[1]鲍培龙,徐月亮,王孝彬.miR-153对肺腺癌细胞增殖、侵袭、迁移和凋亡的影响及机制研究[J].现代检验医学杂志,2022,37(01):107-113.[doi:10.3969/j.issn.1671-7414.2022.01.022]
　BAO Pei-long,XU Yue-liang,WANG Xiao-bin.Effects of miR-153 on Proliferation, Invasion, Metastasis and Apoptosis of Lung Adenocarcinoma Cells and Its Mechanism[J].Journal of Modern Laboratory Medicine,2022,37(01):107-113.[doi:10.3969/j.issn.1671-7414.2022.01.022]
点击复制

miR-153对肺腺癌细胞增殖、侵袭、迁移和凋亡的影响及机制研究()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年01期
页码:
107-113
栏目:
论　著
出版日期:
2022-01-15

文章信息/Info

Title:
Effects of miR-153 on Proliferation, Invasion, Metastasis and Apoptosis of Lung Adenocarcinoma Cells and Its Mechanism
文章编号:
1671-7414（2022）01-107-07
作者:
鲍培龙;  徐月亮;  王孝彬
（空军军医大学第二附属医院/ 唐都医院，西安 710038）
Author(s):
BAO Pei-long;  XU Yue-liang;  WANG Xiao-bin
（the Second Affiliated Hospital of Air Force Military Medical University /Tangdu Hospital, Xi’an 710038，China）
关键词:
miR-153; SRC蛋白; 肺腺癌; 增殖; 侵袭
分类号:
R734.2；R730.43
DOI:
10.3969/j.issn.1671-7414.2022.01.022
文献标志码:
A
摘要:
目的　探讨 miR-153对肺腺癌细胞增殖、迁移、侵袭及凋亡的影响及其相关作用机制。方法　构建 SRC-3’UTR-WT和 SRC-3’UTR-MUT载体，分别与 miR-153 mimic，mimic control共转染至肺腺癌 A549细胞，采用双荧光素酶报告基因实验验证 miR-153与 SRC的靶向关系。构建 miR-153过表达、敲低 miR-153和 SRC表达的 A549细胞系，采用 Western Blot检测 miR-153对 SRC蛋白表达的影响。通过 CCK-8法、细胞划痕实验、 Transwell侵袭实验及流式细胞仪分别检测 miR-153，SRC及 miR-153+SRC共转染对 A549细胞增殖、迁移、侵袭及凋亡的影响。结果　双荧光素酶报告基因实验证实 SRC是 miR-153的靶基因。 25例临床肺腺癌组织中 miR-153表达水平（ 13.251±4.256）较癌旁正常组织（ 25.312±6.527）显著降低（ t=7.739，P＜ 0.001），SRC表达水平（ 28.574±6.438）较癌旁正常组织（ 15.206±5.117）显著升高，差异有统计学意义 (t=8.128，P＜ 0.001)。随着临床分期和病理学分级的增高，肺腺癌组织中 miR-153表达逐渐降低（ F=13.351，8.479，P<0.01），SRC表达逐渐升高（ F=9.812，10.521，P<0.001），差异均有统计学意义。肺腺癌组织中 miR-153与 SRC表达呈显著负相关 (r=-0.726，P<0.05)。过表达 miR-153显著抑制 SRC蛋白表达（ t=7.075， P=0.002），而降低 miR-153 表达得到与之相反的结果。过表达 miR-153和敲低 SRC蛋白表达显著抑制 A549细胞的增殖（t=22.265，21.783，均 P<0.001）、迁移（ t=7.287，4.819，P=0.002，0.009）及侵袭（ t=10.043，10.563，P=0.001，
Abstract:
Objective　To investigate the effects of miR-153 on proliferation, migration, invasion and apoptosis of lungadenocarcinoma cells and its related mechanisms. Methods　SRC-3’UTR-WT and SRC-3’ UTR-MUT vectors were constructedand co-transfected into lung adenocarcinoma A549 cells with miR-153 mimic and mimic control, respectively. Dual-luciferasereporter gene experiment was used to verify the targeted relationship between miR-153 and SRC. A549 cell lines with miR-153overexpression and knockdown of miR-153 and SRC expression were constructed, and the effect of miR-153 on the expressionof SRC protein was detected by Western Blot. The effects of miR-153, SRC and miR-153 +SRC cotransfection on theproliferation, migration, invasion and apoptosis of A549 cells were detected by CCK-8 method, cell scratch test, Transwellinvasion test and flow cytometry, respectively. Results　SRC was confirmed to be the target gene of miR-153 by dual luciferasereporter gene assay. The expression level of miR-153 in 25 clinical lung adenocarcinoma tissues (13.251±4.256) wassignificantly lower than that in adjacent normal tissues (25.312±6.527), (t=7.739, P< 0.001), SRC expression level(28.574±6.438) was significantly higher than that of adjacent normal tissues (15.206±5.117), the differences was statisticallysignificant (t=8.128, P< 0.001). With the increase of clinical stage and pathological grade, the expression of miR-153 in lungadenocarcinoma tissues decreased gradually (F=13.351, 8.479, P<0.01), and the expression of SRC increased gradually, thedifferences was statistically significant(F=9.812, 10.521, P<0.001). There was a significant negative correlation between miR-153and SRC expression in lung adenocarcinoma tissues (r=-0.726, P<0.05). Overexpression of miR-153 significantly inhibitedthe expression of SRC protein (t=7.075, P=0.002), while decreasing the expression of miR-153 resulted in the opposite result.Overexpression of miR-153 and knockdown of SRC protein significantly inhibited the proliferation of A549 cells (t=22.265,21.783, all P<0.001), migration (t=7.287, 4.819, P=0.002, 0.009) and invasion (t=10.043, 10.563, all P=0.001), promoting cellapoptosis (t=3.918, 6.735, P=0.017, 0.003). Inhibition of miR-153 expression resulted in the opposite result. Co-expression ofmiR-153+ SRC reversed the inhibition/promotion effect of overexpression of miR-153 on proliferation, migration and apoptosisof lung adenocarcinoma cells. Conclusion　Mir-153 was significantly under-expressed in lung adenocarcinoma, and it couldinhibit the proliferation, invasion and metastasis of lung adenocarcinoma cells and promote cell apoptosis by targeting down theexpression of SRC.

参考文献/References:

[1] 刘宗超，李哲轩，张阳，等．2020 全球癌症统计报告解读[J]. 肿瘤综合治疗电子杂志, 2021,7(2):1-13.LIU Zongchao,LI Zhexuan,ZHANG Yang,et al.Interpretation on the report of global cancer statistics2020[J]. Journal of Multidisciplinary CancerManagement(Electronic Version), 2021,7(2):1-13.
[2] ANASTASIADOU E, JACOB L S, SLACK F J. NoncodingRNA networks in cancer[J]. Nature ReviewsCancer, 2018, 18(1): 5-18.
[3] 席俊峰, 彭彦才, 马兴聪, 等．Aurora-B 激酶的表达与非小细胞肺癌患者预后及耐药的相关性分析[J].现代检验医学杂志,2020,35(6):59-63.XI Junfeng, PENG Yancai, MA Xingcong, et al.Study on the relationship between high expression ofAurora-B kinase and drug resistance and prognosis inpatients with non-small cell lung cancer [J]. Journal ofModern Laboratory Medicine,2020,35(6):59-63.
[4] TONG Zhan, CUI Qinghua, WANG Juan, et al.TransmiR v2.0: an updated transcription factormicroRNAregulation database[J]. Nucleic AcidsResearch, 2019, 47(D1) : D253-D258.
[5] 黄刚, 陈霏, 肇玉博, 等. 非小细胞肺癌患者血浆miRNA-145 和miRNA-221 表达与临床特征及术后复发的相关性研究[J]. 现代检验医学杂志, 2019, 34(4):40-44.HUANG Gang, CHEN Fei, ZHAO Yubo, et al.Study on the correlation between the expression ofmicroRNA145 and microRNA221 in plasma andclinical characteristics and postoperative recurrence inpatients with non-small cell lung cancer [J]. Journal ofModern Laboratory Medicine,2019,34(4):40-44.
[6] HOSSEINAHLI N, AGHAPOUR M, DUIJF P H G.Treating cancer with microRNA replacement therapy:A literature review[J]. Journal of Cellular Physiology,2018, 233(8): 5574-5588.
[7] WANG Dongchang, WANG Lingchan , ZHANG Yu,et al. PYCR1 promotes the progression of non-smallcelllung cancer under the negative regulation of miR-488[J]. Biomedicine & Pharmacotherapy, 2019, 111:588-595.
[8] 谢琼, 胡桂明, 王洪涛.miRNA-223-3 p 调控JAK2/STAT3 信号通路对胃癌细胞增殖、凋亡、迁移的影响[J]. 郑州大学学报( 医学版)，2018, 53(5): 629-634.XIE Qiong, HU Guiming, WANG Hongtao, etal. Effect of miRNA-223-3p by regulation ofJAK2/STAT3 signaling pathway on proliferation,apoptosis and migration of gastric cancer cells[J]. Journal of Zhengzhou University(MedicalSciences),2018,53(5):629-634.
[9] 刘卜玮，蔡明成，杨雪，等.microRNA 调控细胞凋亡的研究进展[J]. 生理科学进展,2018,49(4):309-314.LIU Buwei, CAI Mingcheng, YANG Xue, et al. Researchprogress of microRNA regulation of apoptosis[J].Progress in Physiological Sciences,2018,49(4):309-314.
[10] HUANG Mengqin, LI Chengfa, KONG Fanliang, etal. Prognostic and clinicopathological significance ofMicroRNA-153 in human cancers: A meta-analysis[J].Medicine, 2020, 99(46): e22833.
[11] CUI Yixin, SUN Gongqin . Structural versatility thatserves the function of the HRD motif in the catalyticloop of protein tyrosine kinase, Src[J]. Protein Science,2019, 28(3): 533-542.
[12] 郭冬梅, 谢莉莉, 谢奇.Src 激酶抑制剂PP2 对人舌鳞癌Tca8113 细胞侵袭和转移的抑制作用[J]. 西安交通大学学报( 医学版) ,2019,40(2):290-293.GUO Dongmei, XIE Lili, XIE Qi. Inhibition ofinvasion and metastasis of human tonguesquamouscellcarcinoma Tca8113 cells by Src kinase inhibitorPP2 [J]. Journal of Xi’an Jiaotong University(MedicalSciences),2019,40(2):290-293.
[13] 王登嵘，刘显，肖健．Src 蛋白激酶在疾病中的功能及研究现状[J]. 医学与哲学,2018,39(1):58-60,79.WANG Dengrong, LIU Xian, XIAO Jian. The functionand research status of src protein kinase in diseases [J].Medicine & Philosophy,2018,39(1):58-60,79.
[14] 王琴，刘力，乐意，等．Src 激酶抑制剂研究进展[J].中国药物化学杂志,2021,31(4):312-319.WANG Qin, LIU Li, LE Yi, et al. The progress of Srcinhibitors [J]. Chinese Journal of Medicinal Chemistry,2021,31(4):312-319.
[15] LIAO Zhicong, WANG Xiaojun, LIANG Hongwei, etal. MiR-1 suppresses the proliferation and promotes theapoptosis of esophageal carcinoma cells by targetingSrc[J]. Cancer Medicine 2017, 6(12) : 2957-2965.
[16] LIU Yennien, YIN Juanjuan, BARRETT B, et al. Lossof androgen-regulated microRNA 1 activates SRC andpromotes prostate cancer bone metastasis[J]. Molecularand Cellular Biology, 2015, 35(11): 1940-1951.
[17] KLOBU?AR M, SEDI? M, GEHRIG P, et al.Basement membrane protein ladinin-1 and the MIFCD44-β1 integrin signaling axis are implicatedin laryngeal cancer metastasis[J]. Biochimica etBiophysica Acta, 2016, 1862(10): 1938-1954.
[18] YASMIN S, HAQUE W S,ISLAM S M J,et al. Roleof immunohistochemistry in cell block sections forcategorization of non-small cell carcinoma of lung[J].BIRDEM Medical Journal, 2019, 9(3): 223-228.
[19] WANG Liyan, LI Xiaonan, ZHANG Wei, et al. miR24-2 promotes malignant progression of human livercancer stem cells by enhancing tyrosine kinase Srcepigenetically[J]. Molecular Therapy, 2020, 28(2): 572-586.
[20] JIANG Guojun, LIU Yaming, WANG Ru, et al.Activation of Src mediates acquired cisplatin resistancein human lung carcinoma cells[J]. Anti-Cancer Drugs,2020, 31(2): 123-130.
[21] 周小敏, 赵展庆, 林奇栋, 等. 微小RNA-423 靶向调控BAP1 表达及对肺鳞癌细胞增殖和凋亡的影响[J]. 临床肿瘤学杂志,2019,24(8):678-683.ZHOU Xiaomin, ZHAO Zhanqing, LIN Qidong, etal. Targeted regulation of microRNA-423 on BAP1expression and its effect on proliferation and apoptosisof lung squamous cell carcinoma cells [J]. ChineseClinical Oncology,2019,24(8):678-683.
[22] ZHAO Guoli, ZHANG Yueying, ZHAO Zhonghua, etal. MiR-153 reduces stem cell-like phenotype and tumorgrowth of lung adenocarcinoma by targeting Jagged1[J].Stem Cell Research & Therapy, 2020, 11(1): 170.
[23] WANG Heping, YU Yanzhang, FAN Shuxin, et al.Knockdown of long noncoding RNA TUG1 inhibits theproliferation and cellular invasion of osteosarcoma cellsby sponging miR-153[J]. Oncology Research, 2018,26(5): 665-673.
[24] 李冠军, 亚国伟, 唐正严, 等．miR-153 靶向PRDM2基因并通过JAK/STAT 信号通路影响膀胱癌的侵袭和迁移[J]. 中国病理生理杂志, 2018, 34 (1): 58-63.LI Guanjun, YA Guowei, TANG Zhengyan, et al. Effectof miR-153-targeted PRDM2 on invasion and migrationabilities of bladder cancer cells by JAK/STAT signalingpathway [J]. Chinese Journal of Pathophysiology, 2018,34(1):58-63.
[25] SHI Dongliang, LI Yong, FAN Liqiao, et al.Upregulation of miR-153 inhibits Triple-Negativebreast cancer progression by targeting ZEB2-MediatedEMT and contributes to better prognosis[J]. OncoTargets and Therapy, 2019, 12: 9611-9625.
[26] DENG Xianzhao, GUO Bomin, FAN Yonben. MiR-153-3p suppresses cell proliferation, invasion andglycolysis of thyroid cancer through inhibiting E3F3expression[J]. Onco Targets and Therapy, 2021, 14:519-529.
[27] SHI Fei,SHI Zhonghua,ZHAO Yaodong,et al. CircRNAhsa-circ-0014359 promotes glioma progression byregulating miR-153/PI3K signaling[J]. Biochemical andBiophysical Research Communications, 2019, 510(4):614-620.
[28] DJEUNGOUE-PETGA M, LURETTE O, JEAN S, etal. Intramitochondrial Src kinase links mitochondrialdysfunctions and aggressiveness of breast cancercells[J]. Cell Death & Disease, 2019, 10(12): 940.
[29] KIM E S, CHOI J Y, HWANG S J , e t a l .Hypermethylation of miR-205-5p by IR governsaggressiveness and metastasis via regulating Bcl-w andSrc[J]. Molecular Therapy Nucleic Acids, 2019, 14:450-464.
[30] CANER A , ASIK E, OZPOLAT B. SRC Signaling inCancer and Tumor Microenvironment[M].// Advancesin Experimental Medicine and Biology 2020: 1270,InTumor Microenvironment : 57-71.

备注/Memo

备注/Memo:
基金项目：陕西省重点研发计划项目（2019SF-061）。
作者简介：鲍培龙（1989-），男，硕士研究生，主治医师，研究方向：胸部肿瘤的微创治疗，E-mail：qianfa815@yeah.net。
通讯作者：王孝彬，研究方向：胸部肿瘤的诊治。
更新日期/Last Update: 1900-01-01