食管鳞状细胞癌组织中miRNA-29a、miRNA-29b、miRNA-29c、VEGF-A表达变化及其相关性
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摘要
目的观察食管鳞状细胞癌(ESCC)癌组织及癌旁组织中miRNA-29家族(miRNA-29a、miRNA-29b、miRNA-29c)及血管内皮生长因子A(VEGF-A)的相对表达量变化,并探讨他们之间的关系。方法收集手术切除并经病理检查确诊的ESCC癌组织及癌旁组织,应用qRT-PCR及免疫组织化学染色技术分别检测miRNA-29a、miRNA-29b、miRNA-29c、VEGF-A的相对表达量,并分析其相关关系;进一步应用生物信息学方法和双荧光素酶报告基因系统推测和验证miRNA-29a、miRNA-29b、miRNA-29c共同调控同一靶基因VEGF-A的分子调节机制。结果与癌旁组织比较,ESCC癌组织中miRNA-29a、miRNA-29b、miRNA-29c呈低表达(P均<0.05);VEGF-A呈高表达(P<0.05),且其之间呈负相关关系(miRNA-29a:r=-0.955;miRNA-29b:r=-0.950;miRNA-29c:r=-0.893,P均<0.01)。生物信息学分析结果发现,VEGF-A能够与miRNA-29a、miRNA-29b、miRNA-29c进行特异性结合,可能为miRNA-29a、miRNA-29b、miRNA-29c共同的靶基因。双荧光素酶报告基因系统验证了VEGF-A是miRNA-29a、miRNA-29b、miRNA-29c共同的靶基因。结论 ESCC癌组织中miRNA-29a、miRNA-29b、miRNA-29c低表达,VEGFA高表达;且他们之间具有负相关关系。VEGF-A为miRNA-29a、miRNA-29b、miRNA-29c共同的靶向调节基因。
Objective To observe the expression changes of miRNA-29 family( miRNA-29 a,miRNA-29 b,and miRNA-29 c) and vascular endothelial growth factor-A( VEGF-A) in the esophageal squamous cell carcinoma( ESCC) tissues and to investigate the relationships between them. Methods The expression levels of miRNA-29 a/b/c and VEGF-A were detected by qRT-PCR and immunohistochemistry( IHC),respectively,in the ESCC tissues and para-carcinoma tissues.The relationship between the expression of miRNA-29 a/b/c and VEGF-A was further analyzed. We used the bioinformatics method and dual luciferase reporter gene system experiment to predict and verify that miRNA-29 a/b/c could co-regulate the same target gene VEGF-A. Results The higher expression of VEGF-A and the lower expression of miRNA-29 a/b/c were detected in the ESCC tissues as compared with that of the para-carcinoma tissues. There was a significant negative correlation between the expression levels of miRNA-29 a/b/c and VEGF-A in ESCC tissues( miRNA-29 a: r =-0. 955,miRNA-29 b: r =-0. 950,miRNA-29 c: r =-0. 893; all P < 0. 01). Bioinformatics analysis showed that VEGF-A could specifically bind with miRNA-29 a/b/c,which could be the common target gene of miRNA-29 a/b/c. The dual luciferase reporter system validated that VEGF-A was a common target gene of miRNA-29 a/b/c. Conclusions The miRNA-29 a/b/c is low expressed,and VEGF-A is highly expressed in the ESCC tissues,and they are negatively correlated with each other.VEGF-A is a common target regulator gene of miRNA-29 a/b/c.
Objective To observe the expression changes of miRNA-29 family( miRNA-29 a,miRNA-29 b,and miRNA-29 c) and vascular endothelial growth factor-A( VEGF-A) in the esophageal squamous cell carcinoma( ESCC) tissues and to investigate the relationships between them. Methods The expression levels of miRNA-29 a/b/c and VEGF-A were detected by qRT-PCR and immunohistochemistry( IHC),respectively,in the ESCC tissues and para-carcinoma tissues.The relationship between the expression of miRNA-29 a/b/c and VEGF-A was further analyzed. We used the bioinformatics method and dual luciferase reporter gene system experiment to predict and verify that miRNA-29 a/b/c could co-regulate the same target gene VEGF-A. Results The higher expression of VEGF-A and the lower expression of miRNA-29 a/b/c were detected in the ESCC tissues as compared with that of the para-carcinoma tissues. There was a significant negative correlation between the expression levels of miRNA-29 a/b/c and VEGF-A in ESCC tissues( miRNA-29 a: r =-0. 955,miRNA-29 b: r =-0. 950,miRNA-29 c: r =-0. 893; all P < 0. 01). Bioinformatics analysis showed that VEGF-A could specifically bind with miRNA-29 a/b/c,which could be the common target gene of miRNA-29 a/b/c. The dual luciferase reporter system validated that VEGF-A was a common target gene of miRNA-29 a/b/c. Conclusions The miRNA-29 a/b/c is low expressed,and VEGF-A is highly expressed in the ESCC tissues,and they are negatively correlated with each other.VEGF-A is a common target regulator gene of miRNA-29 a/b/c.
引文
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[2]Parkin DM,Bray F,Ferlay J,et al.Global cancer statistics,2002[J].CA Cancer J Clin,2005,55(2):74-108.
[3]Jiang H,Zhang G,Wu JH,et al.Diverse roles of miR-29 in cancer(review)[J].Oncol Rep,2014,31(4):1509-1516.
[4]Cortez MA,Nicoloso MS,Shimizu M,et al.miR-29b and miR-125a regulate podoplanin and suppress invasion in glioblastoma[J].Gene Chromosome Canc,2010,49(11):981-990.
[5]Ru P,Steele R,Newhall P,et al.miRNA-29b suppresses prostate cancer metastasis by regulating epithelial-mesenchymal transition signaling[J].Mol Cancer Ther,2012,11(5):1166-1173.
[6]Heinzelmann J,Henning B,Sanjmyatav J,et al.Specific miRNA signatures are associated with metastasis and poor prognosis in clear cell renal cell carcinoma[J].World J Urol,2011,29(3):367-373.
[7]Zhao JJ,Lin J,Lwin T,et al.microRNA expression profile and identification of miR-29 as a prognostic marker and pathogenetic factor by targeting CDK6 in mantle cell lymphoma[J].Blood,2010,115(13):2630-2639.
[8]Zeng X,Xiang J,Wu M,et al.Circulating miR-17,miR-20a,miR-29c,and miR-223 combined as non-invasive biomarkers in nasopharyngeal carcinoma[J].PLo S One,2012,7(10):e46367.
[9]Pass HI,Goparaju C,Ivanov S,et al.Hsa-miR-29c*is linked to the prognosis of malignant pleural mesothelioma[J].Cancer Res,2010,70(5):1916-1924.
[10]Fang C,Zhu DX,Dong HJ,et al.Serum microRNAs are promising novel biomarkers for diffuse large B cell lymphoma[J].Ann Hematol,2012,91(4):553-559.
[11]Costache MI,Ioana M,Iordache S,et al.VEGF expression in pancreatic cancer and other malignancies:a review of the literature[J].Rom J Intern Med,2015,53(3):199-208.
[12]Frezzetti D,Gallo M,Roma C,et al.Vascular endothelial growth factor a regulates the secretion of different angiogenic factors in lung cancer cells[J].J Cell Physiol,2016,231(7):1514-1521.
[13]Yang SM,Huang CY,Shiue HS,et al.Joint effect of urinary total arsenic level and vegf-a genetic polymorphisms on the recurrence of renal cell carcinoma[J].PLo S One,2015,10(12):e145410.
[14]Wei W,Wang Y,Yu X,et al.Expression of TP53,BCL-2,and VEGFA genes in esophagus carcinoma and its biological significance[J].Med Sci Monit,2015,21(1):3016-3022.