miR-363通过靶向HMGA2调控非小细胞肺癌的细胞生长与侵袭
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摘要
目的探讨miR-363通过靶向高迁移率蛋白A2(High mobility protein A1,HMGA2)调控非小细胞肺癌的细胞生长与侵袭行为及其作用机制。方法 qPCR检测非小细胞肺癌和癌旁正常组织、不同细胞株中miR-363的表达;分析miR-363的表达与非小细胞肺癌临床病理学参数之间的关系;CCK-8细胞增殖实验检测miR-363对非小细胞肺癌细胞增殖能力的影响;流式细胞术检测miR-363对非小细胞肺癌细胞周期的影响;划痕愈合实验和Transwell侵袭实验检测抑制miR-363后非小细胞肺癌细胞迁移和侵袭能力的变化;双荧光素酶报告基因检测miR-363与HMGA2的相互作用。结果与癌旁正常组织比较,非小细胞肺癌组织中miR-363表达明显增高;非小细胞肺癌细胞A549中miR-363的表达水平最高;抑制miR-363的表达后非小细胞肺癌细胞增殖能力减弱,促进其凋亡行为;抑制miR-363的表达后,A549细胞的迁移和侵袭能力受到相应的抑制;miR-363能与HMGA2的3’UTR特异性结合,调控HMGA2的表达活性。结论 miR-363在非小细胞肺癌的发生发展过程中起着重要作用,可通过靶向调节HMGA2的表达影响非小细胞肺癌细胞的生长和侵袭。
Objective To investigate the effect of miR-363 on the growth and invasion of non-small cell lung cancer cells by targeting HMGA2 and its mechanism.Methods qPCR was used to detect the expression of miR-363 in nonsmall cell lung cancer and adjacent normal tissues.The relationship between the expression of miR-363 and clinicopathological parameters of non-small cell lung cancer was analyzed.The proliferation of non-small cell lung cancer cells was detected by CCK-8cell proliferation assay.The effect of miR-363 on cell cycle of non-small cell lung cancer was detected by flow cytometry.Scratch healing assay and Transwell invasion assay were used to detect the migration and invasion ability of miR-363 cells after non-small cell lung cancer.Dual luciferase reporter assay detects the interaction of miR-363 with HMGA2.Results The expression of miR-363 in non-small cell lung cancer tissues was significantly higher than that in adjacent non-small cell lung cancer tissues and the highest expression level of miR-363 in non-small cell lung cancer A549 cells.The inhibitory effect of miR-363 on the proliferation of non-small cell lung cancer cells attenuated and promoted its apoptosis behavior.The inhibition of miR-363 expression inhibited the migration and invasion ability of A549 cells accordingly.miR-363 can specifically bind to the 3'UTR of HMGA2 and regulate the activity of HMGA2 expression.Conclusion miR-363 plays an important role in the development and progression of non-small cell lung cancer and may affect the growth and invasion of non-small cell lung cancer cells by regulating the expression of HMGA2.
Objective To investigate the effect of miR-363 on the growth and invasion of non-small cell lung cancer cells by targeting HMGA2 and its mechanism.Methods qPCR was used to detect the expression of miR-363 in nonsmall cell lung cancer and adjacent normal tissues.The relationship between the expression of miR-363 and clinicopathological parameters of non-small cell lung cancer was analyzed.The proliferation of non-small cell lung cancer cells was detected by CCK-8cell proliferation assay.The effect of miR-363 on cell cycle of non-small cell lung cancer was detected by flow cytometry.Scratch healing assay and Transwell invasion assay were used to detect the migration and invasion ability of miR-363 cells after non-small cell lung cancer.Dual luciferase reporter assay detects the interaction of miR-363 with HMGA2.Results The expression of miR-363 in non-small cell lung cancer tissues was significantly higher than that in adjacent non-small cell lung cancer tissues and the highest expression level of miR-363 in non-small cell lung cancer A549 cells.The inhibitory effect of miR-363 on the proliferation of non-small cell lung cancer cells attenuated and promoted its apoptosis behavior.The inhibition of miR-363 expression inhibited the migration and invasion ability of A549 cells accordingly.miR-363 can specifically bind to the 3'UTR of HMGA2 and regulate the activity of HMGA2 expression.Conclusion miR-363 plays an important role in the development and progression of non-small cell lung cancer and may affect the growth and invasion of non-small cell lung cancer cells by regulating the expression of HMGA2.
引文
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[3] Watanabe M,Kawaguchi T,Isa S,et al.Ultra-Sensitive Detection of the Pretreatment EGFR T790M Mutation in Non–Small Cell Lung Cancer Patients with an EGFR-Activating Mutation Using Droplet Digital PCR[J].Clinical Cancer Research,2015,21(15):3552-3560.
[4] Raaschou-Nielsen O,Andersen Z J,Beelen R,et al.Air pollution and lung cancer incidence in 17European cohorts:prospective analyses from the European Study of Cohorts for Air Pollution Effects(ESCAPE)[J].The lancet oncology,2013,14(9):813-822.
[5] Shaw A T,Kim D W,Nakagawa K,et al.Crizotinib versus chemotherapy in advanced ALK-positive lung cancer[J].New England Journal of Medicine,2013,368(25):2385-2394.
[6] Sozzi G,Boeri M,Rossi M,et al.Clinical utility of a plasmabased miRNA signature classifier within computed tomography lung cancer screening:a correlative MILD trial study[J].Journal of clinical oncology,2014,32(8):768-773.
[7] Yu T,Li J,Yan M,et al.MicroRNA-193a-3p and-5p suppress the metastasis of human non-small-cell lung cancer by downregulating the ERBB4/PIK3R3/mTOR/S6K2 signaling pathway[J].Oncogene,2015,34(4):413-423.
[8] Liu Z L,Wang H,Liu J,et al.MicroRNA-21(miR-21)expression promotes growth,metastasis,and chemo-or radioresistance in non-small cell lung cancer cells by targeting PTEN[J].Molecular and cellular biochemistry,2013,372(1-2):35-45.
[9] Zhao B,Han H,Chen J,et al.MicroRNA let-7cinhibits migration and invasion of human non-small cell lung cancer by targeting ITGB3and MAP4K3[J].Cancer letters,2014,342(1):43-51.
[10] Wang S H,Zhang W J,Wu X C,et al.The lncRNA MALAT1functions as a competing endogenous RNA to regulate MCL‐1expression by sponging miR‐363‐3p in gallbladder cancer[J].Journal of cellular and molecular medicine,2016,20(12):2299-2308.
[11] Hu F,Min J,Cao X,et al.MiR-363-3p inhibits the epithelialto-mesenchymal transition and suppresses metastasis in colorectal cancer by targeting Sox4[J].Biochemical and biophysical research communications,2016,474(1):35-42.
[12] Zhou P,Huang G,Zhao Y,et al.MicroRNA-363-mediated downregulation of S1PR1suppresses the proliferation of hepatocellular carcinoma cells[J].Cellular signalling,2014,26(6):1347-1354.
[13] Zhang R,Li Y,Dong X,et al.MiR-363sensitizes cisplatin-induced apoptosis targeting in Mcl-1in breast cancer[J].Medical Oncology,2014,31(12):347.
[14] Wang Y,Chen T,Huang H,et al.miR-363-3p inhibits tumor growth by targeting PCNA in lung adenocarcinoma[J].Oncotarget,2017,8(12):20133.
[15] Tsuji S,Kawasaki Y,Furukawa S,et al.The miR-363-GATA6-Lgr5pathway is critical for colorectal tumourigenesis[J].Nature communications,2014,5:3150.
[16] Fu W F,Chen W B,Dai L,et al.Inhibition of miR-141reverses cisplatin resistance in non-small cell lung cancer cells via upregulation of programmed cell death protein 4[J].Eur.Rev.Med.Pharm.Sci,2016,20:2565-2572.
[17] Hua J,He Y,Xu Y,et al.Emodin prevents intima thickness via Wnt4/Dvl-1/β-catenin signaling pathway mediated by miR-126in balloon-injured carotid artery rats[J].Experimental&molecular medicine,2015,47(6):e170.
[18] Zhao W,Wang Y,An Z,et al.Downregulation of miR-497promotes tumor growth and angiogenesis by targeting HDGF in non-small cell lung cancer[J].Biochemical and biophysical research communications,2013,435(3):466-471.
[19] Chen Y,Lu X,Wu B,et al.MicroRNA 363mediated positive regulation of c-myc translation affect prostate cancer development and progress[J].Neoplasma,2015,62(2):191-198.
[20] Zhang P F,Sheng L L,Wang G,et al.miR-363promotes proliferation and chemo-resistance of human gastric cancer via targeting of FBW7 ubiquitin ligase expression[J].Oncotarget,2016,7(23):35284.
[21] Wang S H,Zhang W J,Wu X C,et al.The lncRNA MALAT1functions as a competing endogenous RNA to regulate MCL‐1expression by sponging miR‐363‐3p in gallbladder cancer[J].Journal of cellular and molecular medicine,2016,20(12):2299-2308.
[22] Sun Q,Zhang J,Cao W,et al.Dysregulated miR-363affects head and neck cancer invasion and metastasis by targeting podoplanin[J].The international journal of biochemistry&cell biology,2013,45(3):513-520.
[23] Yang X,Zhao Q,Yin H,et al.MiR-33b-5p sensitizes gastric cancer cells to chemotherapy drugs via inhibiting HMGA2expression[J].Journal of Drug Targeting,2017,25(7):653-660.
[24] Kumar M S,Armenteros-Monterroso E,East P,et al.HMGA2functions as a competing endogenous RNA to promote lung cancer progression[J].Nature,2014,505(7482):212-217.