Inhibitory effect of microRNA-149-5p in the proliferation and migration of HepG2 and Bel-7402 hepatoma cells
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摘要: 目的明确microRNA-149-5p(miR-149-5p)在肝癌组织中的表达,并探讨其在肝癌细胞中的分子生物学作用。方法收集2010年1月-2014年1月内蒙古医科大学附属医院通过肝切除术获得的65例肝癌组织及相应癌旁组织,采用荧光定量PCR检测65例肝癌组织及相应癌旁组织中miR-149-5p的表达情况。细胞实验分2组,实验组转染miR-149-5p模拟物,对照组转染模拟物阴性对照,采用噻唑蓝比色法和创伤愈合试验检测miR-149-5p对Hep G2和Bel-7402细胞增殖及迁移能力的影响。计量资料组间比较采用t检验或单因素方差分析。结果 miR-149-5p在肝癌组织的表达量(0.14±0.06)明显低于配对的癌旁组织(2.56±0.42),两者差异有统计学意义(t=7.79,P<0.05);miR-149-5p在肝癌细胞Hep G2、Bel-7402和正常肝上皮细胞L02中的表达量分别为(1.43±0.25)、(1.77±0.32)和(5.68±0.74),3组间比较差异有统计学意义(F=11.27,P<0.05)。体外功能实验发现,miR-149-5p模拟物可...Abstract: Objective To investigate the expression of microRNA-149-5p (miR-149-5p) in liver cancer tissue and its molecular and biological role in hepatoma cells. Methods A total of 65 liver cancer tissue samples and corresponding adjacent tissue samples were collected from January 2010 to January 2014, in the Affiliated Hospital of Inner Mongolia Medical University. Quantitative real-time PCR was used to measure the expression of miR-149-5p in liver cancer tissue and corresponding adjacent tissue. The cells were divided into two groups; the cells in the experimental group were transfected with miR-149-5p mimic, and those in the control group were transfected with the negative control of the mimic. MTT colorimetry and wound-healing assay were performed to determine the effect of miR-149-5p on the proliferation and migration of HepG2 and Bel-7402 cells. The t-test or a one-way analysis of variance was used for comparison of continuous data between groups. Results The liver cancer tissue had significantly lower expression of miR-149-5p than the adjacent tissue (0. 14 ± 0. 06 vs 2. 56 ± 0. 42, t = 7. 79, P < 0. 05) . There were significantly differences in the expression of miR-149-5p in Hep G2 (1. 43 ± 0. 25) 、Bel-7402 (1. 77 ± 0. 32) , and the normal hepatic epithelial cells (5. 68 ± 0. 74) (F = 11. 27, P < 0. 05) . The in vitro functional experiment showed that miR-149-5p mimic significantly inhibited the proliferation of 24、48、72 hour of Hep G2 and Bel-7402 hepatoma cells (Hep G2: t = 4. 98, 5. 17, 7. 78, all P < 0. 05; Bel-7402: t = 6. 83, 7. 09, 15. 67, all P < 0. 05) and inhibited the migration of Hep G2 and Bel-7402 hepatoma cells (t = 23. 11, 17. 42, both P < 0. 05) . Conclusion The expression of miR-149-5p is downregulated in liver cancer tissue, and overexpressed miR-149-5p can inhibit the proliferation and migration of hepatoma cells, suggesting that miR-149-5p may be a promising and effective molecular target for the genetic treatment of liver cancer.
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Key words:
- liver neoplasms, experimental /
- microRNAs /
- cell proliferation /
- cell movement
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[1]SIEGEL RL, MILLER KD, JEMAL A.Cancer Statistics, 2016[J].CA Cancer J Clin, 2016, 66 (1) :7-30. [2]HUANG J.Current progress in epigenetic research for hepatocarcinomagenesis[J].Sci China C Life Sci, 2009, 52 (1) :31-42. [3]ZHAO LJ, LI ZF, SHI RY, et al.Expression of NOB1 in liver cancer tissues and its significance[J].J Clin Hepatol, 2016, 32 (4) :739-741. (in Chinese) .赵丽娟, 李志锋, 石荣亚, 等.NOB1在肝癌组织中的表达及意义[J].临床肝胆病杂志, 2016, 32 (4) :739-741. [4]BARTEL DP.Micrornas:genomics, biogenesis, mechanism, and function[J].Cell, 2004, 116 (2) :281-297. [5]PASQUINELLI AE.Micrornas and their targets:recognition, regulation and an emerging reciprocal relationship[J].Nat Rev Genet, 2012, 13 (4) :271-282. [6]HWANG HW, MENDELL JT.Micrornas in cell proliferation, cell death, and tumorigenesis[J].Br J Cancer, 2006, 94 (6) :776-780. [7]SCHICKEL R, BOYERINAS B, PARK SM, et al.Micrornas:key players in the immune system, differentiation, tumorigenesis and cell death[J].Oncogene, 2008, 27 (45) :5959-5974. [8]MENDELL JT.Micrornas:critical regulators of development, cellular physiology and malignancy[J].Cell Cycle, 2005, 4 (9) :1179-1184. [9]LI ZW, WU T, FU YX, et al.Expression and roles of MicroRNA-31 in HCC[J].Chin J Clin Pharmacol Ther, 2015, 20 (10) :1098-1101. (in Chinese) 李中文, 吴涛, 付应霄, 等.MicroRNA-31在肝癌中的表达和作用[J].中国临床药理学与治疗学, 2015, 20 (10) :1098-1101. [10]JIN L, LI Y, LIU J, et al.Tumor suppressor mir-149-5p is associated with cellular migration, proliferation and apoptosis in renal cell carcinoma[J].Mol Med Rep, 2016, 13 (6) :5386-5392. [11]XU K, LIU X, MAO X, et al.Microrna-149 suppresses colorectal cancer cell migration and invasion by directly targeting forkhead box transcription factor foxm1[J].Cell Physiol Biochem, 2015, 35 (2) :499-515. [12]CHAN SH, HUANG WC, CHANG JW, et al.Microrna-149 targets git1 to suppress integrin signaling and breast cancer metastasis[J].Oncogene, 2014, 33 (36) :4496-4507. [13] SCHMITTGEN TD, LIVAK KJ.Analyzing Real-Time PCR data by the comparative C (T) method[J].Nat Protoc, 2008, 3 (6) :1101-1108. [14]WONG CM, NG IO.Molecular pathogenesis of hepatocellular carcinoma[J].Liver Int, 2008, 28 (2) :160-174. [15]MORENO-MOYA JM, VILELLA F, SIMON C.Microrna:key gene expression regulators[J].Fertil Steril, 2014, 101 (6) :1516-1523. [16]CALIN GA, CROCE CM.Microrna signatures in human cancers[J].Nat Rev Cancer, 2006, 6 (11) :857-866. [17]MEDINA PP, SLACK FJ.Micrornas and cancer:an overview[J].Cell Cycle, 2008, 7 (16) :2485-2492. [18]NIKITINA EG, URAZOVA LN, STEGNY VN.Micrornas and human cancer[J].Exp Oncol, 2012, 34 (1) :2-8. [19]FARAZI TA, HOELL JI, MOROZOV P, et al.Micrornas in human cancer[J].Adv Exp Med Biol, 2013, 774:1-20. [20]SHENOUDA SK, ALAHARI SK.Microrna function in cancer:oncogene or a tumor suppressor?[J].Cancer Metastasis Rev, 2009, 28 (3-4) :369-378. [21]FENG M, E CY, LI H, et al.Expression of miR-196a in liver cancer cells and its promotion effect on proliferation[J].J Jilin Univ:Med Edit, 2015, 41 (1) :120-124. (in Chinese) 冯默, 鄂长勇, 李航, 等.MiR-196a在肝细胞癌中的表达及其促增殖作用[J].吉林大学学报:医学版, 2015, 41 (1) :120-124. [22]NANA-SINKAM SP, CROCE CM.Micrornas as therapeutic targets in cancer[J].Transl Res, 2011, 157 (4) :216-225.
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