miR-485-3p regulates radiosensitivity of gastric cancer cells by targeting TLR1/NF-κB signaling pathway
Li Mingjun,Wu Guangyin,Fan Huijie,Gu Hao,Fan Ruitai,Shi Yonggang,Zhang Mingzhi
Department of Medical Oncology (Li MJ, Fan HJ, Zhang MZ),Department of Radiotherapy (Gu H,Fan RT,Shi YG),First Affiliated Hospital of Zhengzhou University,Zhengzhou,450052 China;Department of Radiotherapy,Henan Provincial People’s Hospital (Wu GY)
Abstract：Objective To investigate whether miR-485-3p plays a role in regulation of radiosensitivity of gastric cancer cells by targeting TLR1. Methods Quantitative real-time PCR and Western blot were used to determine the expression of miR-485-3p and TLR1, respectively. The interaction between miR-485-3p and TLR1 was verified by target prediction software (DIANA, TargetScan, and miRanda) and dual luciferase reporter assay. Gastric cancer MGC803 cells transfected with miR-485-3p mimic or TLR1 siRNA were exposed to irradiation. Apoptosis assay, colony formation assay, and MTT assay were used to evaluate the changes in radiosensitivity of gastric cancer cells. Dual luciferase reporter assay was used to determine the effects of miR-485-3p overexpression and TLR1 silencing on the activity of NF-κB. Western blot was used to study the effects of miR-485-3p overexpression and TLR1 silencing on NF-κB target genes. Results In gastric cancer cells exposed to radiation, the expression of miR-485-3p was downregulated and the expression of TLR1 was upregulated. TLR1 was predicted to be the target of miR-485-3p by target prediction software. Dual luciferase reporter assay further confirmed TLR1 as the direct target of miR-485-3p. miR-485-3p negatively regulated the expression of TLR1. The overexpression of miR-485-3p, as well as TLR1 silencing, increased the apoptosis rate of cells, reduced colony formation and cell proliferation, and enhanced the radiosensitivity of the cells. Both miR-485-3p overexpression and TLR1 silencing reduced the activity of NF-κB and downregulated the expression of multiple NF-κB target genes. Conclusions miR-485-3p enhances the radiosensitivity of gastric cancer cells probably by targeting TLR1 and regulating the NF-κB signaling pathway.
Li Mingjun,Wu Guangyin,Fan Huijie et al. miR-485-3p regulates radiosensitivity of gastric cancer cells by targeting TLR1/NF-κB signaling pathway[J]. Chinese Journal of Radiation Oncology, 2016, 25(8): 879-884.
[1] Yang H,Wang B,Yan J,et al. Toll-like receptor 2 promotes invasion by SGC-7901 human gastric carcinoma cells and is associated with gastric carcinoma metastasis[J].Ann Clin Lab Sci,2014,44(2):158-166 [2] Weber ANR,Frsti A.Toll-like receptor genetic variants and colorectal cancer[J].Oncoimmunology,2014,3(2):e27763.DOI:10.4161/onci.27763 [3] Voo KS,Foglietta M,Percivalle E,et al. Baladandayuthapani,Selective targeting of Toll-like receptors and OX40 inhibit regulatory T–cell function in follicular lymphoma[J].Int J Cancer,2014,135(12):2834-2846.DOI:10.1002/ijc.28937 [4] Vacchelli E,Eggermont A,Sautès-Fridman C,et al. Trial Watch:toll-like receptor agonists for cancer therapy[J].Oncoimmunology,2013,2(8):e25238.DOI:10.4161/onci.25238 [5] Lan FM,Yue X,Ren G,et al.miR-15a/16 enhances radiation sensitivity of non-small cell lung cancer cells by targeting the TLR1/NF-κB signaling pathway[J].Int J Radiat Oncol Biol Phys,2015,91(1):73-81.DOI:10.1016/j.ijrobp.2014.09.021 [6] He J,Hua J,Ding N,et al. Modulation of microRNAs by ionizing radiation in human gastric cancer[J].Oncol Rep,2014,32(2):787-793.DOI:10.3892/or.2014.3246 [7] Ueda T,Volinia S,Okumura H,et al. Relation between microRNA expression and progression and prognosis of gastric cancer:a microRNA expression analysis[J].Lancet Oncol,2010,11(2):136-146.DOI:10.1016/S1470-2045(09)70343-2 [8] Guo JM,Miao Y,Xiao BX,et al. Differential expression of microRNA species in human gastric cancer versus non-tumorous tissues[J].J Gastroenterol Hepatol,2009,24(4):652-657.DOI:10.1111/j.1440-1746.2008.05666.x [9] Liu R,Zhang CN,Hu ZB,et al. A five-microRNA signature identified from genome-wide serum microRNA expression profiling serves as a fingerprint for gastric cancer diagnosis[J].Eur J Cancer,2011,47(5):784-791.DOI:10.1016/j.ejca.2010.10.025 [10] Wang HJ,Ruan HJ,He XJ,et al. MicroRNA-101 is down-regulated in gastric cancer and involved in cell migration and invasion[J].Eur J Cancer,2010,46(12):2295-2303.DOI:10.1016/j.ejca.2010.05.012 [11] Ding L,Xu YJ,Zhang W,et al. MiR-375 frequently downregulated in gastric cancer inhibits cell proliferation by targeting JAK2[J].Cell Res,2010,20(7):784-793.DOI:10.1038/cr.2010.79 [12] Tie J,Pan YL,Zhao LN,et al. MiR-218 inhibits invasion and metastasis of gastric cancer by targeting the Robo1 receptor[J].PLoS Genet,2010,6(3):e1000879.DOI:10.1371/journal.pgen.1000879 [13] Liu YX,Xing R,Zhang XD,et al.miR-375 targets the p53 gene to regulate cellular response to ionizing radiation and etoposide in gastric cancer cells[J].DNA Repair,2013,12(9):741-750.DOI:10.1016/j.dnarep.2013.06.002 [14] Zhang CZ,Han L,Zhang AL,et al. MicroRNA-221 and microRNA-222 regulate gastric carcinoma cell proliferation and radioresistance by targeting PTEN[J].BMC Cancer,2010,10:367.DOI:10.1186/1471-2407-10-367 [15] Formosa A,Markert EK,Lena AM,et al. MicroRNAs,miR-154,miR-299-5p,miR-376a,miR-376c,miR-377,miR-381,miR-487b,miR-485-3p,miR-495 and miR-654-3p,mapped to the 14q32.31 locus,regulate proliferation,apoptosis,migration and invasion in metastatic prostate cancer cells[J].Oncogene,2014,33(44):5173-5182.DOI:10.1038/onc.2013.451 [16] Anaya-Ruiz M,Bandala C,Perez-Santos JLM.miR-485 acts as a tumor suppressor by inhibiting cell growth and migration in breast carcinoma T47D cells[J].Asian Pac J Cancer Prev,2013,14(6):3757-3760.DOI:10.7314/APJCP.2013.14.6.3757 [17] 路钊,郑少鹏,牛静,等.BATF2/SARI通过抑制p53依赖的NF-κB转录活性诱导肿瘤细胞凋亡[J].中国生物化学与分子生物学报,2011,27(6):524-532 Lu Z,Zheng SP,Niu J,et al. BATF2/SARI induces tumor cell apoptosis by inhibiting p53-dependent NF-κB activity[J].Chin J Biochem Mol Biol,2011,27(6):524-532 [18] 夏永华,刘冬,张彩凤,等.NF-κB信号通路的阻断对皮肤鳞癌SCL-1细胞凋亡的影响[J].北京大学学报(医学版),2011,43(2):179-182.DOI:10.3969/j.issn.1671-167X.2011.02.004 Xia YH,Liu D,Zhang CF,et al. Effect of blockade of NF-κB signaling pathway on cell apoptosis in cutaneous squamous cell carcinoma[J].J Peking Univ (Health Sci),2011,43(2):179-182.DOI:10.3969/j.issn.1671-167X.2011.02.004 [19] 肖雪飞,杨明施,孙圣华.姜黄素对脓毒症急性肺损伤的炎症反应及NF-κB信号通路的影响[J].中国现代医学杂志,2013,23(30):19-22 Xiao XF,Yang MS,Sun SH.Effect of curcumin on inflammation and NF-κB signaling pathway in septic acute lung injury rats[J].China J Mod Med,2013,23(30):19-22
2016, Vol. 25 
