目的 研究miR-30a-5p和靶基因NUAK1在非小细胞肺癌中的表达及对A549细胞增殖、迁移侵袭的影响。方法 收集2016-09至2019-03在保定市第一医院行手术切除治疗的非小细胞肺癌患者癌组织和癌旁组织标本64例,QPCR和Western blot分别检测非小细胞肺癌组织样本和细胞中miR-30a-5p和NUAK1的表达;TargetScan网站预测miR-30a-5p与NUAK1间的靶向关系,双荧光素酶报告基因实验和Western blot进行验证;MTT和Transwell实验检测miR-30a-5p及联合过表达NUAK1对A549细胞增殖、迁移和侵袭的影响。结果 非小细胞肺癌组织中miR-30a-5p的相对表达量(0.33±0.14)低于癌旁组织(1.00±0.21),差异有统计学意义(t=21.237,P＜0.05);非小细胞肺癌组织中NUAK1 mRNA表达(4.13±1.87)和蛋白表达(3.41±1.62)均高于癌旁组织(1.00±0.17、1.00±0.16),差异有统计学意义(t=13.335、11.844,P＜0.05)。非小细胞肺癌细胞系H460、H1299、A549中miR-30a-5p表达量(0.35±0.03、0.51±0.05、0.28±0.03)低于人正常肺上皮细胞BEAS-2B(1.00±0.11),差异有统计学意义(F=77.100,P＜0.05);H460、H1299、A549细胞中NUAK1 mRNA相对表达量(2.98±0.30、2.39±0.24、3.42±0.36)和蛋白相对表达量(3.06±0.31、2.27±0.23、4.12±0.41)均显著高于BEAS-2B细胞(1.00±0.09、1.00±0.11),差异有统计学意义(F=46.660、63.070,P＜0.05)。结论 miR-30a-5p可通过靶向NUAK1抑制非小细胞肺癌细胞A549增殖、迁移和侵袭,提示miR-30a-5p和NUAK1可能成为一种新的临床诊断和治疗非小细胞肺癌的靶点。
Abstract
Objective To investigate the expressions of miR-30a-5p and its target gene NUAK1 in non-small cell lung cancer, and the effects on the proliferation, migration and invasion of A549 cells.Methods The specimens of 64 cases of NSCLC were collected.QPCR and Western blot were used to detect the expressions of miR-30a-5p and NUAK1 in NSCLC tissues and cell lines. The relationship between miR-30a-5p and NUAK1 was predicted via TargetScan website and was verified by dual luciferase reporter assay and Western blot. The effects of miR-30a-5p on proliferation, migration and invasion of A549 were determined by MTT and Transwell assays.Results The expression of miR-30a-5p in NSCLC tissues (0.33±0.14) was significantly lower than in adjacent tissues (1.00±0.21) (t=21.237, P<0.05), while the expressions of NUAK1 mRNA (4.13±1.87) and protein (3.41±1.62) in NSCLC tissues were significantly higher than in adjacent tissues (1.00±0.17, 1.00±0.16) (t=13.335, 11.844, P<0.05). The expression of miR-30a-5p in NSCLC cells H460, H1299 and A549 (0.35±0.03, 0.51±0.05, 0.28±0.03) was significantly lower than in normal human lung epithelial cells BEAS-2B (1.00±0.08) (F=77.100, P<0.05). The expressions of NUAK1 mRNA (2.98±0.30, 2.39±0.24, 3.42±0.35) and protein (3.06±0.31, 2.27±0.23, 4.12±0.41) in H460, H1299 and A549 cells were significantly higher than in BEAS-2B cells(1.00±0.09, 1.00±0.11) (F=46.660, 63.070, P<0.05).Conclusions miR-30a-5p can inhibit the proliferation, migration and invasion of A549 cells by targeting NUAK1, suggesting that miR-30a-5p and NUAK1 may become new targets for diagnosis and treatment of NSCLC.
关键词
miR-30a-5p /
新式激酶家族1 /
非小细胞肺癌 /
增殖 /
迁移 /
侵袭
Key words
miR-30a-5p /
NUAK1 /
non-small cell lung cancer /
proliferation /
migration /
invasion
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参考文献
[1] Chen W, Zheng R, Baade P D, et al. Cancer statistics in China, 2015[J].CA Cancer J Clin,2016,66(2):115-32.
[2] Jemal A, Bray F, Center M M, et al. Global cancer statistics[J]. CA Cancer J Clin,2011,61(2):69-90.
[3] Zhu J J, Zeng Y Y, Li W, et al. CD73/NT5E is a target of miR-30a-5p and plays an important role in the pathogenesis of non-small cell lung cancer[J]. Mol Cancer,2017,16(1):34.
[4] 邵 彬,李惠平,朱 军, 等. 复发转移性乳腺癌患者血浆miRNA表达水平与临床病理特征、化疗疗效及预后关系的研究[J].癌症进展, 2015, 13(2):106-113.
[5] Zhu J J, Zeng Y Y, Xu Ch, et al. Expression profile analysis of microRNAs and downregulated miR-486-5p and miR-30a-5p in non-small cell lung cancer[J]. Oncol Rep,2015,34(4):1779-1786.
[6] Cai J H, Fang L Sh, Huang Y B, et al. miR-205 targets PTEN and PHLPP2 to augment AKT signaling and drive malignant phenotypes in non-small cell lung cancer[J]. Cancer Res,2013,73(17):5402-5415.
[7] Wang J, Tian X, Han R, et al. Downregulation of miR-486-5p contributes to tumor progression and metastasis by targeting protumorigenic ARHGAP5 in lung cancer[J]. Oncogene, 2014,33(9):1181-1189.
[8] Chen P,Li K, Liang Y, et al. High NUAK1 expression correlates with poor prognosis and involved in NSCLC cells migration and invasion[J]. Exp Lung Res,2013,39(1):9-17.
[9] Hou X, Liu J E, Liu W, et al. A new role of NUAK1: directly phosphorylating p53 and regulating cell proliferation[J]. Oncogene,2011,30(26):2933-2942.
[10] Leva G D, Briskin D, Croce C M. MicroRNA in cancer: New hopes for antineoplastic chemotherapy[J]. Ups J Med Sci,2012,117(2):202-216.
[11] Qian. miRNAs in Cancer Prevention and Treatment and as Molecular Targets for Natural Product Anticancer Agents[J]. Curr Cancer Drug Targets,2013,13(5):519-541.
[12] Tang R, Liang L, Luo D, et al. Downregulation of MiR-30a is Associated with Poor Prognosis in Lung Cancer[J]. Med Sci Monit,2015,21:2514-2520.
[13] Fu J,Xu X,Kang L, et al. miR-30a suppresses breast cancer cell proliferation and migration by targeting Eya2[J]. Biochem Biophys Res Commun,2014,445(2):314-319.
[14] Dai H, Kang B, Zuo D Y, et al. Effect of miR-30a-5p on the proliferation, apoptosis, invasion and migration of SMCC-7721 human hepatocellular carcinoma cells[J]. Zhonghua Gan Zang Bing Za Zhi,2014,22(12):915-920.
[15] Zhang Q, Tang Q, Qin D, et al. Role of microRNA 30a targeting insulin receptor substrate 2 in colorectal tumorigenesis[J]. Mol Cell Biol,2015,35(6):988-1000.
[16] Wang H Y, Li Y Y, Fu S, et al. MicroRNA-30a promotes invasiveness and metastasis in vitro and in vivo through epithelial-mesenchymal transition and results in poor survival of nasopharyngeal carcinoma patients[J]. Exp Biol Med,2014,239(7):891-898.
[17] Katz B, Reis S T, Viana N I, et al. Comprehensive study of gene and microRNA expression related to epithelial-mesenchymal transition in prostate cancer[J]. PLoS one,2014,9(11):e113700.
[18] Tsukamoto O, Miura K, Mishima H, et al. Identification of endometrioid endometrial carcinoma-associated microRNAs in tissue and plasma[J]. Gynecol Oncol,2014,132(3):715-21.
[19] Sand M, Skrygan M, Georgas D, et al. Microarray analysis of microRNA expression in cutaneous squamous cell carcinoma[J]. J Dermatol Sci,2012,68(3):119-126.
[20] Suzuki A, Kusakai G A, Lu J, et al. ARK5 suppresses the cell death induced by nutrient starvation and death receptors via inhibition of caspase 8 activation, but not by chemotherapeutic agents or UV irradiation[J]. Oncogene,2003,22(40):6177-6182.
[21] Gen-Ichi K, Atsushi S, Tsutomu O, et al. ARK5 expression in colorectal cancer and its implications for tumor progression[J]. Am J Pathol,2004,164(3):987-995.
[22] Phippen N T, Bateman N W, Wang G, et al. NUAK1 (ARK5) Is Associated with Poor Prognosis in Ovarian Cancer[J]. Front Oncol. 2016,6:213. eCollection 2016.
[23] Chang X Z, Yu J, Liu H Y, et al. ARK5 is associated with the invasive and metastatic potential of human breast cancer cells[J]. J Cancer Res Clin Oncol,2012,138(2):247-254.
[24] Ye X T, Guo A J, Yin P F, et al. Overexpression of NUAK1 is associated with disease-free survival and overall survival in patients with gastric cancer[J]. Med Oncol,2014,31(7):61.
[25] Xu T, Zhang J, Chen W, et al. ARK5 promotes doxorubicin resistance in hepatocellular carcinoma via epithelial-mesenchymal transition[J]. Cancer Lett,2016,377(2):140-148.
[26] Lu S, Niu N, Guo H, et al. ARK5 promotes glioma cell invasion, and its elevated expression is correlated with poor clinical outcome[J]. Eur J Cancer,2013,49(3):752-763.
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