miR-135a在胶质瘤中的表达变化及作用机制
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摘要
目的探讨miR-135a在胶质瘤中的表达及对胶质瘤细胞增殖、凋亡的影响,并探讨其相关分子机制。方法采用实时荧光定量PCR法检测60例的胶质瘤组织及相对应的癌旁组织、人胶质瘤细胞(U87、U251、A172)和人星形胶质细胞HA1800中miR-135a表达水平。将miR-135a mimics转染至U87细胞,采用CCK-8实验、流式细胞术观察U87细胞增殖活性及凋亡率的变化,采用Western blotting法观察U87细胞HOXA10蛋白的表达变化。结果胶质瘤组织较癌旁组织中miR-135a表达低(P<0.01);胶质瘤细胞U87、U251、A172中miR-135a表达较人星形胶质细胞HA1800低(P<0.01);随着胶质瘤病理分级的增高,miR-135a的表达逐渐降低(P均<0.01)。上调U87细胞miR-135a的表达后,细胞的增殖活性降低,凋亡率升高,HOXA10蛋白表达降低(P均<0.01)。结论 miR-135a在胶质瘤中呈低表达,miR-135a通过下调HOXA10基因在胶质瘤中发挥着抑癌作用。
Objective To investigate the expression of microRNA-135 a(miR-135 a) in the glioma and its effects on proliferation and apoptosis of glioma cells and its related molecular mechanism. Methods The quantitative real-time fluorescent PCR(qRT-PCR) was used to detect the expression of miR-135 a in 60 cases of glioma tissues and corresponding adjacent tissues of human glioma cells(U87, U251, and A172) and human astrocytes HA1800. After miR-135 a mimics were transfected into U87 cells, the proliferation activity and apoptotic rate of U87 cells were observed by CCK-8 assay and flow cytometry, and the expression of HOXA10 protein in U87 cells was observed by Western blotting. Results The expression of miR-135 a in the glioma tissues was lower than that in the adjacent tissues(P<0.01); the expression of miR-135 a in the glioma cells U87, U251 and A172 was lower than that in the human astrocytes HA1800(P<0.01); the expression of miR-135 a decreased gradually with the increase of pathological grade of glioma(P<0.01). Up-regulation of the expression of miR-135 a in U87 cells decreased the proliferation activity, increased the apoptotic rate, and decreased the expression of HOXA10 protein(all P<0.01).Conclusion The expression of miR-135 a is low in glioma, and miR-135 a plays an anti-cancer role in glioma by down-regulating HOXA10 gene.
Objective To investigate the expression of microRNA-135 a(miR-135 a) in the glioma and its effects on proliferation and apoptosis of glioma cells and its related molecular mechanism. Methods The quantitative real-time fluorescent PCR(qRT-PCR) was used to detect the expression of miR-135 a in 60 cases of glioma tissues and corresponding adjacent tissues of human glioma cells(U87, U251, and A172) and human astrocytes HA1800. After miR-135 a mimics were transfected into U87 cells, the proliferation activity and apoptotic rate of U87 cells were observed by CCK-8 assay and flow cytometry, and the expression of HOXA10 protein in U87 cells was observed by Western blotting. Results The expression of miR-135 a in the glioma tissues was lower than that in the adjacent tissues(P<0.01); the expression of miR-135 a in the glioma cells U87, U251 and A172 was lower than that in the human astrocytes HA1800(P<0.01); the expression of miR-135 a decreased gradually with the increase of pathological grade of glioma(P<0.01). Up-regulation of the expression of miR-135 a in U87 cells decreased the proliferation activity, increased the apoptotic rate, and decreased the expression of HOXA10 protein(all P<0.01).Conclusion The expression of miR-135 a is low in glioma, and miR-135 a plays an anti-cancer role in glioma by down-regulating HOXA10 gene.
引文
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[19] Chen Y, Zhang J, Wang H, et al. miRNA-135a promotes breast cancer cell migration and invasion by targeting HOXA10[J]. BMC Cancer, 2012,12:111.
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[23] Li B, McCrudden CM, Yuen HF, et al. CD133 in brain tumor: the prognostic factor[J]. Oncotarget, 2017,8(7):11144-11159.
[24] Kim JW, Kim JY, Kim JE, et al. HOXA10 is associated with temozolomide resistance through regulation of the homologous recombinant DNA repair pathway in glioblastoma cell lines[J]. Genes Cancer, 2014,5(5-6):165-174.
[2] Wu M, Fan Y, Lv S, et al. Incristine and temozolomide combined chemotherapy for the treatment of glioma: a comparison of solid lipid nanoparticles and nanostructured lipid carriers for dual drugs delivery[J]. Drug Deliv, 2016,23(8):2720-2725.
[3] Sorel O, DewalsBG. MicroRNAs in large herpesvirus DNA genomes: recent advances[J]. Biomol Concepts, 2016,7(4):229-239.
[4] Di Leva G, Garofalo M, Croce CM. MicroRNAs in cancer[J]. Annu Rev Pathol, 2014,9:287-314.
[5] Zhang Y, Dutta A, Abounader R. The role of microRNAs in glioma initiation and progression[J]. Front Biosci (Landmark Ed), 2012,17:700-712.
[6] Palumbo S, Miracco C, Pirtoli L, et al. Emerging roles of microRNA in modulating cell-death processes in malignant glioma[J]. J Cell Physiol, 2014,229(3):277-286.
[7] 荀静宇,李福军,孙晨,等.miR-135a在消化系统肿瘤中的研究进展[J].胃肠病学和肝病学杂志,2017,26(7):816-818.
[8] Hemmesi K, Squadrito ML, Mestdagh P, et al. miR-135a Inhibits Cancer Stem Cell-Driven Medulloblastoma Development by Directly Repressing Arhgef6 Expression[J]. Stem Cells, 2015,33(5):1377-1389.
[9] Ahmad A, Zhang W, Wu M, et al. Tumor-suppressive miRNA-135a inhibits breast cancer cell proliferation by targeting ELK1 and ELK3 oncogenes[J]. Genes Genomics, 2018,40(3):243-251.
[10] Wang LX, Kang ZP, Yang ZC, et al. MicroRNA-135a Inhibits Nasopharyngeal Carcinoma Cell Proliferation Through Targeting Interleukin-17[J]. Cell Physiol Biochem, 2018,46(6):2232-2238.
[11] Zhang C, Chen X, Chen X, et al. miR-135a acts as a tumor suppressor in gastric cancer in part by targeting KIFC1[J]. Onco Targets Ther, 2016,9:3555-3563.
[12] von Felden J, Heim D, Schulze K, et al. High expression of micro RNA-135A in hepatocellular carcinoma is associated with recurrence within 12 months after resection[J]. BMC Cancer, 2017,17(1):60.
[13] Zeng YB, Liang XH, Zhang GX3, et al. miRNA-135a promotes hepatocellular carcinoma cell migration and invasion by targeting forkhead box O1[J]. Cancer Cell Int, 2016,16:63.
[14] Mao XP, Zhang LS, Huang B, et al. Mir-135a enhances cellular proliferation through post-transcriptionally regulating PHLPP2 and FOXO1 in human bladder cancer[J]. J Transl Med, 2015,13:86.
[15] Zhou W, Li X, Liu F, et al. MiR-135a promotes growth and invasion of colorectal cancer via metastasis suppressor 1 in vi 4 vtro[J]. Acta Biochim Biophys Sin (Shanghai), 2012,44(10):838-846.
[16] 汤伟伟,万贵平,万一聪,等.miR-135a对卵巢上皮性癌细胞中HOXA10基因表达及细胞增殖和凋亡的影响[J].中华妇产科杂志,2013,48(5):364-369.
[17] Tang W, Jiang Y, Mu X, et al. MiR-135a functions as a tumor suppressor in epithelial ovarian cancer and regulates HOXA10 expression[J]. Cell Signal, 2014,26(7):1420-1426.
[18] Guo LM, Ding GF, Xu W, et al. MiR-135a-5p represses proliferation of HNSCC by targeting HOXA10[J]. Cancer Biol Ther, 2018,26:1-28.
[19] Chen Y, Zhang J, Wang H, et al. miRNA-135a promotes breast cancer cell migration and invasion by targeting HOXA10[J]. BMC Cancer, 2012,12:111.
[20] 王斌,张逊.同源盒基因与肿瘤的研究进展[J].医学综述,2017,23(7):1329-1333.
[21] Vastrad B, Vastrad C, Godavarthi A, et al. Molecular mechanisms underlying gliomas and glioblastoma pathogenesis revealed by bioinformatics analysis of microarray data[J]. Med Oncol, 2017,34(11):182.
[22] Duan R, Han L, Wang Q, et al. HOXA13 is a potential GBM diagnostic marker and promotes glioma invasion by activating the Wnt and TGF-β pathways[J]. Oncotarget, 2015,6(29):27778-27793.
[23] Li B, McCrudden CM, Yuen HF, et al. CD133 in brain tumor: the prognostic factor[J]. Oncotarget, 2017,8(7):11144-11159.
[24] Kim JW, Kim JY, Kim JE, et al. HOXA10 is associated with temozolomide resistance through regulation of the homologous recombinant DNA repair pathway in glioblastoma cell lines[J]. Genes Cancer, 2014,5(5-6):165-174.