a和miRNA-451的表达后,SPRY2 mRNA及MIF mRNA表达水平显著升高。结论 miRNA-23a和miRNA-451可用作NSCLC早期诊断的候选标志物,这2种标志物联合检测对NSCLC诊断更有效。
Objective To study the value of plasma miRNA23-a and miRNA-451 as potential biomarkers for early diagnosis of non-small cell lung cancer(NSCLC). Methods Fifty patients with NSCLC and 50 healthy control subjects were recruited for testing the plasma levels of miRNA23-a and miRNA-451 and their expression levels in the tumor tissues using qRT-PCR. The correlations of the plasma levels of miRNA23-a and miRNA-451 with their expressions in the tumor tissues were analyzed.The diagnostic power of CEA, miRNA23-a and miRNA-451 for NSCLC was evaluated using the receiver-operating characteristics(ROC) curves and the area under the ROC curves(AUC). In the NSCLC cell line A549, we tested the effect of inhibition of miRNA-23 a and miRNA-451 on the expression levels of SPRY2 and MIF mRNA using qRT-PCR. Results The expression levels of miRNA-23 a and miRNA-451 in NSCLC tissues was significantly associated with smoking, tumor size,lymph node metastasis and TNM stage(P<0.05). Compared with those in the control group, miRNA-23 a level was significantly increased while miRNA-451 was significantly down-regulated in the tumor tissues and plasma of NSCLC patients. The plasma levels of miRNA-23 a and miRNA-45 were strongly correlated with their expression levels in the tumor tissues. ROC analysis showed that for the diagnosis of NSCLC, the AUC, sensitivity and specificity of either miRNA-23 a or miRNA-451 were significantly higher than those of CEA(P<0.05). The combination of miRNA23-a and miRNA-451 markedly improved the AUC(0.900), sensitivity(78%) and specificity(86%) for the diagnosis. In A549 cells, inhibition of miRNA23-a and miRNA-451 resulted in significantly increased expression levels of SPRY2 mRNA and MIF mRNA, respectively. Conclusions miRNA-23 a and miRNA-451 can be used as potential biomarkers for early diagnosis of NSCLC, and their combined detection can be more effective for the diagnosis.
Objective To study the value of plasma miRNA23-a and miRNA-451 as potential biomarkers for early diagnosis of non-small cell lung cancer(NSCLC). Methods Fifty patients with NSCLC and 50 healthy control subjects were recruited for testing the plasma levels of miRNA23-a and miRNA-451 and their expression levels in the tumor tissues using qRT-PCR. The correlations of the plasma levels of miRNA23-a and miRNA-451 with their expressions in the tumor tissues were analyzed.The diagnostic power of CEA, miRNA23-a and miRNA-451 for NSCLC was evaluated using the receiver-operating characteristics(ROC) curves and the area under the ROC curves(AUC). In the NSCLC cell line A549, we tested the effect of inhibition of miRNA-23 a and miRNA-451 on the expression levels of SPRY2 and MIF mRNA using qRT-PCR. Results The expression levels of miRNA-23 a and miRNA-451 in NSCLC tissues was significantly associated with smoking, tumor size,lymph node metastasis and TNM stage(P<0.05). Compared with those in the control group, miRNA-23 a level was significantly increased while miRNA-451 was significantly down-regulated in the tumor tissues and plasma of NSCLC patients. The plasma levels of miRNA-23 a and miRNA-45 were strongly correlated with their expression levels in the tumor tissues. ROC analysis showed that for the diagnosis of NSCLC, the AUC, sensitivity and specificity of either miRNA-23 a or miRNA-451 were significantly higher than those of CEA(P<0.05). The combination of miRNA23-a and miRNA-451 markedly improved the AUC(0.900), sensitivity(78%) and specificity(86%) for the diagnosis. In A549 cells, inhibition of miRNA23-a and miRNA-451 resulted in significantly increased expression levels of SPRY2 mRNA and MIF mRNA, respectively. Conclusions miRNA-23 a and miRNA-451 can be used as potential biomarkers for early diagnosis of NSCLC, and their combined detection can be more effective for the diagnosis.
引文
[1] White NM, Fatoohi E, Metias M, et al. Metastamirs:a stepping stone towards improved cancer management[J]. Nat Rev Clin Oncol, 2011,8:75-84.
[2] Metias SM, Lianidou E, Yousef GM. MicroRNAs in clinical oncology:at the crossroads between promises and problems[J]. J Clin Pathol, 2009, 62:771-6.
[3] Mitchell PS, Parkin RK, Kroh EM, et al. Circulating microRNAs as stable blood-based markers for cancer detection[J]. Proc Natl Acad Sci USA, 2008, 105:10513-8.
[4] Wu Q, Lu Z, Li H, et al. Next-generation sequencing of microRNAs for breast cancer detection[J]. J Biomed Biotechnol, 2011, 2011:597145.
[5] Wu Q, Wang C, Lu Z, et al. Analysis of serum genome-wide microRNAs for breast cancer detection[J]. Clin Chim Acta, 2012,413:1058-65.
[6] ZS Wu, Q Wu, CQ Wang, et al. MiR-339-5p inhibits breast cancer cell migration and invasion in vitro and may be a potential biomarker for breast cancer prognosis[J]. BMC Cancer, 2010, 10:542-51.
[7] Nakatani F, Ferracin M, Manara MC, et al. miR-34a predicts survival of Ewing's sarcoma patients and directly influences cell chemo-sensitivity and malignancy[J]. J Pathol, 2012, 226:796-805.
[8]Frampton AE, Castellano L, Colombo T, et al. Integrated molecular analysis to investigate the role of microRNAs in pancreatic tumour growth and progression[J]. Lancet, 2015, 385(Suppl. 1), S37.
[9] Komatsu S, Ichikawa D, Kawaguchi T, et al. Plasma microRNA profiles:Identification of miR-23a as a novel biomarker for chemoresistance in esophageal squamous cell carcinoma[J].Oncotarge, 2016, 7:62034-48.
[10]Pu M, Li C, Q i X, et al. MiR-1254 suppresses HO-1 expression through seed region-dependent silencing and non-seed interaction with TFAP2A transcript to attenuate NSCLC growth[J]. PLOS Genet, 2017, 13:e1006896.
[11]Qu WQ, Liu L, YuZ. Clinical value of microRNA-23a upregulation in non-small cell lung cancer[J]. Int J Clin Exp Med, 2015, 8(8):13598-603.
[12]Wang R, Wang ZX, Yang JS, et al. MicroRNA-451 functions as a tumor suppressor in human non-small cell lung cancer by targeting ras-related protein 14(RAB14)[J]. Oncogene, 2011, 30:2644-58.
[13]Chandramouli S, Yu CY, Yusoff P, et al.Tesk1 interacts with Spry2 to abrogate its inhibition of ERK phosphorylation downstream of receptor tyrosine kinase signaling[J]. J Biol Chem, 2008, 283:167991.
[14]Wu G, Qin XQ, Guo JJ, et al.AKT/ERK activation is associated with gastric cancer cell resistance to paclitaxel[J]. Int J Clin Exp Pathol,2014, 7:144958.
[15]Livak KJ, Schmittgen TD. Analysis of relative gene expression data using realtime quantitative PCR and the 2(Dalta Dalta C(T))method[J]. Methods, 2001, 25:4028.
[16]YingJia Li, Hui Chen, PengFei She, et al. microRNA23a promotes cell growth and metastasis in gastric cancer via targeting SPRY2mediated ERK signaling[J]. Oncol Letters, 2018, 15:8433-41.
[17]Richard V, Kindt N, Saussez S. Macrophagemigration inhibitory factor involvement inbreast cancer(Review)[J]. Int J Oncol, 2015,47:1627-33.
[18]Liu N, Jiang N, Guo R, et al. MiR-451 inhibits cell growth and invasion by targeting MIF and is associated with survival in nasopharyngeal carcinoma[J]. Mol Cancer, 2013, 12:123.
[19]Dore LC, Amigo JD, Dos SC, et al. A GATA-1-regulated microRNA locus essential for erythropoiesis[J]. Proc Natl Acad Sci, 2008, 105:3333-8.
[20]MD Jansson, ND Damas, M Lees, et al. miR-339-5p regulates the p53 tumor-suppressor pathway by targeting MDM2[J]. Oncogene,2015, 34:1908-18.
[21]YL Wang, CM Chen, XM Wang, et al. Effects of miR-339-5p on invasion and prognosis of hepatocellular carcinoma[J]. Clin Res Hepatol Gastroenterol, 2016, 40:51-6.
[22]杨艳. miRNA-21、miRNA-451在食管鳞癌患者血清中的表达和放疗疗效评估中的作用[D].安徽:蚌埠医学院, 2017:1-46.
[23]包佑根,巢海潮,朱遵伟,等. miRNA-451在膀胱癌组织中的表达及其临床意义[J].实用癌症杂志, 2016, 31(11):1754-8.
[24]Wang N, Tan HY, Feng YG, et al. microRNA-23a in human cancer:Its roles, mechanisms and therapeutic relevance[J]. Cancers, 2019,11(1):7.
[25]Li ZZ, Li YY, Fu J, et al. Clinical utility of microRNA-451 as diagnostic biomarker for human cancers[J]. Biosci Rep, 2019, 39(1):BSR20180653.
[26]Yong FL, Law CW, Wang CW. Potentiality of a triple microRNA classifier:MiR-193a-3p, miR-23a and miR-338-5p for early detection of colorectal cancer[J]. BMC Cancer, 2013, 13:280.
[27]Redova M, Poprach A, Nekvindova J, et al. Circulating miR-378 and miR-451 in serum are potential biomarkers for renal cell carcinoma[J]. J Transl Med, 2012, 10:55.
[28]Liu C, Liang S, Xiao S, et al. MicroRNA-27b inhibits Spry2expression and promotes cell invasion in glioma U251 cells[J].Oncol Letters, 2015, 9:1393-7.
[29]Chandramouli S, Yu CY, Yusoff P, et al.Tesk1 interacts with Spry2 to abrogate its inhibition of ERK phosphorylation downstream of receptor tyrosine kinase signaling[J]. J Biol Chem, 2008, 283:1679-91.
[30]李锦意,柳莉丹,陈永锋. Sprouty蛋白与疾病关系的研究进展[J].皮肤性病诊疗学志, 2018, 25(2):107-10.
[31]Kindt N, Preillon J, Kaltner H, et al:Macrophage migration inhibitory factor in head and neck squamous cell carcinoma:Clinical and experimental studies[J]. J Cancer Res Clin Oncol, 2013, 139:727-37.
[32]Wang D, Luo L, Chen W, et al. Significance of the vascular endothelial growth factor and the macrophage migration inhibitory factor in the progression of hepatocellular carcinoma[J]. Oncol Rep,2014, 31:11991204.
[33] Krockenberger M, Kranke P, Hausler S, et al. Macrophage migration-inhibitory factor levels in serum of patients with ovarian cancer correlates with poor prognosis[J]. Anticancer Res, 2012, 32:5233-8.
[34]赵娣. CD163与MIF在非小细胞肺癌的表达及对血管生成的影响[D].河南:郑州大学, 2018:1-49.
[2] Metias SM, Lianidou E, Yousef GM. MicroRNAs in clinical oncology:at the crossroads between promises and problems[J]. J Clin Pathol, 2009, 62:771-6.
[3] Mitchell PS, Parkin RK, Kroh EM, et al. Circulating microRNAs as stable blood-based markers for cancer detection[J]. Proc Natl Acad Sci USA, 2008, 105:10513-8.
[4] Wu Q, Lu Z, Li H, et al. Next-generation sequencing of microRNAs for breast cancer detection[J]. J Biomed Biotechnol, 2011, 2011:597145.
[5] Wu Q, Wang C, Lu Z, et al. Analysis of serum genome-wide microRNAs for breast cancer detection[J]. Clin Chim Acta, 2012,413:1058-65.
[6] ZS Wu, Q Wu, CQ Wang, et al. MiR-339-5p inhibits breast cancer cell migration and invasion in vitro and may be a potential biomarker for breast cancer prognosis[J]. BMC Cancer, 2010, 10:542-51.
[7] Nakatani F, Ferracin M, Manara MC, et al. miR-34a predicts survival of Ewing's sarcoma patients and directly influences cell chemo-sensitivity and malignancy[J]. J Pathol, 2012, 226:796-805.
[8]Frampton AE, Castellano L, Colombo T, et al. Integrated molecular analysis to investigate the role of microRNAs in pancreatic tumour growth and progression[J]. Lancet, 2015, 385(Suppl. 1), S37.
[9] Komatsu S, Ichikawa D, Kawaguchi T, et al. Plasma microRNA profiles:Identification of miR-23a as a novel biomarker for chemoresistance in esophageal squamous cell carcinoma[J].Oncotarge, 2016, 7:62034-48.
[10]Pu M, Li C, Q i X, et al. MiR-1254 suppresses HO-1 expression through seed region-dependent silencing and non-seed interaction with TFAP2A transcript to attenuate NSCLC growth[J]. PLOS Genet, 2017, 13:e1006896.
[11]Qu WQ, Liu L, YuZ. Clinical value of microRNA-23a upregulation in non-small cell lung cancer[J]. Int J Clin Exp Med, 2015, 8(8):13598-603.
[12]Wang R, Wang ZX, Yang JS, et al. MicroRNA-451 functions as a tumor suppressor in human non-small cell lung cancer by targeting ras-related protein 14(RAB14)[J]. Oncogene, 2011, 30:2644-58.
[13]Chandramouli S, Yu CY, Yusoff P, et al.Tesk1 interacts with Spry2 to abrogate its inhibition of ERK phosphorylation downstream of receptor tyrosine kinase signaling[J]. J Biol Chem, 2008, 283:167991.
[14]Wu G, Qin XQ, Guo JJ, et al.AKT/ERK activation is associated with gastric cancer cell resistance to paclitaxel[J]. Int J Clin Exp Pathol,2014, 7:144958.
[15]Livak KJ, Schmittgen TD. Analysis of relative gene expression data using realtime quantitative PCR and the 2(Dalta Dalta C(T))method[J]. Methods, 2001, 25:4028.
[16]YingJia Li, Hui Chen, PengFei She, et al. microRNA23a promotes cell growth and metastasis in gastric cancer via targeting SPRY2mediated ERK signaling[J]. Oncol Letters, 2018, 15:8433-41.
[17]Richard V, Kindt N, Saussez S. Macrophagemigration inhibitory factor involvement inbreast cancer(Review)[J]. Int J Oncol, 2015,47:1627-33.
[18]Liu N, Jiang N, Guo R, et al. MiR-451 inhibits cell growth and invasion by targeting MIF and is associated with survival in nasopharyngeal carcinoma[J]. Mol Cancer, 2013, 12:123.
[19]Dore LC, Amigo JD, Dos SC, et al. A GATA-1-regulated microRNA locus essential for erythropoiesis[J]. Proc Natl Acad Sci, 2008, 105:3333-8.
[20]MD Jansson, ND Damas, M Lees, et al. miR-339-5p regulates the p53 tumor-suppressor pathway by targeting MDM2[J]. Oncogene,2015, 34:1908-18.
[21]YL Wang, CM Chen, XM Wang, et al. Effects of miR-339-5p on invasion and prognosis of hepatocellular carcinoma[J]. Clin Res Hepatol Gastroenterol, 2016, 40:51-6.
[22]杨艳. miRNA-21、miRNA-451在食管鳞癌患者血清中的表达和放疗疗效评估中的作用[D].安徽:蚌埠医学院, 2017:1-46.
[23]包佑根,巢海潮,朱遵伟,等. miRNA-451在膀胱癌组织中的表达及其临床意义[J].实用癌症杂志, 2016, 31(11):1754-8.
[24]Wang N, Tan HY, Feng YG, et al. microRNA-23a in human cancer:Its roles, mechanisms and therapeutic relevance[J]. Cancers, 2019,11(1):7.
[25]Li ZZ, Li YY, Fu J, et al. Clinical utility of microRNA-451 as diagnostic biomarker for human cancers[J]. Biosci Rep, 2019, 39(1):BSR20180653.
[26]Yong FL, Law CW, Wang CW. Potentiality of a triple microRNA classifier:MiR-193a-3p, miR-23a and miR-338-5p for early detection of colorectal cancer[J]. BMC Cancer, 2013, 13:280.
[27]Redova M, Poprach A, Nekvindova J, et al. Circulating miR-378 and miR-451 in serum are potential biomarkers for renal cell carcinoma[J]. J Transl Med, 2012, 10:55.
[28]Liu C, Liang S, Xiao S, et al. MicroRNA-27b inhibits Spry2expression and promotes cell invasion in glioma U251 cells[J].Oncol Letters, 2015, 9:1393-7.
[29]Chandramouli S, Yu CY, Yusoff P, et al.Tesk1 interacts with Spry2 to abrogate its inhibition of ERK phosphorylation downstream of receptor tyrosine kinase signaling[J]. J Biol Chem, 2008, 283:1679-91.
[30]李锦意,柳莉丹,陈永锋. Sprouty蛋白与疾病关系的研究进展[J].皮肤性病诊疗学志, 2018, 25(2):107-10.
[31]Kindt N, Preillon J, Kaltner H, et al:Macrophage migration inhibitory factor in head and neck squamous cell carcinoma:Clinical and experimental studies[J]. J Cancer Res Clin Oncol, 2013, 139:727-37.
[32]Wang D, Luo L, Chen W, et al. Significance of the vascular endothelial growth factor and the macrophage migration inhibitory factor in the progression of hepatocellular carcinoma[J]. Oncol Rep,2014, 31:11991204.
[33] Krockenberger M, Kranke P, Hausler S, et al. Macrophage migration-inhibitory factor levels in serum of patients with ovarian cancer correlates with poor prognosis[J]. Anticancer Res, 2012, 32:5233-8.
[34]赵娣. CD163与MIF在非小细胞肺癌的表达及对血管生成的影响[D].河南:郑州大学, 2018:1-49.
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