miR-125a-5p通过靶向调控GAB2抑制乳腺癌细胞的迁移能力

miR-125a-5p可负性调节GAB2表达,抑制胶质瘤细胞的侵袭和转移。本研究旨在证明miR-125a-5p抑癌作用的普遍性,即miR-125a-5p是否可通过靶向抑制GAB2抑制乳腺癌细胞的迁移。荧光素酶实验结果显示,miR-125a-5p可特异识别GAB2的3′-UTR,抑制报告酶的表达。荧光定量PCR结果揭示,与正常乳腺上皮细胞MCF-10A比较,miR-125a-5p在乳腺癌细胞MDA231和MCF-7中的表达明显降低;与迁移能力相对较低的MCF-7细胞比较,miR-125a-5p在迁移能力较高的MDA231细胞中的表达量更低。Western 印迹结果证明,与空载体（对照）和anti-miR125a 5p转染细胞比较,转染miR-125a-5p明显抑制GAB2蛋白在乳腺癌细胞中的表达。Transwell结果显示,与空载体转染的对照细胞比较,转染miR-125a-5p的乳腺癌细胞穿过基质胶的细胞数明显减少;相反,转染anti-miR125a-5p的细胞穿过基质胶的细胞数却明显增多。上述结果提示,miR-125a-5p在正常的乳腺细胞中高表达,而在乳腺癌细胞中低表达,其表达水平与癌细胞的迁移能力和GAB2表达呈反向关系。本研究结果还提示,miR-125a-5p通过靶向负调控GAB2抑制乳腺癌细胞的迁移能力。总之,本研究证明,miR-125a-5p在肿瘤中发挥抑癌作用。     

circPVT1通过miR-24-3p/let-7a-5p/FSCN1轴促进鼻咽癌细胞侵袭迁移

目的 鼻咽癌是一种来源于鼻咽上皮的恶性肿瘤,其临床特征之一是易发生淋巴转移,但是目前鼻咽癌转移的分子机制尚未阐明。circPVT1是由PVT1基因2号外显子反向拼接形成的环状RNA (circRNA),在多种肿瘤中表达上调,本文探讨了circPVT1在鼻咽癌侵袭迁移中的作用和分子机制。方法 通过RT-qPCR检测circPVT1及其下游miRNA和FSCN1在鼻咽癌细胞的表达情况,Transwell和划痕愈合实验检测circPVT1对鼻咽癌细胞侵袭迁移的影响,RNA pull-down实验检测circPVT1结合的miRNA,双荧光素酶报告实验检测miR-24-3p和let-7a-5p靶向抑制FSCN1 mRNA表达。结果 在鼻咽癌细胞中过表达circPVT1可以促进鼻咽癌细胞侵袭迁移,而敲低circPVT1则可以抑制鼻咽癌细胞的侵袭迁移。进一步研究发现,circPVT1可以通过竞争性吸附miR-24-3p和let-7a-5p,上调FSCN1的表达,从而促进鼻咽癌细胞的侵袭迁移。结论 circPVT1通过miR-24-3p/let-7a-5p/FSCN1轴促进鼻咽癌细胞侵袭迁移,证实c...     

环状RNA circMRPS35通过miR-130a-3p/ZNRF3轴调节胃癌细胞的增殖、凋亡、迁移和侵袭

摘要 目的：探讨环状RNA MRPS35（circMRPS35）对胃癌（GC）细胞增殖、凋亡、迁移和侵袭的调控机制。方法：体外培养人GC细胞系（HGC-27、MGC-803、MKN45和AGS）和正常胃上皮GES-1细胞,实时荧光定量PCR（RT-qPCR）检测circMRPS35、miR-130a-3p和锌环指蛋白3（ZNRF3）mRNA表达。另取MGC-803细胞,分为对照组、pc-NC组、pc-circMRPS35组、pc-circMRPS35+miR-NC组、pc-circMRPS35+miR-130a-3p组,采用Lipofectamine 3000进行质粒转染。RT-qPCR检测circMRPS35、miR-130a-3p和ZNRF3 mRNA表达,Western blot检测ZNRF3蛋白表达,CCK-8法、流式细胞术检测细胞增殖与凋亡,划痕实验和Transwell小室实验检测细胞迁移与侵袭能力,裸鼠移植瘤实验探究circMRPS35对GC细胞体内生长的影响。双荧光素酶报告基因检测miR-130a-3p与circMRPS35或ZNRF3的靶标关系。结果：GC细胞系中circMRPS35和ZNRF3 mRNA呈低表达,miR-130a-3p呈高表达（均P＜0.05）。过表达circMRPS35可降低miR-130a-3p,上调ZNRF3 mRNA和蛋白水平,抑制细胞增殖、迁移和侵袭,并促进细胞凋亡（均P＜0.05）;circMRPS35过表达对GC细胞恶性行为和裸鼠移植瘤生长的抑制作用可被miR-130a-3p mimic逆转（P＜0.05）。双荧光素酶实验结果显示,过表达miR-130a-3p可降低circMRPS35-WT和ZNRF3-WT的荧光素酶活性（P＜0.05）。结论：circMRPS35可能通过miR-130a-3p/ZNRF3轴抑制GC细胞的增殖、迁移和侵袭,并促进细胞凋亡。     

miR-19a-3p在前列腺癌细胞侵袭和迁移中的作用

miR-19a-3p通过多种机制调节癌细胞的增殖和转移,然而,miR-19a-3p在前列腺癌转移中的生物学作用和机制尚不清楚。本研究旨在考察mir-19a-3p在前列腺癌中的表达情况,及其对细胞侵袭和迁移的影响。本研究发现,miR-19a-3p在骨转移性前列腺癌组织和前列腺癌细胞中明显下调。上调miR-19a-3p可显著抑制前列腺癌细胞的侵袭和迁移。然而,下调miR-19a-3p则会逆转上述变化。此外,本研究还发现miR-19a-3p通过靶向TGF-β信号传导的下游效应物SMAD2来抑制前列腺癌细胞中TGF-β信号传导的活性,从而抑制前列腺癌细胞的侵袭和迁移。因此,本研究表明mir-19a-3p与前列腺癌的发生发展密切相关,mir-19a-3p有望成为前列腺癌的新治疗靶点及生物标志物。     

骨髓间充质干细胞外泌体miR-190a-5p通过靶向KLF15抑制肺癌细胞迁移和侵袭

目的:探讨骨髓间充质干细胞(BMSC)来源的外泌体miR-190a-5p对肺癌细胞的影响。方法:通过超速离心获得BMSCs外泌体,透射电镜观察外泌体形态,采用纳米颗粒示踪分析(NTA)检测外泌体粒径,利用Western印迹检测外泌体上的标志蛋白CD63、CD9及HSP70;选取肺癌细胞系A549、LK79、H1975和HCC827,以及人正常上皮细胞BEAS-2B检测对比miR-190a-5p在这些细胞中和BMSCs衍生的外泌体(BMSC-exosome)中的表达量;双萤光素酶报告基因检测验证Krüppel样因子15(KLF15)是否为miR-190a-5p的靶基因;定量PCR(qRT-PCR)和Western印迹检测miR-190a-5p对KLF15的表达调控;Transwell法检测外泌体对肺癌细胞迁移和侵袭的影响。结果:BMSCs外泌体呈圆形,粒径集中在150～200 nm,标志蛋白CD63、CD9及HSP70阳性表达;BMSCs外泌体中miR-190a-5p的相对表达量均高于在4种肺癌细胞及正常肺细胞BEAS-2B中的表达;双萤光素酶报告基因检测KLF15是miR-190a-5p的靶基因;BMSCs外泌体与miR-190a-5p mimics均能使肺癌细胞中的miR-190a-5p含量升高,并抑制KLF15的mRNA和蛋白表达,从而抑制肺癌细胞迁移和侵袭。结论:BMSCs外泌体miR-190a-5p通过下调KLF15抑制肺癌细胞迁移和侵袭,为肺癌的诊断和治疗提供了新的思路。     

miR-193a-5p促进人胰腺癌细胞体外迁移和侵袭作用的研究

该研究主要探讨了微小核酸mi RNA-193a-5p对人胰腺癌细胞体外迁移和侵袭的促进作用及其机制。采用miR-193a-5p模拟物及mi R-193a-5p抑制剂分别上调和下调miR-193a-5p的表达;采用细胞划痕法和Transwell小室法检测mi R-193a-5p对细胞迁移和侵袭能力的影响;采用TargetScan 7.1数据库预测miR-193a-5p的靶基因;荧光素酶报告法验证miR-193a-5p的靶基因; Western blot和实时荧光定量PCR验证其表达结果。结果表明, miR-193a-5p可显著促进细胞的迁移和侵袭能力。TargetScan 7.1软件预测, Prox1可能为mi R-193a-5p的靶基因,荧光素酶报告实验显示, miR-193a-5p靶向Prox1基因的3′UTR区。实时荧光定量PCR和Western blot结果显示, miR-193a-5p下调了Prox1的mRNA和蛋白水平的表达。研究结果揭示,在胰腺癌中高表达的miR-193a-5p通过下调Prox1,从而使胰腺癌细胞获得高的迁移和侵袭能力,促进胰腺癌的转移。     

MiR-30a-3p 对人滋养肿瘤细胞系JEG-3 细胞侵袭功能的影响

目的：MiRNAs 对于胎盘的形成和正常妊娠的维持起着至关重要的作用,它在胎盘中的表达失衡的可能导致了妊娠相关疾 病的发生,我们前期研究发现miR-30a-3p 在子痫前期患者胎盘上特异性高表达,推测miR-30a-3p 可能参与了子痫前期的发生发 展过程,本课题通过观察miR-30a-3p 对人滋养肿瘤细胞系JEG-3 细胞侵袭能力的影响,深入探讨miR-30a-3p 在子痫前期发病过 程中的作用。方法：应用瞬时转染技术在人滋养肿瘤细胞系JEG-3 细胞中分别转染miR-30a-3p mimics、mimics NC为miR-30a-3p 过表达组和阴性对照组,空白转染组为空白对照组,利用荧光实时定量PCR 技术检测各组细胞中miR-30a-3p的表达,Transwell 实验检测各组细胞侵袭能力的差别。结果：荧光实时定量PCR结果显示miR-30a-3p 过表达组与阴性对照组、空白对照组相比 miR-30a-3p 的表达量明显升高,差异具有统计学意义（P<0.05）;Transwell 实验结果显示miR-30a-3p过表达组细胞的侵袭能力与 阴性对照组、空白对照组相比均有降低,差异具有统计学意义(P<0.05)。阴性对照组与空白对照组的侵袭能力差异无统计学意义 (P>0.05)。结论：miR-30a-3p 可以显著下调JEG-3 细胞的侵袭力, miR-30a-3p 有可能通过降低滋养细胞的浸润能力,导致滋养细胞 对子宫肌层和螺旋动脉的浸润不足,造成“胎盘浅着床”,从而在子痫前期的发病过程中发挥了重要的作用,miR-30a-3p 有望成为 诊治子痫前期疾病的靶点。     

miR-143-3p通过靶向TAK1抑制肺癌增殖和侵袭

目的:分析miR-143-3p在肺癌细胞中的表达情况以及对肺癌进展的作用。方法:通过starBase数据库和肺癌病例组织检测分析miR-143-3p在肺癌组织和正常对照组织中的表达差异;分析miR-143-3p在肺癌细胞HCC27、H1975、A549和肺上皮细胞BEAS-2B中的mRNA水平差异;采用CCK-8法检测miR-143-3p对肺癌细胞增殖活性的影响;采用Transwell实验检测miR-143-3p对肺癌细胞迁移和侵袭的影响;通过qRT-PCR检测miR-143-3p对整合素α6(ITGA6)、锚蛋白重复及PH结构域3(ASAP3)、黑色素瘤相关抗原A9(MAGE-A9)和转化生长因子β激活激酶1(TAK1)表达的影响;通过Western印迹检测miR-143-3p对TAK1蛋白表达的影响。结果:starBase分析和肺癌病例组织检测结果显示miR-143-3p在肺癌组织中低表达,同样地,miR-143-3p在肺癌细胞中的表达也显著低于正常肺上皮细胞;过表达miR-143-3p抑制了肺癌细胞的增殖活性、迁移和侵袭能力;过表达miR-143-3p显著抑制TAK1的表达。结论:miR-143-3p在肺癌中通过靶向TAK1抑制肺癌的增殖和侵袭,miR-143-3p在肺癌进展中详细的分子作用机制和信号通路仍须进一步探讨。     

DLC-1基因表达对结肠癌细胞侵袭转移能力的影响

目的:研究DLC-1基因对结肠癌细胞侵袭迁移能力的影响.方法:将DLC-1 shRNA(短发夹状RNA,short hairpin RNA)序列克隆到质粒pGCsi-U6/Neo载体,采用脂质体介导的转染方法将构建的DLC-1 shRNA表达质粒转入结肠癌细胞系LoVo细胞.采用RT-PCR技术和Western Blot技术分别检测LoVo细胞中DLC-1mRNA和蛋白表达水平的变化.Transwell小室人工重组基底膜侵袭转移实验观察LoVo细胞侵袭迁移能力的改变.结果:结肠癌细胞系LoVo细胞表达DLC-1分子.所构建质粒表达载体能有效地干扰LoVo细胞DLC-1 mRNA和蛋白质表达水平;Transwell小室人工重组基底膜侵袭转移实验结果显示,转染后LoVo细胞侵袭转移能力明显增强(p＜0.05).结论:结肠癌细胞系LoVo细胞表达DLC-1基因,应用RNAi技术可特异性降低其表达.DLC-1的表达水平与结肠癌细胞侵袭转移相关.     

环状RNA circTCF25通过miR-103a-3p/miR-107调控CDK6的表达促进膀胱癌的增殖和迁移

该文旨在探讨环状RNA circTCF25对膀胱癌增殖和迁移的影响。采用RT-q PCR检测膀胱癌及癌旁组织中miR-103a-3p和miR-107的表达水平;采用Western blot和免疫组化检测膀胱癌细胞及膀胱癌和癌旁组织中CDK6的蛋白表达;将circTCF25过表达载体转染两种膀胱癌细胞后,采用RT-q PCR检测细胞中circTCF25以及miR-103a-3p和miR-107的表达水平;采用双荧光素酶报告基因实验验证circTCF25靶向结合miR-107及miR-107的靶基因CDK6;划痕实验检测细胞迁移能力;Edu实验检测细胞增殖能力。RT-q PCR结果表明,膀胱癌组织中miR-103a-3p和miR-107的表达明显低于癌旁组织。免疫组化和Western blot结果显示,膀胱癌组织中CDK6蛋白质水平明显高于癌旁组织。转染circTCF25过表达载体的细胞中,circTCF25的表达水平高于对照组。过表达circTCF25后,细胞中miR-103a-3p和miR-107的表达显著下降,CDK6的蛋白水平增加。双荧光素酶报告基因实验表明,circTCF25可以直接结合miR-107并降低其对靶基因CDK6的抑制。过表达circTCF25后,细胞迁移和增殖能力增强。该研究说明,环状RNA circTCF25可通过miR-103a-3p/miR-107调控CDK6的表达促进膀胱癌的增殖和迁移。     

miR-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为研究

目的:探讨mi R-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为。方法:qRT-PCR法检测并比较肺癌组织、癌旁正常组织、肺癌细胞、正常肺上皮细胞中的mi R-199a-3p m RNA相对表达量。比较远处转移肺癌组织、未转移肺癌组织中mi R-199a-3p m RNA相对表达量。qRT-PCR法、Western Blot法检测并比较肺癌组织、癌旁正常组织中的CBX7 m RNA及蛋白的表达水平。荧光素酶活性法检测mi R-199a-3p与靶基因CBX7的结合。比较mi R-199a-3p模拟物转染组与阴性对照组的肺癌细胞中的CBX7 m RNA相对表达量及CBX7蛋白表达水平。CCK8实验检测mi R-199a-3p对肺癌细胞增殖的促进作用。Tranwell实验检测mi R-199a-3p对肺癌细胞侵袭与迁移能力的影响。结果:肺癌组织中mi R-199a-3p明显高于癌旁正常组织,发生远处转移的肺癌组织中mi R-199a-3p m RNA的表达量明显高于未发生转移的肺癌组织,差异有统计学意义(P<0.001)。肺癌组织中CBX7m RNA、CBX7蛋白表达水平均明显低于癌旁正常组织,差异有统计学意义(P<0.001)。荧光素酶活性法证实mi R-199a-3p可与靶基因CBX7结合抑制CBX7的表达。肺癌细胞中mi R-199a-3p m RNA的相对表达量明显高于正常肺上皮细胞,CBX7 m RNA相对表达量明显低于正常肺上皮细胞(P<0.05)。对于肺癌细胞,mi R-199a-3p模拟物转染组的CBX7 m RNA相对表达量及CBX7蛋白表达水平均明显低于阴性对照组(P<0.001)。CCK8实验证实mi R-199a-3p能够促进肺癌细胞的增殖,Tranwell实验证实mi R-199a-3p对肺癌细胞侵袭与迁移具有积极的促进作用。结论:mi R-199a-3p在肺癌的发生发展过程中发挥重要作用,能够通过抑制CBX7基因的表达,促进肺癌细胞的增殖、侵袭和转移。     

MiR-216a-3p regulates the proliferation,apoptosis, migration,and invasion of lung cancer cells via targeting COPB2

ABSTRACT

Effect of miR-216a-3p on lung cancer hasn’t been investigated. Here, we explored its effects on lung cancer. MiR-216a-3p expression in lung cancer tissues and cells was detected by RT-qPCR. The target gene of miR-216a-3p was predicted by bioinformatics and confirmed by luciferase-reporter assay. After transfection, cell viability, migration, invasion, proliferation, and apoptosis were detected by MTT, scratch, transwell, colony formation, and flow cytometry. The expressions of COPB2 and apoptosis-related factors were detected by RT-qPCR or western blot. MiR-216a-3p was low-expressed and COPB2 was high-expressed in lung cancer tissues and cells. MiR-216a-3p targeted COPB2 and regulated its expression. MiR-216a-3p inhibited lung cancer cell viability, migration, invasion, and proliferation, while promoted apoptosis. Effect of miR-216a-3p on lung cancer was reversed by COPB2. MiR-216a-3p regulated proliferation, apoptosis, migration, and invasion of lung cancer cells via targeting COPB2.     

LncRNA-H19 activates CDC42/PAK1 pathway to promote cell proliferation,migration and invasion by targeting miR-15b in hepatocellular carcinoma

BackgroundHepatocellular carcinoma (HCC) is one of the main causes of cancer-related death. This study aims to explore the role and underlying mechanism of H19 in HCC.MethodsqRT-PCR detected miR-15b-5p and H19 expression, as well as the mRNA level of EMT-associated genes. Western blotting detected protein level of EMT-associated genes. Immunohistochemistry (IHC) examined CDC42 in HCC tissues. Dual luciferase reporter assay verified the regulatory mechanism among H19, miR-15b and CDC42. Colony formation, wound healing assay, transwell, flow cytometry measured proliferation, migration, invasion and apoptosis, respectively.ResultsH19 and CDC42 were up-regulated while miR-15b was down-regulated in HCC cells and tissues. miR-15b interacted with H19 and CDC42 3′-UTR. H19 knockdown inhibited proliferation, migration and invasion, and increased apoptosis, which was rescued by miR-15b inhibitor. H19 knockdown suppressed CDC42/PAK1 pathway and EMT progress.ConclusionH19 knockdown inhibited proliferation, migration and invasion, and promoted apoptosis of HCC cells via targeting miR-15b/CDC42/PAK1 axis.     

LncRNA SNHG12 promotes proliferation and epithelial mesenchymal transition in hepatocellular carcinoma through targeting HEG1 via miR-516a-5p

Hepatocellular carcinoma (HCC) is the most common cancer and its prognosis is poor due to metastasis and recurrence. EMT is associated with metastasis. A deep understanding of regulatory mechanism of EMT is critical. LncRNA is involved in regulation of various biological processes including EMT. This study aimed to investigate the regulatory signal axis among lncRNA SNHG12, miR-516a-5p and the target gene HEG1 during EMT. Cell cycle and apoptosis were analyzed by flow cytometry. Tumorigenesis was analyzed by clone formation assay. Wound healing assay and transwell assay was performed to detect migration and invasion, respectively. Interaction among SNHG12, miR-516a-5p and HEG1 were analyzed by dual luciferase assay and RIP assay. We also detected expression of RNA and protein by QPCR and western blotting. Finally, tumor growth was analyzed by tumorigenesis assay in vivo. Ki-67 and HEG1 level in tumor tissues was analyzed by IHC. SNHG12 and HEG1 were upregulated, miR-516a-5p was downregulated in HCC cell lines. SNHG12 could interact with and inhibit miR-516a-5p. MiR-516a-5p could interact with HEG1 and inhibit HEG1 expression. Knock down SNHG12 inhibited proliferation, migration, invasion, EMT and promoted apoptosis of HCC cells. Such effects were antagonized by inhibiting miR-516a-5p. SNHG12 overexpression lead to opposite results. Similar results were observed in mice. SNHG12 could promote EMT in HCC through targeting and inhibiting miR-516a-5p, which eventually upregulated HEG1 expression, in both cell and mice.     

MiR-517a-3p accelerates lung cancer cell proliferation and invasion through inhibiting FOXJ3 expression

Aims

Aberrant expression of microRNAs (miRNAs) results in alterations of various biological processes (e.g., cell cycle, cell differentiation, and apoptosis) and cell transformation. Altered miRNAs expression was associated with lung carcinogenesis and tumor progression. This study aimed to investigate the function and underlying molecular events of miR-517a-3p on regulation of lung cancer cell proliferation and invasion.

Main methods

Transfected miR-517a-3p mimics or inhibitors into 95D and 95C cells respectively, the effects of miR-517a-3p on lung cancer cell proliferation, migration, and invasion were detected. Bioinformatics software forecasted potential target genes of miR-517a-3p and dual luciferase reporter gene system and western blot verified whether miR-517a-3p regulates FOXJ3 expression directly.

Key findings

MiR-517a-3p was differentially expressed in lung cancer 95D and 95C cell lines that have different metastatic potential. Manipulation of miR-517a-3p expression changed lung cancer cell proliferation, migration and invasion capacity. MiR-517a-3p directly regulated FOXJ3 expression by binding to FOXJ3 promoter.

Significance

This study demonstrated that miR-517a-3p promoted lung cancer cell proliferation and invasion by targeting of FOXJ3 expression.     

In vitro and in vivo anti-metastatic effect of the alkaliod matrine from Sophora flavecens on hepatocellular carcinoma and its mechanisms

BackgroundsHepatocellular carcinoma (HCC) is one of the most prevalent and lethal cancer with high metastasis and recurrence rates. Hypoxia-induced miRNAs and HIF-1α are demonstrated to play essential roles in tumor metastasis. Matrine (C₁₅H₂₄N₂O), an alkaloid extracted from Sophora flavescens Aiton, has been used as adjuvant therapy for liver cancer in China. The anti-metastasis effects of matrine on HCC and the underlying mechanisms remain poorly understood.PurposeWe aimed to investigate the effects of matrine on metastasis of HCC both in vitro and in vivo, and explored whether miR-199a-5p and HIF-1α are involved in the action of matrine.MethodsMTT method, colony formation, wound healing and matrigel transwell assays were performed to evaluate the effects of matrine on cell proliferation, migration and invasion. Nude mice xenograft model and immunohistochemistry (IHC) assay were employed to investigate the anti-metastatic action of matrine in vivo. Quantitative real-time PCR, western blot and dual luciferase reporter assay were conducted to determine the underlying mechanisms of matrine.ResultsMatrine exerted stronger anti-proliferative action on Bel7402 and SMMC-7721 cells under hypoxia than that in normoxia. Both matrine and miR-199a-5p exhibited significant inhibitory effects on migration, invasion and EMT in Bel7402 and SMMC-7721 cells under hypoxia. Further study showed that miR-199a-5p was downregulated in HCC cell lines, and this microRNA was identified to directly target HIF-1α, resulting in decreased HIF-1α expression. Matrine induced miR-199a-5p expression, decreased HIF-1α expression and inhibited metastasis of Bel7402 and SMMC-7721 cells, while miR-199a-5p knockdown reversed the inhibitory effects of matrine on cell migration, invasion, EMT and HIF-1α expression. In vivo, matrine showed significant anti-metastatic activity in the nude mouse xenograft model. H&E and IHC analysis indicated that lung and liver metastasis nodules were reduced, and the protein expression of HIF-1α and Vimentin were significantly decreased by i.p injection of matrine.ConclusionsMatrine exhibits significant anti-metastatic effect on HCC, which is attributed to enhanced miR-199a-5p expression and subsequently impaired HIF-1α signaling and EMT. These findings suggest that miR-199a-5p is a potential therapeutic target of HCC, and matrine may represent a promising anti-metastatic medication for HCC therapy.     

细胞周期蛋白依赖性激酶在miR-193a5p调控卵巢癌细胞增殖及上皮细胞间充质转变中的作用*

目的: 探讨miR-193a-5p靶向CDK14并调控卵巢癌细胞OVAC的增殖和上皮间充质转变(EMT)的作用。方法: 通过TargetScanHuman分析miR-193a-5p与CDK14的匹配情况,通过荧光素酶报告系统检测miR-193a-5p靶向CDK14情况;在miR-193a-5p mimics过表达或者miR-193a-5p inhibitor基因沉默miR-193a-5p的情况下,采用免疫印迹检测CDK14,EMT相关蛋白质E-cadherin、vimentin、fibronectin和N-cadherin的表达量,采用CCK-8检测卵巢癌细胞OVAC增殖情况, MMT检测卵巢癌细胞OVAC的细胞活力。结果: miR-193a-5p靶向CDK14的3‘UTR;过表达miR-193a-5后, CDK14的表达下降,EMT相关蛋白质E-cadherin的表达上升,vimentin、fibronectin和N-cadherin的表达下降,卵巢癌细胞OVAC的增殖和细胞活力均增加;同时,基因沉默miR-193a-5p后, CDK14的表达上升,EMT相关蛋白质E-cadherin的表达下降,vimentin、fibronectin和N-cadherin的表达量上升,卵巢癌细胞OVAC的增殖和细胞活力均减少。结论: miR-193a-5p通过靶向CDK14的3‘UTR降低卵巢癌细胞OVAC的增殖、细胞活力和EMT。     

CircATRNL1 increases acid-sensing ion channel 1 to advance epithelial-mesenchymal transition in endometriosis by binding to microRNA-103a-3p

Circular RNA ATRNL1 (circATRNL1) has been implicated in epithelial-mesenchymal transition (EMT) during endometriosis. Given the existing literature and our predictions through starBase in this research, it was assumed that circATRNL1 might orchestrate the microRNA (miR)? 103a-3p/acid-sensing ion channel 1 (ASIC1) axis to control EMT in endometriosis. To verify our hypothesis, we detect circATRNL1, miR-103a-3p, and ASIC1 expression in endometrial cancer cells (HEC-B, AN3-CA, KLE, HEC1-A, and Ishikawa). Ishikawa cells with the highest circATRNL1 level were selected as subjects, where circATRNL1, miR-103a-3p, or ASIC1 expression was knocked down. Scratch and Transwell assays were applied to assess cell migration and invasion, and CCK-8 and colony formation assays to detect cell proliferation. Western blot was used to measure E-cadherin, N-cadherin, Vimentin, and Slug expression to evaluate the EMT state. Furthermore, the binding of miR-103a-3p to circATRNL1 or ASIC1 was validated by luciferase reporter assay. CircATRNL1 and ASIC1 were upregulated but miR-103a-3p was downregulated in endometrial cancer cells. Mechanistically, circATRNL1 bound to miR-103a-3p to upregulate a target gene of miR-103a-3p, ASIC1. CircATRNL1 silencing contributed to the decline of proliferation, invasion, migration, and EMT in Ishikawa cells, while miR-103a-3p inhibitor reversed those changes. In addition, the EMT process was aggravated when miR-103a-3p was inhibited and this process was suppressed by silencing ASIC1 in the presence of downregulated miR-101a-3p. Our study supported that circATRNL1 might be a novel therapeutic candidate target for endometriosis treatment and provided unique insights into the molecular basis concerning the pathogenesis of endometriosis.