miR-625-3p在结肠癌组织和细胞中的表达

目的:探讨微小RNA(mi R)-625-3p在结肠癌组织和细胞中的表达,了解其作用机制。方法:利用q RT-PCR分析结肠癌细胞株、癌组织和癌旁组织中mi R-625-3p的表达水平,探讨mi R-625-3p表达水平与结肠癌临床病理参数之间的关系;利用碘化丙啶染色和流式细胞术检测细胞凋亡和细胞周期变化;Western blot法检测mi R-625-3p对结肠癌细胞株凋亡相关蛋白的影响,初步了解其发挥影响的可能机制。结果:结肠癌组织mi R-625-3p的表达量比癌旁组织高(P0.05),mi R-625-3p的表达量与结肠癌浸润深度、TNM分期及有无远处转移关系密切(P0.05)。mi R-625-3p在结肠癌SW620细胞中的表达水平高于在SW480细胞中的表达。抑制mi R-625-3p的表达能显著抑制结肠癌细胞的周期(P0.05)和促进凋亡;转染mi R-625-3p后,Bcl-2的表达量增加(P0.05),Bax表达量没有显著变化。结论:mi R-625-3p能促进结肠癌细胞增殖,抑制结肠癌细胞凋亡,可能是结肠癌新的抗癌靶点。     

LncRNA OGFRP1 promotes angiogenesis and epithelial-mesenchymal transition in colorectal cancer cells through miR-423-5p/CTCF axis

ObjectiveTo investigate the mechanism of lncRNA OGFRP1 affecting angiogenesis and epithelial-mesenchymal transition (EMT) in colorectal cancer (CRC) and provide a new target for the treatment of CRC.MethodsThe expressions of OGFRP1, miR-423-5p, and CTCF were measured in CRC cell lines (HT29, LoVo, HCT116, SW620, and SW480) and normal colonic epithelial cells NCM460. Gain and loss of function experiments were performed on HCT116 and SW620 cells, after which the proliferation, apoptosis, EMT, invasion, and migration of the cells were measured using CCK-8 and colony formation assays, flow cytometry, Western blotting, Transwell, and scratch assay. The transfected cells were incubated with human umbilical vein endothelial cells (HUVECs) to assess angiogenesis using tube formation assay. ELISA was performed to detect VEGF in the conditioned medium of HCT116 and SW620 cells. The interactions among OGFRP1, CTCF and miR-423-5p were validated by dual-luciferase reporter assay.ResultsCRC cell lines had increased expression levels of OGFRP1 and CTCF and a suppressed expression level of miR-423-5p when compared with NCM460 cells. Suppression on OGFRP1 or CTCF and overexpression of miR-423-5p led to inhibited proliferation, EMT, invasion and migration and increased apoptosis of HCT116 and SW620 cells. HUVECs incubated with cells transfected with si-OGFRP1, si-CTCF or miR-423-5p mimic had suppressed angiogenesis ability. The effect of OGFRP1 suppression in CRC cells could be counteracted by miR-423-5p inhibition. Both CTCF and OGFRP1 could bind to miR-423-5p.ConclusionOGFRP1 promotes proliferation, EMT, and angiogenesis in CRC through miR-423-5p/CTCF axis.     

miR-424-5p promotes the proliferation and metastasis of colorectal cancer by directly targeting SCN4B

BackgroundColorectal cancer (CRC) is one of the most common malignancies worldwide usually diagnosed at advanced stages which causes poor prognosis of patients. Therefore, novel diagnostic biomarkers and therapeutic targets are urgently needed.Materials and methodsmiR-424-5p was identified through integrated analysis of three public databases. Loss-of-function experiments in HT29 and SW480 cells and mouse xenograft models were performed to explore the regulatory role of miR-424-5p in CRC. Bioinformatics analysis was used for predicting targets of miR-424-5p and its functional and pathway enrichment analysis.ResultsmiR-424-5p expression was significantly upregulated in CRC tissues and cell lines and associated with prognosis of CRC patients. Experiments in vitro and in vivo showed miR-424-5p promotes CRC cell proliferation and metastasis by directly inhibiting SCN4B. Besides, CRC cells secret miR-424-5p into peripheral blood through exosomes and circulating exosomal miR-424-5p could discriminate CRC patients with early stage from healthy people with AUC value of 0.82.ConclusionsmiR-424-5p serves as an oncogene in CRC and circulating exosomal miR-424-5p is a novel potential diagnostic biomarker of CRC patients.     

miR-23a和ESRP1在直肠癌中的作用

目的:研究miR-23a和上皮剪接调节蛋白1(epithelial splicing regulatory protein 1,ESRP1)在直肠癌组织及细胞系中的表达,以及对体外直肠癌细胞活力和凋亡的作用。方法:采用RT-q PCR分析miR-23a在36例直肠癌组织和癌旁组织中的表达,免疫组化检测ESRP1在直肠癌组织中的表达,分析miR-23a和ESRPl在直肠癌组织中的相关性;利用RT-q PCR检测miR-23a在直肠癌Caco-2和SW480细胞及人正常结肠上皮细胞株NCM460中的表达;合成miR-23a inhibitor和inhibitor阴性对照(inhibitor NC),并将其分别转染至SW480细胞后,通过CCK-8法检测miR-23a inhibitor转染SW480细胞后对细胞活力的影响,流式细胞术检测转染后细胞凋亡率,Transwell小室实验检测细胞侵袭;通过Western blot技术检测SW480细胞中ESRPl蛋白的表达;构建野生型pGL3-ESRP1-3’UTR(wt-pGL3-ESRP1-3’UTR)或突变型pGL3-ESRP1-3’UTR(mut-pGL3-ESRP1-3’UTR)质粒,并分别与miR-23a inhibitor或inhibitor NC共转染至HEK293和SW480细胞中,利用双萤光素酶报告基因检测试剂盒说明检测双萤光素酶活性;将ESRP1 mimic或mimic NC瞬时转染SW480细胞后,CCK-8法和流式细胞术分别检测细胞活力和凋亡;Western blot法检测瞬转ESRP1 mimic后对ESRP1、caspase-3、Smac和XIAP蛋白表达的影响。结果:miR-23a和ESRP1在直肠癌组织的表达较癌旁正常组织分别上调和下调,两者呈明显负相关(P0.01);miR-23a的表达与直肠癌的淋巴结转移和肿瘤浸润深度相关;与NCM460细胞相比较,miR-23a在SW480细胞中的表达量显著上调(P0.01);转染miR-23a inhibitor后,SW480细胞活力较inhibitor NC组显著下降(P0.01);转染miR-23a inhibitor后SW480细胞早期凋亡率明显升高,同时细胞体外侵袭能力受到抑制;萤光素酶报告基因结果表明ESRP1是miR-23a的直接靶基因;转染miR-23a inhibitor至SW480细胞后ESRP1蛋白表达水平明显升高;ESRP1 mimic转染SW480细胞后可抑制细胞活力并诱导细胞凋亡,同时上调caspase-3和Smac的表达,下调XIAP的表达。结论:miR-23a可通过负向调控下游靶基因ESRP1从而影响直肠癌细胞生长和凋亡。     

微小RNA-140-3p靶向细胞分裂周期相关蛋白8抑制肺腺癌细胞侵袭转移的生物信息学分析及验证

目的 在生物信息学基础上探讨微小RNA（miR)-140-3p靶向细胞分裂周期相关蛋白8（CDCA8）抑制肺腺癌细胞的侵袭和转移。 方法 通过GEO数据库中的GEO2R分析肺腺癌芯片数据中差异表达的miRNA。TargetScanHuman7.2和miRWalk数据库查找miR-140-3p的靶基因。Cytoscape3.7.2软件筛选出Hub基因。GEPIA数据库查询靶基因在肺腺癌组织和正常肺组织中的表达水平,在肺腺癌不同分期中的表达水平以及靶基因的表达水平与肺腺癌患者总体生存率的关系。StarBase数据库查找miR-140-3p在肺腺癌中的生存分析及与CDCA8的表达水平在肺腺癌中的相关性。Real-time PCR检测miR-140-3p在正常肺上皮细胞BEAS-2B和肺腺癌细胞A549的表达水平以及感染效率。Real-time PCR和Western blotting实验检测过表达miR-140-3p后CDCA8 mRNA和蛋白的表达水平。双荧光素酶报告基因实验验证miR-140-3p是否与CDCA8直接结合。Transwell侵袭实验检测过表达miR-140-3p和CDCA8对肺腺癌细胞侵袭力的影响。 结果 通过GEO等数据库的分析结果显示,miR-140-3p在正常肺组织中的表达水平明显高于在肺腺癌中的表达水平,其预测靶基因CDCA8在肺腺癌中的表达水平明显高于在正常肺组织中的表达水平,且CDCA8与miR-140-3p的表达水平在肺腺癌中成负相关。实验结果显示,miR-140-3p在A549细胞中的表达明显低于在BEAS-2B细胞中的表达 (P＜0.05);用过表达慢病毒感染细胞后miR-140-3p的表达水平明显升高 (P＜0.05);过表达miR-140-3p后CDCA8的mRNA和蛋白表达水平明显下调 (P＜0.05);双荧光素酶报告基因实验结果显示,miR-140-3p能与CDCA8直接结合 (P＜0.05);与对照组相比,过表达miR-140-3p可抑制肺腺癌细胞A549的侵袭转移,而CDCA8可逆转miR-140-3p对肺腺癌细胞A549侵袭力的抑制作用 (P＜0.05)。 结论 MiR-140-3p靶向CDCA8抑制肺腺癌细胞的侵袭转移。     

Overexpression of integrin-linked kinase (ILK) promotes migration and invasion of colorectal cancer cells by inducing epithelial–mesenchymal transition via NF-κB signaling

Integrin-linked kinase (ILK), a ubiquitously expressed and evolutionally conserved serine/threonine kinase, has been shown to be aberrantly overexpressed and activated in diversified types of human malignancies, including colorectal cancer (CRC). However, the potential role of ILK in cancer cell migration and invasion remains to be elucidated. In this study, we introduced the human ILK gene into a low ILK-expressing human CRC cell line SW480. Cell migration and invasion were evaluated by the wound healing assay and transwell invasion assay, respectively. The epithelial–mesenchymal transition (EMT)-related proteins were detected by Western blot analysis or immunofluorescence. We found that enforced overexpression of ILK in SW480 cells dramatically promoted their migratory and invasive ability in vitro. Furthermore, SW480 cells stably overexpressing ILK underwent EMT, as indicated by mesenchymal morphology, decreased expression of E-cadherin, and increased expression of vimentin, Snail, and Slug. Finally, the nuclear factor (NF)-κB inhibitor BAY 11-7028 or NF-κB p65 small interfering RNA significantly restored the reduced E-cadherin level in ILK-overexpressing cells, suggesting that ILK-mediated down-regulation of E-cadherin is dependent on NF-κB activation. Overall, our study demonstrates a pivotal role of ILK in EMT and metastasis, and suggests novel therapeutic opportunities for the treatment of CRC.     

BDNF-AS/miR-145-5p轴对高糖诱导的肾小管上皮细胞损伤的影响CSCD

目的探讨脑源性神经营养因子反义RNA(brain-derived neurotrophic factor-antisense, BDNF-AS)对高糖诱导的肾小管上皮细胞损伤的影响和可能机制。方法体外培养肾小管上皮细胞HK-2, 分别转染BDNF-AS小干扰RNA、miR-145-5p模拟物或共转染BDNF-AS小干扰RNA和miR-145-5p抑制剂, 之后采用30 mmol/L葡萄糖干预转染后的细胞24 h, 用RT-qPCR法检测细胞中BDNF-AS和miR-145-5p的表达, 用CCK-8法检测细胞增殖, 流式细胞术检测细胞凋亡, Western印迹法检测细胞中Bcl-2和Bax蛋白的表达, 酶联免疫吸附法检测细胞培养上清中IL-1β和IL-6的水平。用双荧光素酶报告基因实验验证BDNF-AS和miR-145-5p的调控关系。结果高糖处理促进了HK-2细胞中BDNF-AS的表达(P<0.05), 而抑制了miR-145-5p的表达(P<0.05)。干扰BDNF-AS或过表达miR-145-5p降低了高糖诱导的HK-2细胞抑制率、凋亡率及Bax蛋白、IL-1β和IL-6的表达(P<0.05), 促进了Bcl-2蛋白的表达(P<0.05)。干扰miR-145-5p逆转了干扰BDNF-AS对高糖诱导的HK-2细胞增殖、凋亡及IL-1β和IL-6表达的影响。BDNF-AS可靶向结合并负调控miR-145-5p。结论干扰BDNF-AS可能通过靶向负调控miR-145-5p促进高糖诱导的肾小管上皮细胞增殖, 并抑制细胞凋亡及炎症因子表达。     

MicroRNA-98通过下调EZH2表达抑制结直肠癌细胞的活力与侵袭能力

目的:探讨微小RNA-98(mi R-98)与Zeste基因增强子同源物2(EZH2)之间的靶向关系,及其对结直肠癌细胞活力和侵袭能力的影响。方法:Target Scan软件预测EZH2和mi R-98之间的靶向结合,双萤光素酶报告基因实验验证mi R-98与EZH2之间的靶向关系。用mi R-98 mimic和mi R-98 inhibitor分别转染人结直肠癌SW480细胞和SW620细胞,Western blot检测EZH2的蛋白表达; MMT法检测细胞活力; Transwell法检测细胞的侵袭能力。转染EZH2过表达质粒检测其对细胞活力和侵袭的影响。结果:mi R-98能够靶向调节EZH2并负向调节SW480细胞和SW620细胞中EZH2的蛋白表达。在SW480细胞和SW620细胞中,过表达mi R-98显著下调细胞活力和侵袭能力,而抑制mi R-98表达显著促进细胞生长和侵袭。过表达EZH2也可促进SW480细胞和SW620细胞的生长和侵袭。结论:mi R-98通过下调EZH2表达抑制SW480细胞和SW620细胞的活力和侵袭。本研究为结直肠癌的治疗提供了新的靶点和方向。     

MiR-145-5p在大肠癌中的表达及与多药耐药的关系

目的 探讨大肠癌中miR-145-5p和多药耐药基因(MDR1)蛋白P-糖蛋白（P-gp）的表达及两者的关系。方法 分别在组织、细胞水平采用SP法、Real-time PCR法、Western blotting法检测50例大肠癌 (CRC)患者组织及30例癌旁正常组织患者中P-gp的表达,miR-145-5p的表达变化对MDR1 mRNA及P-gp的影响及两者与临床病理特征之间的相关性。结果 大肠癌组织中miR-145-5p表达量明显低于癌旁正常组织,MDR1 mRNA及P-gp的表达量明显高于癌旁正常组织(r=-0.403,P<0.01)。大肠癌细胞(HCT-15)中miR-145-5p 能够抑制MDR1 mRNA及P-gp的表达(P<0.05)。结论 MiR-145-5p对大肠癌多药耐药基因及蛋白的表达具有调控作用,参与了大肠癌的发生、发展及多药耐药。     

CircCBFB is a mediator of hepatocellular carcinoma cell autophagy and proliferation through miR-424-5p/ATG14 axis

This study aims to investigate the role of circCBFB in hepatocellular carcinoma (HCC) cell proliferation and autophagy. qRT-PCR and Western blotting analyses quantified the expression levels of circCBFB, miR-424-5p, and ATG14 in HCC tissues and/or HCC cell lines. After transfection with pcDNA3.1-CircCBFB, sh-CircCBFB, miR-424-5p mimic, miR-424-5p inhibitor, pcDNA3.1-ATG14, sh-ATG14, sh-CircCBFB?+?miR-424-5p inhibitor, pcDNA3.1-CircCBFB?+?miR-424-5p mimic, sh-CircCBFB?+?pcDNA3.1-ATG14, or pcDNA3.1-CircCBFB?+?sh-ATG14, the proliferation, cell cycle, and apoptosis of Huh-7 and HCCLM3 cells were detected, respectively, through MTT assay and flow cytometry. Western blotting measured the expression levels of ATG14 and autophagy-related proteins (LC3-ΙΙ/LC3-Ι, Beclin1, and p62). The interactions among circCBFB, miR-424-5p, and ATG14 were identified through RNA fluorescence in situ hybridization and RNA immunoprecipitation. In HCC tissues, circCBFB and ATG14 were highly expressed, and miR-424-5p expression was downregulated. Transfection of pcDNA3.1-CircCBFB, miR-424-5p inhibitor, or pcDNA3.1-ATG14 into HCC cells facilitated HCC cell proliferation and autophagy, while suppressing cell apoptosis, evidenced by elevated cell viability, increased protein levels of autophagosome markers (LC3-ΙΙ/LC3-Ι and Beclin1), repressed apoptosis rate, and suppressed protein level of autophagy receptor p62. miR-424-5p was a target gene of circCBFB, and miR-424-5p negatively mediated ATG14. CircCBFB inhibits miR-424-5p and upregulates ATG14, thus promoting HCC cell proliferation and autophagy.

     

Lidocaine inhibits proliferation and induces apoptosis in colorectal cancer cells by upregulating mir-520a-3p and targeting EGFR

Lidocaine is a conventional local anesthetic which is shown antiproliferative of colorectal cancer (CRC) in patients. MicroRNAs (miRNAs) have been consistently demonstrated to be involved in CRC, and miR-520a-3p could suppress CRC migration, promote apoptosis by targeting epidermal growth factor receptor (EGFR). However, the mechanism by which lidocaine regulated CRC proliferation and apoptosis remains unknown. In this study, quantitative RT-PCR were used to measure miR-520a-3p and EGFR expression levels, and western blotting assays ware performed to measure EGFR expression in CRC cells. Luciferase reporter assay was employed to validate the direct targeting of EGFR by miR-520a-3p. Cell proliferation and apoptosis assays ware utilized to analyze the role of lidocaine in CRC cells. The results indicated that 500 and 1000 μM lidocaine over 24?h inhibited proliferation and induced apoptosis of CRC cells. Compared with the control group, the expression of EGFR was suppressed by lidocaine (500 μM) in CRC cells. Furthermore, miR-520a-3p could directly targets EGFR in CRC cells. Lidocaine (500 μM) increased the expression of miR-520a-3p and rescued the reduction of miR-520a-3p caused by miR-520a-3p inhibitor. The results suggested that lidocaine could suppress the expression of EGFR by upregulating miR-520a-3p, and it could induce apoptosis and inhibit proliferation in CRC cells. Lidocaine may serve as potential therapeutic regimen for colorectal cancer.     

ADD3可变剪接在结肠癌与正常组织间的差异表达

 目的: 探讨内收蛋白3(ADD3)及其剪接体与结直肠癌(CRC)的关系。方法: Real-time PCR方法检测50对CRC组织、20对结直肠息肉组织、经5-氟尿嘧啶(5-FU)或奥沙利铂干预前后的SW480(低转移性)和SW620(高转移性)结肠癌细胞中ADD3-、ADD3-Ia和ADD3-Ib的表达量。用MTT检测细胞的活力,用划痕实验检测细胞的转移能力,用Transwell实验检测细胞的侵袭性。结果: CRC组织中ADD3与ADD3-Ib的表达量低于正常组织(P<0.01),ADD3-Ia/Ib比值高于正常组织(P<0.01);病理分组发现ADD3-Ia在T3-4组的表达量较T1-2组高(P<0.05)。结直肠息肉组织中ADD3、ADD3-Ia和ADD3-Ib的表达量低于正常组织(P<0.01)。SW620细胞中ADD3-Ia和ADD3-Ib的表达量低于SW480细胞(P<0.05),ADD3-Ia/Ib比值高于SW480(P<0.01);在经奥沙利铂和5-FU分别处理后,在SW620和SW480细胞活力、迁移和侵袭能力减弱,而ADD3、ADD3-Ia和ADD3-Ib的表达量均有不同程度增高。结论: CRC中ADD3-Ib和ADD3减少,并伴随ADD3-Ia/Ib比值升高。ADD3及其剪接体的改变可能跟CRC的侵袭能力有关。     

miR-197-3p调控DMBT1抑制甲状腺癌细胞系增殖、迁移和侵袭

目的探讨miR-197-3p是否通过靶向调控恶性脑瘤缺失1基因(DMBT1)影响甲状腺癌细胞增殖、迁移和侵袭。方法RT-qPCR检测健康人甲状腺细胞Nthy-ori 3-1和甲状腺癌细胞SW579、CGTHW-1中miR-197-3p表达;MTT法检测SW579细胞增殖;Transwell小室法检测SW579细胞迁移和侵袭;双荧光素酶报告基因实验验证miR-197-3p是否靶向DMBT1;Western blot检测细胞DMBT1、cyclinD1、p21、MMP-2和E-cadherin蛋白表达。结果与Nthy-ori 3-1细胞比较,SW579和CGTHW-1细胞中miR-197-3p相对表达量升高(P<0.05);抑制miR-197-3p表达后,SW579细胞的增殖、迁移和侵袭能力明显受到抑制,细胞中cyclinD1蛋白和MMP-2蛋白表达降低而p21蛋白和E-cadherin蛋白表达升高;SW579细胞中miR-197-3p靶向负调控DMBT1的表达;过表达DMBT1明显抑制SW579细胞增殖、迁移和侵袭,而抑制DMBT1则能逆转miR-197-3p对SW579细胞增殖、迁移和侵袭的影响。结论miR-197-3p通过靶向调控DMBT1的表达,抑制甲状腺癌细胞增殖、迁移和侵袭。     

miR-125b通过下调HAX-1的表达提高CD133~+ SW480细胞对顺铂的敏感性

目的:探讨miR-125b在CD133~+结直肠癌细胞中发挥的作用并研究其是否和顺铂的体外治疗有关。方法:用RT-qPCR方法检测miR-125b在常规SW480肿瘤细胞及CD133~+ SW480肿瘤细胞中的表达水平。流式细胞术检测miR-125b和顺铂对SW480细胞系中CD133~+ 细胞比例的影响。MTT法检测miR-125b对顺铂杀伤CD133~+ SW480细胞能力的影响。利用生物信息学及Western blot方法验证miR-125b是否调节CD133~+ SW480细胞中HAX-1的表达。运用JC-1染色、Annexin V染色及Western blot方法研究miR-125b影响顺铂疗效的信号通路的效应。结果:CD133~+ SW480细胞中的miR-125b表达水平显著低于正常结直肠上皮细胞系FHC和常规SW480肿瘤细胞。顺铂体外单独治疗能提高SW480细胞系中CD133~+细胞的比例,然而联用miR-125b模拟物后CD133~+ SW480细胞的比例显著下降。MTT实验结果表明miR-125b可显著增强顺铂对CD133~+ SW480细胞的杀伤活性。Western blot实验表明miR-125b的靶基因可能为HAX-1。miR-125b联合顺铂可引起CD133~+ SW480细胞线粒体膜电位的丧失并诱导线粒体内细胞色素C的释放,进而引起细胞凋亡。转染HAX-1表达载体后miR-125b联合顺铂对CD133~+ SW480细胞的凋亡诱导效应显著降低。结论:miR-125b通过下调HAX-1的表达提高CD133~+结直肠癌细胞对顺铂的敏感性。     

miR-145-5p靶向TPM3通过ERK信号通路抑制甲状腺癌TPC-1细胞侵袭和转移

目的 探讨miR-145-5p对甲状腺癌细胞侵袭和转移的影响.方法 将miR-145-5p mimic质粒、mimic-NC质粒、TPM3质粒分别或联合转入甲状腺癌细胞,RT-qPCR检测miR-145-5p与TPM3 mRNA的表达,双荧光素酶报告检测靶向关系,Transwell法检测细胞侵袭能力,划痕实验检测细胞迁...     

MicroRNA143调节结肠癌细胞KRAS蛋白的表达

目的：探讨结肠癌细胞中microRNA143（miR-143）对KRAS蛋白表达的影响及其作用机制。方法: 从基因组DNA中克隆pri-miR-143基因，构建miR-143的表达载体。miR-143转染结肠癌细胞系SW480，用Northern blotting方法测定miR-143水平的变化，通过RT-PCR和Western blotting方法检测转染细胞中KRAS基因的表达。构建含miR-143结合位点的荧光素酶报告载体，与miR-143表达载体共转染，检测细胞中的荧光素酶活性。结果: 基因转染的SW480细胞中miR-143的表达显著升高， KRAS蛋白的表达水平下降约40%，KRAS mRNA则未受影响。miR-143表达载体与含miR-143结合位点的报告载体共转染可以抑制细胞中的荧光素酶活性。结论: 结肠癌细胞中microRNA143在转录后水平抑制KRAS蛋白的表达。     

miR-490-3p通过抑制TGFβR1基因表达进而抑制结肠癌细胞的转移和侵袭

目的：研究miR-490-3p在结肠癌细胞(colorectal cancer cell,CRC)转移中的表现和生物功能,及其调控作用机制。方法：通过荧光定量PCR测定miR-490-3p在CRC细胞系的表达水平。细胞转染miR-490-3p以及shmiR-490-3p,观察miR-490-3p的过表达或基因沉默对结肠癌细胞的转移能力是否有影响。miR-490-3p的分子靶标由双荧光素酶报告基因分析和免疫印迹技术进行实验认定。通过划痕实验,Transwell小室基质渗透实验对细胞迁移和侵袭能力进行鉴定。结果：miR-490-3p在CRC细胞系中显著低表达(P<0.05,P<0.01,P<0.001,n>3)。过表达miR-490-3p显著降低CRC细胞株的细胞迁移和侵袭能力(P<0.01,n>3)。miR-490-3p的基因沉默显著增加CRC细胞株的细胞迁移和侵袭能力(P<0.01,n>3)。结肠癌细胞细胞系中过表达miR-490-3p显著降低TGFβR1的基因表达(P<0.001,n>3),miR-490-3p基因沉默显著上调TGFβR1的基因表达(P<0.001,n>3)。过表达miR-490-3p抑制TGFβR1的萤光素酶活性(P<0.001,n>3),miR-490-3p基因沉默促进TGFβR1的萤光素酶活性(P<0.001,n>3)。TGFβR1基因沉默减弱shmiR-490介导的细胞迁移和侵袭促进效应(P<0.01, n>3)。结论：miR-490-3p通过抑制TGFβR1的基因表达从而抑制CRC细胞的转移。     

MiR-876-5p acts as an inhibitor in hepatocellular carcinoma progression by targeting DNMT3A

Hepatocellular carcinoma (HCC) is one of the biggest challenges that human beings faced with in 21st century. Previous researches have revealed that miRNAs can serve as regulators in various cancers. MiR-876-5p, a member of miRNA family, has been studied in lung cancer for its anti-oncogenic function. However, the exact function of it is not reported in HCC. Our study aims to find out the effects of miR-876-5p expression on HCC progression. Two HCC cells were chosen to do functional assays after miR-876-5p expression was detected in cell lines by qRT-PCR. HepG2 cell was transfected with miR-876-5p mimics, whereas LM3 cell was transfected with miR-876-5p inhibitors. Next, cell activities of these two indicated cells were analyzed by means of MTT assay, colony forming assay, transwell migration assay and western blot analysis. Consequently, we found that miR-876-5p could inhibit both cell proliferation and metastasis. Moreover, we found out a target gene (DNMT3A) of miR-876-5p by performing bioinformatics analysis, dual luciferase reporter assay and biotin-avidin pull-down assay. Finally, rescue assays were carried out in HepG2 cells. We found that DNMT3A could reverse miR-876-5p mimics-induced inhibition. Therefore, we concluded that miR-876-5p suppressed hepatocellular carcinoma progression by targeting DNMT3A.