miR-21通过靶向PDCD4调控三阴性乳腺癌细胞的迁移和侵袭

目的： 研究miR-21在三阴性乳腺癌细胞MDA-MB-231中的表达，以及其是否通过调控PDCD4影响MDA-MB-231细胞的迁移和侵袭。方法： 采用实时定量PCR（qPCR）法检测MDA-MB-231细胞和正常乳腺细胞MCF-10A中miR-21和PDCD4 mRNA的表达。将MDA-MB-231细胞随机分为5组：空白对照组，转染miR-21模拟物组，模拟物对照组，转染miR-21抑制物组和抑制物对照组。采用Western blot法检测MDA-MB-231细胞PDCD4蛋白的表达；采用荧光素酶报告基因试剂盒检测转染不同载体后荧光强度的变化来判断miR-21的靶标；采用Transwell实验检测各组细胞的迁移和侵袭数目。结果： miR-21和PDCD4 mRNA在MDAMB-231细胞中的表达水平分别明显高于和低于MCF-10A细胞（P均 < 0.01）。过表达或抑制miR-21可调节PDCD4的表达水平。荧光素酶报告基因试剂盒检测结果显示miR-21可直接靶向调控PDCD4的表达。Transwell实验结果表明过表达miR-21表达能增强MDA-MB-231细胞的迁移和侵袭能力。结论： 在MDA-MB-231细胞中，miR-21通过靶向调控PDCD4表达影响细胞的迁移和侵袭。miR-21可能成为抑制三阴性乳腺癌迁移和侵袭的靶点。     

miR-21通过靶向PDCD4调控三阴性乳腺癌细胞的迁移和侵袭

目的： 研究miR-21在三阴性乳腺癌细胞MDA-MB-231中的表达,以及其是否通过调控PDCD4影响MDA-MB-231细胞的迁移和侵袭。方法： 采用实时定量PCR（qPCR）法检测MDA-MB-231细胞和正常乳腺细胞MCF-10A中miR-21和PDCD4 mRNA的表达。将MDA-MB-231细胞随机分为5组：空白对照组,转染miR-21模拟物组,模拟物对照组,转染miR-21抑制物组和抑制物对照组。采用Western blot法检测MDA-MB-231细胞PDCD4蛋白的表达;采用荧光素酶报告基因试剂盒检测转染不同载体后荧光强度的变化来判断miR-21的靶标;采用Transwell实验检测各组细胞的迁移和侵袭数目。结果： miR-21和PDCD4 mRNA在MDAMB-231细胞中的表达水平分别明显高于和低于MCF-10A细胞（P均 < 0.01）。过表达或抑制miR-21可调节PDCD4的表达水平。荧光素酶报告基因试剂盒检测结果显示miR-21可直接靶向调控PDCD4的表达。Transwell实验结果表明过表达miR-21表达能增强MDA-MB-231细胞的迁移和侵袭能力。结论： 在MDA-MB-231细胞中,miR-21通过靶向调控PDCD4表达影响细胞的迁移和侵袭。miR-21可能成为抑制三阴性乳腺癌迁移和侵袭的靶点。     

miR-106b-5p靶向调控细胞周期蛋白D1抑制结直肠癌细胞增殖、迁移与侵袭

目的:探讨miR-106b-5p对结直肠癌(colorectal cancer,CRC)细胞增殖、迁移与侵袭的影响以及其作用机制.方法:实时荧光定量PCR检测miR-106b-5p在CRC组织与相应的癌旁组织、永生化的肠上皮细胞以及肠癌细胞中的表达量;CCK8实验检测DLD1细胞增殖能力;细胞划痕实验检测DLD1细胞的...     

miR-147a通过抑制CCND1和CDK4的表达对膀胱癌细胞周期和增殖的影响

目的:探讨miR-147a在膀胱癌组织、细胞中的表达及对膀胱癌细胞增殖和细胞周期的影响,研究miR-147a靶向负性调控CCND1和CDK4的表达对膀胱癌细胞周期和增殖的影响。方法:通过qRT-PCR检测miR-147a在正常膀胱组织和膀胱癌组织及正常膀胱上皮细胞、膀胱癌细胞系中的表达;使用miR-147a mimics转染观察细胞特性变化;CCK-8检测方法观察细胞增殖能力变化;流式细胞术观察细胞周期变化;生物信息学方法分析miR-147a的靶基因,并将靶基因进行GO分类和KEGG分析,找到与增殖和周期相关的基因;通过western blot方法验证miR-147a的靶基因,使用双萤光素酶报告基因系统检测miR-147a对CCND1和CDK4的转录活性影响;使用CCND1和CDK4特异性siRNA观察它们对细胞周期的影响,使用miR-147a inhibitor进行回补实验。结果:同正常组织/细胞相比,miR-147a在膀胱癌组织/膀胱癌细胞系中表达水平显著降低（P＜0.05）,miR-147a能够使膀胱癌细胞发生G1/S期阻滞,细胞增殖能力降低;通过一系列生信分析找到两个与细胞增殖和周期相关的miR-147a的靶基因CCND1和CDK4;通过间接和直接实验验证CCND1和CDK4是miR-147a的靶基因;下调CCND1和CDK4能够使膀胱癌细胞发生G1/S期阻滞,同时抑制miR-147a的表达能够解除G1/S期阻滞。结论:miR-147a作为一个抑癌因子,可作为诊断膀胱癌的生物标志物;miR-147a通过抑制CCND1和CDK4的表达抑制膀胱癌细胞周期G1/S期进程并抑制膀胱癌细胞增殖。     

MicroRNA-21 down-regulates the expression of tumor suppressor PDCD4 in human glioblastoma cell T98G

MicroRNAs have been linked to different cancer-related processes. The microRNA miR-21 appears to function as an anti-apoptosis factor in glioblastomas. However, the functional target genes of miR-21 are largely unknown in glioblastomas. In this study, bioinformatics analysis was used to identify miR-21 target sites in various genes. Luciferase activity assay showed that a number of genes involved in apoptosis, PDCD4, MTAP, and SOX5, carry putative miR-21 binding sites. Expression of PDCD4 protein correlates inversely with expression of miR-21 in a number of human glioblastoma cell lines such as T98G, A172, U87, and U251. Inhibition of miR-21 increases endogenous levels of PDCD4 in cell line T98G and over-expression miR-21 inhibits PDCD4-dependent apoptosis. Together, these results indicate that miR-21 expression plays a key role in regulating cellular processes in glioblastomas and may serve as a target for effective therapies.     

LINC00909靶向miR-365a-5p/FGFBP1分子轴调控结肠癌细胞增殖、迁移及侵袭的实验研究

目的 探讨LINC00909对结肠癌细胞增殖、迁移、侵袭的影响及可能机制.方法 购买正常结肠上皮细胞NCM460,结肠癌细胞HCT8、Caco-2及DLD-1;用实时荧光定量PCR(RT-qPCR)及蛋白质印迹法(Western blotting)检测NCM460细胞及结肠癌细胞中LINC00909、miR-365a-...     

miR-125a-3p靶向负调控PLK4抑制神经母细胞瘤细胞增殖及其机制探讨

目的:研究miR-125a-3p对神经母细胞瘤细胞增殖的影响,并探讨其作用机制。方法:运用qRT-PCR法检测Hela、SH-SY-5Y、SHEP、SK-N-BE细胞中miR-125a-3p的表达;将miR-125a-3p组（转染miR-125a-3p mimics）、miR-NC组（未转染细胞）、inhibitor-NC组（转染空inhibitor）、miR-125a-3p inhibitor组（转染miR-125a-3p inhibitor）、siPLK4组（转染siPLK4）、miR-125a-3p+Vector组（miR-125a-3p mimics和pcDNA 3.1共转染）、miR-125a-3p+PLK4组（miR-125a-3p mimics和pcDNA 3.1-PLK4共转染）以脂质体法转染至SH-SY-5Y细胞;MTT法检测各组细胞的增殖情况;Western blot检测各组细胞中PLK4、PIK3CA、Akt、p-Akt蛋白的表达;双荧光素酶报告基因检测实验检测各组细胞的荧光活性。结果:与Hela细胞相比,SH-SY-5Y、SHEP、SK-N-BE细胞中miR-125a-3p的表达均显著降低（P<0.05）;与Control组相比,miR-125a-3p组细胞的增殖显著降低,PLK4、PIK3CA、p-Akt蛋白的表达量均显著降低（P<0.05）;PLK4为miR-125a-3p的靶基因。过表达PLK4可逆转miR-125a-3p对SH-SY-5Y细胞增殖及PI3K/Akt信号通路的抑制作用。结论:miR-125a-3p可抑制神经母细胞瘤细胞增殖,其作用机制与靶向负调控PLK4有关,将可为神经母细胞瘤的治疗提供新靶点。     

miR103通过靶向PDCD10逆转顺铂诱导的前列腺癌细胞耐药

目的：探讨miR103在前列腺癌细胞耐药中的作用及机制。方法：RT－PCR检测miR103在人前列腺癌细胞系PC－3、LNCap、22Rv1和 DU145以及顺铂耐药细胞株 PC －3／DDP、LNCap／DDP、22Rv1／DDP和DU145／DDP中的表达。以PC－3／DDP细胞系为研究对象,采用miR103 mimic或inhibitor转染细胞,流式细胞仪检测细胞凋亡,Western blotting和RT－PCR检测PDCD10的表达。采用PDCD10过表达载体转染过表达miR103的细胞,流式细胞仪检测细胞凋亡,Western blotting和RT－PCR检测细胞P糖蛋白（P－gp）的表达。结果：与亲本细胞系比较,miR103在顺铂耐药细胞系中低表达。上调miR103,顺铂诱导的细胞凋亡显著增加;下调miR103,顺铂诱导的细胞凋亡减少,但不具有显著性。过表达miR103,PDCD10的表达显著减少。转染PDCD10过表达载体显著减少miR103 mimic诱导的PC－3／DDP细胞凋亡,逆转了miR103 mimic对P－gp表达的抑制作用。结论：miR103通过靶向PDCD10抑制P－gp的表达逆转前列腺癌细胞耐药。     

红景天苷通过miR-99a-5p/IGF-1R信号通路调节鼻咽癌细胞的增殖、凋亡、迁移和侵袭

目的:探讨红景天苷调节微小RNA-99a-5p（microRNA-99a-5p,miR-99a-5p）/胰岛素样生长因子1受体（insulin-like growth factor 1 receptor,IGF-1R）信号通路对鼻咽癌细胞增殖、凋亡、迁移和侵袭的影响。方法:实时荧光定量PCR（qRT-PCR）法检测人鼻咽上皮细胞（NP69）、鼻咽癌细胞系（CNE-2Z、HONE1、6-10B、HNE2）中miR-99a-5p、IGF-1R信使RNA（mRNA）表达水平;将对数期HNE2细胞分为对照组（常规培养HNE2细胞）、红景天苷低浓度组（含HNE2细胞的培养基中加入0.5 μg/mL的红景天苷）、红景天苷中浓度组（含HNE2细胞的培养基中加入1 μg/mL的红景天苷）、红景天苷高浓度组（含HNE2细胞的培养基中加入2 μg/mL的红景天苷）、模拟物（mimics）-NC组（HNE2细胞转染miR-99a-5p mimics阴性对照）、miR-99a-5p mimics组（HNE2细胞转染miR-99a-5p mimics）、红景天苷高浓度+抑制剂（inhibitor）NC组（含HNE2细胞的培养基中加入2 μg/mL的红景天苷并转染miR-99a-5p inhibitor阴性对照）、红景天苷高浓度+miR-99a-5p inhibitor组（含HNE2细胞的培养基中加入2 μg/mL的红景天苷并转染miR-99a-5p inhibitor）。qRT-PCR法检测各组HNE2细胞中miR-99a-5p、IGF-1R mRNA表达水平;细胞计数试剂盒-8、流式细胞术、Transwell小室、划痕实验、蛋白免疫印迹法分别检测HNE2细胞增殖能力、凋亡率、侵袭能力、迁移能力、IGF-1R蛋白表达水平;双荧光素酶报告基因实验验证miR-99a-5p与IGF-1R的靶向关系。结果:与NP69细胞相比,CNE-2Z、HONE1、6-10B、HNE2细胞系中miR-99a-5p表达水平降低,IGF-1R mRNA表达水平升高（P＜0.05）,其中HNE2细胞中miR-99a-5p、IGF-1R mRNA表达水平与NP69细胞差异更明显（P＜0.05）;与对照组相比,红景天苷低、中、高浓度组miR-99a-5p表达水平、增殖抑制率、凋亡率依次升高（P＜0.05）,IGF-1R mRNA与蛋白表达水平、侵袭细胞数目、划痕愈合率依次降低（P＜0.05）;与对照组、mimics-NC组相比,miR-99a-5p mimics组miR-99a-5p表达水平、增殖抑制率、凋亡率升高（P＜0.05）,IGF-1R mRNA与蛋白表达水平、侵袭细胞数目、划痕愈合率降低（P＜0.05）;与红景天苷高浓度组、红景天苷高浓度+inhibitor NC组相比,红景天苷高浓度+miR-99a-5p inhibitor组miR-99a-5p表达水平、增殖抑制率、凋亡率降低（P＜0.05）,IGF-1R mRNA与蛋白表达水平、侵袭细胞数目、划痕愈合率升高（P＜0.05）;miR-99a-5p与IGF-1R存在靶向关系。结论:红景天苷可能通过促进miR-99a-5p表达,靶向抑制IGF-1R表达,参与抑制HNE2细胞增殖、侵袭、迁移,并促进HNE2细胞凋亡。     

The microRNA-520a-3p inhibits proliferation,apoptosis and metastasis by targeting MAP3K2 in non-small cell lung cancer

Growing evidence indicates that miR-520a was involved in the complement attack and migration of tumor cells, but nonetheless, the role of miR-520a-3p in non-small cell lung cancer (NSCLC) is not clear. Mitogen-activated protein kinase kinase kinase 2 (MAP3K2) is a kinase belonging to the serine/threonine protein kinase family. To develop potential therapy targeting MAP3K2, we studied the roles of miR-520a-3p in the proliferation, apoptosis and metastasis of NSCLC. The expression levels of miR-520a-3p were quantified in tumor tissues of NSCLC by qRT-PCR, and the mimics and inhibitors were used to verify the function of miR-520a-3p. The cell proliferation was evaluated by MTT assay, and the migration and invasion was evaluated by transwell assay. The athymic mice subcutaneous injection was used to research NSCLC cell tumor formation. The bioinformatics tools and luciferase assay was applied to detect the relationship between miR-520a-3p and its target. Protein levels of miR-520a-3p target was determined by western blot analysis. MiR-520a-3p expression was decreased in the NSCLC tissues compared with their normal counterparts and lower expression of miR-520a-3p in NSCLC tissues was associated with a higher clinical stage, NSCLC metastasis and poor prognosis. Inhibition of expression of miR-520a-3p can reduce in vitro NSCLC cell migration and invasion as well as in vivo metastasis. MAP3K2 mRNA contains a binding site for miR-520a-3p in the 3’UTR. MAP3K2 is one of target of miR-520a-3p. Together, our data demonstrated that miR-520a-3p inhibits proliferation, apoptosis and metastasis in NSCLC by targeting MAP3K2, and miR-520a-3p may be used as a prognosis marker for NSCLC in clinical research.     

miR-203a-3p靶向AKT2增强骨肉瘤细胞的放射敏感性

目的:研究miR-203a-3p对骨肉瘤MG-63细胞凋亡和放射敏感性的影响及潜在作用机制.方法:qRT-PCR检测AKT2 mRNA的表达水平及不同放射剂量(0、2、4、6、8 Gy)照射后MG-63细胞中miR-203a-3p的表达水平,克隆形成实验检测不同放射剂量处理后细胞存活分数,流式细胞术测定MG-63细胞凋...     

miR-21调控程序性细胞死亡4基因的表达对宫颈癌细胞增殖和侵袭能力的影响

目的:探讨miR-21调控程序性细胞死亡4基因的表达对宫颈癌细胞增殖和侵袭能力的影响及其 机制。方法:qPCR检测miR-21和PDCD4在宫颈癌组织中的表达情况;双荧光素酶实验检测miR-21和PDCD4之间的调控作用;细胞集落实验检测抑制miR-21对宫颈癌细胞增殖能力的影响情况,Transwell侵袭实验检测对宫颈癌细胞侵袭能力的影响情况。结果:与癌旁正常组织相比,miR-21在宫颈癌组织中的表达上调,PDCD4在宫颈癌组织中表达下调;双荧光素酶实验检测miR-21和PDCD4之间的调控关系,miR-21可直接调控PDCD4蛋白的表达及活性情况;抑制miR-21的表达后可以抑制宫颈癌细胞的增殖能力;抑制miR-21的表达水平后,宫颈癌细胞的侵袭能力得到一定程度的抑制。结论:miR-21可以调控PDCD4的表达影响宫颈癌细胞的增殖和侵袭能力。     

胃癌患者血清miR-181d与PDCD4的表达及意义

 目的 分析胃癌患者血清miR-181d和PDCD4的表达及调控机制。方法 利用Real-time PCR检测6株胃黏膜细胞系中miR-181d和PDCD4的表达，并通过Western blot检测PDCD4的表达；采用Real-time PCR检测58例胃癌患者和50例健康体检者血清miR-181d的表达，并通过ELISA方法检测PDCD4的表达；采用化学法合成干扰/过表达miR-181d及构建PDCD4 3'-UTR荧光报告素载体检测miR-181d对PDCD4的调控。结果 RealtimePCR结果显示miR-181d和PDCD4在胃癌细胞系中存在差异表达；miR-181d在胃癌患者血清中的表达高于健康体检者（P<0.001），PDCD4表达则降低（P<0.05）, 通过临床相关性分析表明，胃癌血清miR-181d与PDCD4呈显著负相关（R²=-0.44），并在健康体检者中miR-181d与PDCD4蛋白呈负相关（R²=-0.426）；结合临床资料分析，血清miR-181d的表达与吸烟喝酒史呈正相关（P=0.015, P=0.034），而靶基因PDCD4仅与饮酒史相关（P<0.001）；ROC分析显示血清miR-181d与PDCD4用于体外诊断区分肿瘤和健康体检者具有良好的特异性与敏感度。过表达miR-181d于AGS细胞后PDCD4表达下降；反义miR-181d转染BGC823细胞后PDCD4表达增加。双荧光报告系统发现miR-181d可作用于PDCD4 3'-UTR区。结论 在胃癌患者血清中miR-181d的表达较高，而PDCD4的表达较低，并且miR-181d可能是通过作用于PDCD43'-UTR区发挥作用。     

MiR-138 induces cell cycle arrest by targeting cyclin D3 in hepatocellular carcinoma

The deregulation of microRNA (miRNA) is frequently associated with a variety of cancers, including hepatocellular carcinoma (HCC). In this study, we identified 10 upregulated miRNAs (miR-217, miR-518b, miR-517c, miR-520g, miR-519a, miR-522, miR-518e, miR-525-3p, miR-512-3p and miR-518a-3p) and 10 downregulated miRNAs (miR-138, miR-214, miR-214#, miR-27a#, miR-199a-5p, miR-433, miR-511, miR-592, miR-483-5p and miR-483-3p) by Taqman miRNAs array and quantitative real-time PCR (qRT-PCR) confirmation. Additionally, we investigated the expression and possible role of miR-138 in HCC. qRT-PCR results showed that miR-138 was downregulated in 77.8%(14/18) of HCC tissues compared with adjacent non-tumor tissues. Overexpression of miR-138 reduced cell viability and colony formation by induction of cell arrest in HCC cell lines and inhibited tumor cell growth in xenograft nude mice. The use of miR-138 inhibitor increased cell viability and colony formation in HCC cell lines and tumor cell growth in xenograft nude mice. Using TargetScan predictions, CCND3 was defined as a potential direct target of miR-138. Furthermore, CCND3 protein expression was observed to be negatively correlated with miR-138 expression in HCC tissues. The dual-luciferase reporter gene assay results showed that CCND3 was a direct target of miR-138. The use of miR-138 mimic or inhibitor could decrease or increase CCND3 protein levels in HCC cell lines. We conclude that the frequently downregulated miR-138 can regulate CCND3 and function as a tumor suppressor in HCC. Therefore, miR-138 may serve as a useful therapeutic agent for miRNA-based HCC therapy.     

肝癌细胞 miR-183表达及上下游调控机制探讨

目的： miR-183的显著上调已成为临床肝肿瘤发生的普遍特征,但其在肝癌发生中的分子机制亟待阐明。本研究探讨肝癌细胞 Hep3B 中 miR-183的表达及其上游调控机制和下游靶标蛋白变化。方法 MTT 法检测人正常肝细胞株 LO2及人肝癌细胞株 HepG2、Hep3B 的细胞增殖活性。提取细胞的总 RNA 和蛋白,采用 RT-qPCR 和蛋白印迹法分别检测细胞株中 miR-183的表达和细胞外信号调节激酶1／2（extracellular signal-regulated kinase 1／2,ERK1／2）、磷酸化 ERK1／2（phospho-ERK1／2,p-ERK1／2）、磷酸化胞内磷脂酰肌醇激酶（phospho-phosphatidylinositol-3-kinase,p-PI3K）、磷酸化蛋白激酶 B（phospho-protein kinase B,p-AKT）、NF-κB 抑制蛋白α亚基（nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor,alpha,IκBα）和程序性细胞死亡因子4（programmed cell death factor 4,PDCD4）的蛋白水平。用抑制剂分别抑制 Hep3B 细胞 ERK、PI3K、AKT 和 NF-κB 信号分子的活性并检测 miR-183的表达水平及 PD-CD4蛋白表达水平。结果 LO2、HepG2和 Hep3B 细胞在48 h 的增殖倍数分别为8．76±0．22、16．61±1．59和19．86±0．69,F ＝159．90,P ＜0．001。与 LO2（41．68±9．62）和 HepG2（41．53±1．20）细胞相比,Hep3B（69．15±11．02）的 miR-183细胞表达水平显著升高,F ＝10．250,P ＝0．012。与 LO2相比,Hep3B 细胞的 p-ERK1、ERK1、p-AKT 和 IκBα的蛋白水平明显升高,分别为 LO2的10．87、24．68、6．67和1．92倍;PDCD4的蛋白水平明显下降,为 LO2的0．14倍。与LO2细胞相比,HepG2细胞的 IκBα和 PDCD4蛋白表达明显升高,分别为 LO2的4．46和7．90倍。抑制 ERK 的活性后24 h,miR-183表达水平显著上升,F ＝215．459,P ＜0．001;抑制 Hep3B 细胞的 PI3K 活性后12和24 h,miR-183表达水平显著降低,F ＝80．215,P ＜0．001;抑制 AKT 的活性后12和24 h,miR-183表达水平显著下降,F ＝101．947,P ＜0．001;抑制 NF-κB 的活性后12和24 h,miR-183表达水平没有显著变化,F ＝1．826,P ＝0．216。抑制 ERK 和 NF-κB 信号分子活性对 PDCD4的蛋白表达没有显著影响。抑制 PI3K 和 AKT 信号分子活性可明显提升 PDCD4的蛋白表达水平。结论在 Hep3B 细胞中,PI3K／AKT 对 miR-183的表达有显著的促进作用,而 ERK 对 miR-183的表达有显著的抑制作用,NF-κB 不是调控 miR-183表达的主要信号分子。PDCD4是 miR-183的重要下游靶蛋白。     

miR-1297 inhibits osteosarcoma cell proliferation and growth by targeting CCND2

Cyclin D2 (CCND2) is abnormally overexpressed in many tumor types and has been associated with tumor cell proliferation. Although the important role of miR-1297 is well established, the molecular mechanism between CCND2 and miR-1297 in osteosarcoma (OS) has not been determined. In the present study, we found CCND2 was highly expressed in OS cells, and its downregulation suppressed cell proliferation, resulting in G1 phase cell cycle arrest. In contrast, miR-1297 was lowly expressed in OS compared to normal tissue. Several data platforms predicted that CCND2 was a target of miR-1297, which was validated by a dual-luciferase reporter assay that revealed miR-1297 could bind with CCND2-3’UTR. miR-1297 overexpression greatly inhibited CCND2 protein expression and exerted the same phenotypic effect as CCND2 downregulation in OS cells. Furthermore, miR-1297 inhibition could also be rescued by CCND2. Nude mice injected cells stable overexpressing miR-1297 OS cells showed lower size and tumor weight. Moreover, lower fluorescence activity recorded by in vivo imaging system and bone erosion revealed by microCT in the miR-1297 group demonstrated miR-1297 inhibited OS tumor growth via CCND2. Our findings demonstrated that miR-1297 can inhibit proliferation and tumor growth in OS by directly targeting CCND2, which indicates that miR-1297 may represent a novel therapeutic target for OS.     

lncRNA GAS6-AS2靶向miR-125a-3p调控肺癌A549细胞紫杉醇敏感性

目的:探讨长链非编码RNA(long non-coding RNA,lncRNA)GAS6-AS2对肺癌A549细胞增殖、迁移、侵袭及紫杉醇(paclitaxel,PTX)敏感性的影响及其分子机制.方法:用qPCR法检测肺癌A549和A549/PTX细胞中GAS6-AS2和miR-125a-3p 的表达水平.用脂质体转...     

长链非编码RNA SNHG1靶向miR-340-5p/CCND1轴调控食管癌细胞增殖、迁移和侵袭

目的:分析长链非编码RNA小核仁RNA宿主基因1（LncRNA SNHG1）靶向miR-340-5p/细胞周期蛋白1（CCND1）轴调控食管癌细胞增殖、迁移和侵袭。方法:体外培养人食管上皮细胞、食管癌细胞KYSE-30、TE-1、NEC、Eca109,qRT-PCR法测定细胞中LncRNA SNHG1、miR-340-5p、CCND1 mRNA水平。将对数期NEC细胞分为对照组、sh NC1组、sh LncRNA SNHG1组、sh NC2组、sh CCND1组、sh LncRNA SNHG1+miR-340-5p inhibitor组、sh CCND1+miR-340-5p inhibitor组。CCK-8法测定细胞增殖能力,Transwell小室法测定细胞侵袭、迁移能力,Western blot法检测CCND1、Ki-67、MMP-2蛋白表达,双荧光素酶验证miR-340-5p与LncRNA SNHG1、CCND1的靶向关系,通过裸鼠瘤内注射转染试剂进行体内试验。结果:与人食管上皮细胞相比,食管癌细胞KYSE-30、TE-1、NEC、Eca109中LncRNA SNHG1、CCND1 mRNA表达升高,miR-340-5p表达降低（P＜0.05）,其中NEC细胞变化最显著,所以使用NEC作为下续研究菌株。与对照组、sh NC组相比,sh LncRNA SNHG1组NEC细胞LncRNA SNHG1、OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2降低（P＜0.05）;与对照组、sh NC组相比,sh CCND1组CCND1 mRNA与蛋白表达、OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2表达降低（P＜0.05）。miR-340-5p与LncRNA SNHG1、CCND1均靶向结合,与sh LncRNA SNHG1组相比,sh LncRNA SNHG1+miR-340-5p inhibitor组OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2蛋白表达升高（P＜0.05）;与sh CCND1组相比,sh CCND1+miR-340-5p inhibitor组OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2蛋白表达升高（P＜0.05）;裸鼠移植瘤实验进行了体内验证。结论:LncRNA SNHG1沉默可能通过调控miR-340-5p/CCND1表达抑制食管癌NEC细胞增殖、侵袭与迁移,裸鼠体内也验证了这一结果。