miR-212 promotes pancreatic cancer cell growth and invasion by targeting the hedgehog signaling pathway receptor patched-1

Background

microRNAs (miRNAs) are a class of small non-coding RNAs that play important roles in carcinogenesis. In the present study, we investigated the effect of miR-212 on pancreatic ductal adenocarcinoma (PDAC) and its target protein.

Methods

Quantitative real-time PCR(qRT-PCR) was performed to detect the expression of miR-212 in PDAC tissues and pancreatic cancer cell lines. miR-212 mimic, miR-212 inhibitor and negative control were transfected into pancreatic cancer cells and the effect of miR-212 up-regulation and down-regulation on the proliferation, migration and invasion of cells were investigated. Furthermore, the mRNA and protein levels of Patched-1(PTCH1) were measured. Meanwhile, luciferase assays were performed to validate PTCH1 as miR-212 target in PDAC.

Results

miR-212 was up-regulated in PDAC tissues and cells.Using both gain-of function and loss-of function experiments, a pro-oncogenic function of miR-212 was demonstrated in PDAC. Moreover, up-regulated of PTCH1 could attenuate the effect induced by miR-212.

Conclusion

These data suggest that miR-212 could facilitate PDAC progression and metastasis through targeting PTCH1, implicating a novel mechanism for the progression of PDAC.     

LINC02163靶向miR-338-3p影响乳腺癌细胞增殖、侵袭及迁移

背景与目的:长链非编码RNA(long non-coding RNA,lncRNA)已被发现在乳腺癌中失调,与肿瘤恶性行为密切相关。本研究旨在探究LINC02163靶向miR-338-3p对乳腺癌细胞增殖、侵袭及迁移的影响。方法:收集郑州人民医院2020年1月—2021年9月收治的9例乳腺癌患者的乳腺癌组织及距其2 cm外的癌旁组织样本,通过实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测组织样本、人正常乳腺上皮细胞系(MCF-10A)和乳腺癌细胞系(MCF-7、BT-20、MDA-MB-231、T47D)中LINC02163的表达。将MDA-MB-231分为control组、sh-NC组、sh-LINC02163组、sh-LINC02163+inhibitor-NC组和sh-LINC02163+miR-338-3p inhibitor组。采用MTT法、transwell实验及划痕实验分别检测MDA-MB-231细胞活力、侵袭及迁移能力。采用蛋白质印迹法(West...     

Upregulated fascin1 in non-small cell lung cancer promotes the migration and invasiveness,but not proliferation

Fascin1, an actin-binding protein, is overexpressed in many kinds of tumors. However, its exact role in non-small cell lung cancer (NSCLC) is still unclear. We observed that expression of fascin1 was associated with lymph nodes metastasis and TNM staging but not proliferation in NSCLC by using immunohistochemistry. By generating A549 lung cancer clones stably overexpressing different fascin1 protein, we further confirmed that fascin1 could promote A549 cell migration and invasiveness in vitro and in vivo. Using cell proliferation assay, cell cycle analysis in vitro and tumorigenicity assay in vivo, we showed that fascin1 had little influence on proliferation in A549 cells. These results suggest that fascin1 plays a significant role in NSCLC metastasis and may be a target of intervening to prevent the metastasis of NSCLC.     

MTA1 promotes the invasion and migration of non-small cell lung cancer cells by downregulating miR-125b

Background

The metastasis-associated gene 1 (MTA1) has been identified as one critical regulator of tumor metastasis. Previously, we identified miR-125b as a downregualted miRNA in non-small cell lung cancer (NSCLC) cell line upon MTA1 depletion. However, the role of miR-125b and MTA1 in the regulation of NSCLC metastasis remains unclear.

Methods

Stable MTA1 knockdown NSCLC cell lines 95D and SPC-A-1 were established by transfection with MTA1 shRNA. The effects of MTA1 depletion on the expression of miR-125b and cell migration and invasion were examined by real-time PCR, wound healing and matrigel invasion assay.

Results

MTA1 knockdown led to the upregulation of miR-125b level in NSCLC cells. Furthermore, MTA1 knockdown reduced while miR-125b inhibitor enhanced cell migration and invasion of NSCLC cells. Notably, miR-125b inhibitor antagonized MTA1 siRNA induced inhibition of cell migration and invasion.

Conclusion

MTA1 and miR-125b have antagonistic effects on the migration and invasion of NSCLC cells. The newly identified MTA1-miR-125b axis will help further elucidate the molecular mechanism of NSCLC progression and suggest that ectopic expression of miR-125b is a potentially new therapeutic regimen against NSCLC metastasis.     

Apolipoprotein E expression promotes lung adenocarcinoma proliferation and migration and as a potential survival marker in lung cancer

Many human lung cancer cell lines express apolipoprotein E (ApoE), especially cells derived from malignant pleural effusions (MPE) in patients with lung adenocarcinoma. This study aimed to investigate the influence of ApoE expression on lung cancer. In lung cancer tissues, ApoE expression was more frequently found in malignant pleural effusions (MPE)-associated lung adenocarcinoma than in lung adenocarcinoma or squamous cell carcinoma without MPE (P < 0.05), indicating that ApoE is associated with the pathogenesis of MPE in patients with lung adenocarcinoma. Next, we examined the roles of ApoE in an MPE-derived lung adenocarcinoma cell line that endogenously over-expresses ApoE, PC14PE6/AS2 (AS2). In that experiment we inhibited ApoE expression by transfection of a plasmid carrying ApoE siRNAs into AS2 cells to generate AS-S2 and AS-S3 cells. Compared to vector-control cells and parental AS2 cells, AS2-S2 and AS2-S3 cells grew slower (P < 0.05), were more sensitive to cisplatin, and had significantly impaired cellular migration (P < 0.05). Furthermore, over-expression of ApoE was independently associated with poor survival in lung adenocarcinoma patients who had MPE at the time of diagnosis (P < 0.001). Conclusively, ApoE over-expression promotes cancer proliferation and migration and contributes to an aggressive clinical course in patients with lung adenocarcinoma and MPE.     

miR-625-5p通过靶向调控PRKACA促进肺腺癌细胞的增殖和侵袭

背景 与目的:肺腺癌是非小细胞肺癌的一种亚型,虽在诊断和治疗方面已经取得了很大进展,但晚期肺腺癌临床预后和总生存仍较差.近年来多项研究表明,miRNA在多种癌症中发挥作用,并在细胞增殖、转移、炎症等生物学过程中发挥重要作用.探究miR-625-5p对肺腺癌细胞增殖和侵袭能力的影响及分子机制,旨在为后续肺腺癌的诊断和治疗...     

MicroRNA-106b promotes colorectal cancer cell migration and invasion by directly targeting DLC1

Background

Growing evidence suggests that microRNAs (miRNAs) play an important role in tumor development, progression and metastasis. Aberrant miR-106b expression has been reported in several cancers. However, the role and underlying mechanism of miR-106 in colorectal cancer (CRC) have not been addressed.

Methods

Quantitative RT-PCR(qRT-PCR) was performed to evaluate miR-106b levels in CRC cell lines and patient specimens. Cell proliferation was detected using MTT assay, and cell migration and invasion ability were evaluated by wound healing assay and transwell assay. The target gene of miR-106b was determined by qRT-PCR, western blot and luciferase assays.

Results

miR-106b was significantly up-regulated in metastatic CRC tissues and cell lines, and high miR-106b expression was associated with lymph node metastasis and advanced clinical stage. In addition, miR-106b overexpression enhances, whereas miR-106b depletion reduces CRC cell migration and invasion. Moreover, we identify DLC1 as a direct target of miR-106b, reveal its expression to be inversely correlated with miR-106b in CRC samples and show that its re-introduction reverses miR-106b-induced CRC cell migration and invasion. Furthermore, survival analyses showed the patients with high mi-106b/low DLC1 had shorter overall survival (OS) and disease-free survival (DFS) rates, and confirmed miR-106b may be an independent prognostic factor for OS and DFS in CRC patients.

Conclusions

Our findings indicate that miR-106b promotes CRC cell migration and invasion by targeting DLC1. This miRNA may serve as a potential prognostic biomarker and therapeutic target for CRC.     

miR-145靶向KLF5抑制膀胱癌BIU-87细胞增殖、侵袭、迁移的机制研究

目的 研究miR-145靶向调控锌指蛋白转录因子5(KLF5)抑制膀胱癌BIU-87细胞增殖、侵袭、迁移的机制.方法 miR-145 mimics对BIU-87细胞进行转染,实验分为对照组、miR-145阴性对照组、miR-145 mimics组,SV-HUC-1细胞作为SV-HUC-1组.实时荧光定量法(RT-qPC...     

miR⁃93⁃5p靶向PKMYT1对肺腺癌A549细胞增殖、迁移及侵袭的影响

目的 探讨miR⁃93⁃5p靶向蛋白激酶膜相关酪氨酸/苏氨酸1（protein kinase membrane associated tyrosine/threonine 1,PKMYT1）对肺腺癌细胞增殖、迁移及侵袭的影响及其作用机制。方法 使用LipofectamineTM 2000分别将质粒inhibitor NC、miR⁃93⁃5p inhibitor、pcDNA、pc⁃PKMYT1转染至A549细胞中,记为inhibitor NC组、miR⁃93⁃5p inhibitor组、pcDNA组和pc⁃PKMYT1组,同时将未转染的细胞记为control组。利用qRT⁃PCR检测细胞中miR⁃93⁃5p、PKMYT1 mRNA的表达水平。采用双荧光素酶报告基因实验验证miR⁃93⁃5p与PKMYT1的靶向关系;CCK⁃8法检测细胞增殖能力;流式细胞术检测细胞周期情况;Transwell实验检测细胞迁移和侵袭能力;Western blot法检测细胞中PKMYT1、细胞周期素D1（Cyclin D1）、细胞周期素B1（Cyclin B1）的蛋白表达水平。结果 与BEAS⁃2B细胞相比,SPC⁃A⁃1细胞、A549细胞、LTEP⁃α⁃2细胞中miR⁃93⁃5p表达水平均升高（均P<0.01）,PKMYT1 mRNA和蛋白表达水平均降低（均P<0.001）。转染24 h后,分别与control组、inhibitor NC组相比,miR⁃93⁃5p inhibitor组A549细胞增殖活性、S期细胞比例、迁移细胞数目、侵袭细胞数目、Cyclin D1和Cyclin B1蛋白水平均降低（均P<0.05）,G2/M期细胞比例升高（P<0.001）。双荧光素酶报告基因实验结果显示,与miR⁃93⁃5p NC+3'UTR⁃WT组相比,miR⁃93⁃5p mimic+3'UTR⁃WT组细胞相对荧光素酶活性下降（P<0.001）。转染24 h后,与pcDNA组相比,pc⁃PKMYT1组A549细胞增殖活性、S期细胞比例、迁移细胞数目、侵袭细胞数目、Cyclin D1和Cyclin B1蛋白水平均降低（均P<0.001）,G2/M期细胞比例升高（P<0.001）。结论 沉默miR⁃93⁃5p可能通过上调PKMYT1表达水平抑制肺腺癌细胞增殖、迁移及侵袭。     

miR-758-3p通过靶向核仁纺锤体相关蛋白1抑制非小细胞肺癌细胞增殖和侵袭

目的探讨miR-758-3p在非小细胞肺癌(NSCLC)中的生物学功能及其作用机制。方法采用脂质体法转染miR-758-3p模拟物(miR-758-3p模拟物组)、miRNA对照(对照组)、核仁纺锤体相关蛋白1(NUSAP1)过表达质粒(NUSAP1过表达组)、miR-758-3p模拟物+NUSAP1(miR-758-3p模拟物+NUSAP1组)至人NSCLC细胞系A549细胞中。采用CCK-8法和Transwell小室法检测A549细胞增殖、迁移和侵袭能力,结合生物信息学预测网站和双荧光素酶报告基因实验验证miR-758-3p及其靶基因的靶向关系。结果转染24 h后,miR-758-3p模拟物组和对照组A549细胞中miR-758-3p基因的相对表达水平分别为3.02±0.16和1.00±0.08,NUSAP1基因的相对表达水平分别为0.04±0.02和1.00±0.03,差异均有统计学意义(均P<0.05)。转染72 h后,miR-758-3p模拟物组和对照组细胞的吸光度值分别为0.78±0.06和1.15±0.06,差异有统计学意义(P<0.05)。miR-758-3p模拟物组细胞的迁移数和侵袭数分别为[(119.04±11.49)个]和[(71.33±5.36)个],均低于对照组[分别为(271.38±19.05)个和(164.30±8.11)个;均P<0.05]。miR-758-3p模拟物转染野生型NUSAP1报告基因的荧光素酶活性(0.37±0.04)低于对照组(1.00±0.03,P<0.05)。突变型NUSAP1报告基因和对照组的荧光素酶活性分别为0.96±0.02和0.95±0.02,差异无统计学意义(P>0.05)。结论miR-758-3p通过调控NUSAP1基因的表达抑制NSCLC细胞的增殖、迁移和侵袭。     

LINC00243靶向miR-1976促进甲状腺癌细胞增殖、迁移和侵袭

目的:探讨长链非编码RNA（lncRNA）LINC00243对甲状腺癌细胞增殖、迁移及侵袭的影响和分子机制。方法:实时荧光定量PCR（qRT-PCR）检测正常甲状腺细胞系（HT-ori3）和甲状腺癌细胞系（BCPAP、TPC-1和SW1736）中LINC00243和miR-1976的表达水平。将LINC00243小干扰RNA（si-LINC00243）、miR-1976模拟物（miR-1976 mimics）分别转染TPC-1细胞。细胞计数试剂盒（CCK-8）检测细胞活力;Transwell实验检测细胞迁移和侵袭数量;蛋白质印记（Western blot）检测细胞周期素D1（CyclinD1）、基质金属蛋白酶2（MMP2）和MMP9的表达水平。双荧光素酶报告基因实验和qRT-PCR验证LINC00243和miR-1976的靶向调控关系。结果:与HT-ori3细胞比较,3种甲状腺癌细胞中LINC00243的表达水平显著升高,miR-1976的表达水平显著降低。沉默LINC00243或高表达miR-1976均可抑制TPC-1细胞的增殖、迁移和侵袭,抑制CyclinD1、MMP2和MMP9蛋白的表达（P＜0.05）。LINC00243靶向负性调控miR-1976表达。低表达miR-1976可逆转沉默LINC00243对TPC-1细胞增殖、迁移和侵袭的抑制作用（P＜0.05）。结论:在甲状腺癌细胞中,LINC00243呈高表达,miR-1976呈低表达。LINC00243通过靶向调控miR-1976促进甲状腺癌细胞增殖、迁移和侵袭。     

miR-491-5p靶向FGFR4调节肝母细胞瘤细胞增殖和迁移

目的 探讨miR-491-5p靶向调节FGFR4对肝母细胞瘤增殖和迁移的影响及可能的作用机制。方法 采用qRT-PCR检测15例小儿肝母细胞瘤组织及其癌旁正常组织,正常肝细胞LO2和肝母细胞瘤细胞HuH-6中miR-491-5p和FGFR4 mRNA的表达;Western blot检测FGFR4蛋白表达;TargetScan数据库预测miR-491-5p靶点;双荧光素酶报告基因实验检测miR-491-5p与FGFR4的靶向关系。将mimic NC、miR-491-5p mimic、pcDNA3.1和pcDNA3.1-FGFR4质粒分别转染至HuH-6细胞后,采用qRT-PCR与Western blot检测FGFR4的表达,CCK-8实验检测细胞增殖能力,Transwell小室实验检测细胞迁移能力,Western blot检测GSK3β/β-catenin信号通路相关蛋白的表达。结果 小儿肝母细胞瘤组织和HuH-6细胞中miR-491-5p表达水平均低于癌旁正常组织及肝细胞LO2（P<0.05）,而FGFR4表达水平高于癌旁正常组织及肝细胞LO2（P<0.05）;TargetScan数据库预测显示miR-491-5p与FGFR4存在结合位点。与mimic NC组比较,过表达miR-491-5p可抑制FGFR4表达、HuH-6细胞增殖和迁移能力及p-GSK3β/GSK3β、p-β-catenin/β-catenin及p-C-myc/C-myc比值（均P<0.05）。与mimic+Pc组比较,过表达FGFR4可逆转过表达miR-491-5p对HuH-6细胞增殖、迁移及p-GSK3β/GSK3β、p-β-catenin/β-catenin和p-C-myc/C-myc比值的抑制作用（均P<0.05）。结论 miR-491-5p通过负向调控FGFR4抑制肝母细胞瘤细胞增殖和迁移,可能与GSK3β/β-catenin信号通路失活有关。       

miR-572 通过靶向调控 WWOX 影响肺癌细胞恶性生物学行为

目的 探讨miR-572靶向氧化还原酶的WW结构域（WW domain containing oxidoreductase,WWOX）调控肺癌细胞增殖、凋亡的分子机制。方法 选取50例2016年3月—2018年5月十堰市太和医院手术切除肺癌组织及其相应的癌旁组织。利用脂质体转染技术将miR-572 inhibitor和miR-572 mimics转染至肺癌A549和L9981细胞;采用qRT-PCR检测miR-572和WWOX的mRNA表达水平,MTT法检测细胞增殖能力,流式细胞仪检测细胞周期和细胞凋亡情况。通过生物信息学软件Targetscan分析miR-572的靶基因,荧光素酶报告基因实验检验miR-572和WWOX的靶向关系。结果 miR-572在肺癌组织和细胞中高表达（均P<0.05）。下调miR-572后,肺癌细胞A549、L9981中miR-572表达水平均降低（均P<0.05）,细胞增殖能力也降低（均P<0.05）,细胞G0/G1期比例和细胞凋亡率均升高（均P<0.05）;而上调miR-572后,肺癌A549、L9981细胞增殖能力升高（均P<0.05）,细胞G0/G1期比例和细胞凋亡率则降低（均P<0.05）。WWOX在肺癌组织和细胞中低表达（均P<0.05）,且在肺癌组织中WWOX与miR-572表达呈负相关（r=-0.669,P<0.001）。下调WWOX可逆转下调miR-572对肺癌细胞的增殖抑制、周期阻滞和凋亡促进作用。 结论 下调miR-572可抑制肺癌细胞增殖并诱导细胞凋亡,其作用机制可能与靶向负调控WWOX有关。     

lncRNA BLACAT1靶向LEMD1促进结直肠癌细胞的增殖和转移

目的 研究长链非编码RNA(lncRNA)BLACAT1在结直肠癌细胞发生发展过程的作用机制。方法 starBase分析TCGA数据库中lncRNA BLACAT1分别在结直肠癌组织和癌旁组织中的表达水平,采用qPCR分别检测10例结直肠癌患者癌组织和癌旁组织中lncRNA BLACAT1的表达水平;检测结直肠癌细胞系SW620、SW480、HT-29、LoVo、HCT116和正常结直肠上皮细胞FHC中lncRNA BLACAT1的mRNA表达水平,筛选lncRNA BLACAT1高表达细胞系;敲低lncRNA BLACAT1验证对高表达细胞系增殖、迁移和侵袭的影响;starBase在线预测与lncRNA BLACAT1靶向作用的基因,qPCR和Western blot实验验证结果;starBase分析lncRNA BLACAT1与作用基因相关性,双荧光素酶报告基因实验验证lncRNA BLACAT1的靶基因;检测lncRNA BLACAT1作用靶基因对结直肠癌细胞的增殖、迁移和侵袭的影响。结果 TCGA数据库分析和结直肠癌患者检测结果均表明癌组织中lncRNA BLACAT1表达明显高于癌旁组织(P＜0.001);结直肠癌细胞系SW620中lncRNA BLACAT1的mRNA表达水平最高(P＜0.001);敲低lncRNA BLACAT1能够抑制SW620细胞的增殖(P＜0.001)、迁移(P＜0.001)和侵袭能力(P＜0.001);qPCR和Western blot实验及数据库分析结果表明lncRNA BLACAT1与LEMD1的表达存在正相关关系(r=0.778,P＜0.001),双荧光素酶报告基因实验证明LEMD1是lncRNA BLACAT1的靶基因;lncRNA BLACAT1上调LEMD1的表达,促进SW620细胞的增殖(P＜0.001)、迁移(P＜0.001)和侵袭(P＜0.001)。结论 lncRNA BLACAT1靶向LEMD1促进结直肠癌细胞SW620的增殖和转移。     

NOP14 promotes proliferation and metastasis of pancreatic cancer cells

Pancreatic cancer (PC) is one of the most malignant cancers worldwide. We describe a novel gene, NOP14, which plays significant roles in PC cell proliferation and migration. Inhibition or overexpression of NOP14 expression in PC cells can reduce or promote motility, proliferation and metastatic capacity in vivo. In parallel, we observed changes in proteins related to migration, such as E-cadherin, vimentin, MMP9, RhoA and p53, along with proteins involved in proliferation, such as MAPK3 and CDK2. Taken together, our study provides new evidence for NOP14 in regulating PC cell proliferation and migration, and may provide new insights for clinical diagnosis and therapy.     

MicroRNA-150对上皮性卵巢癌细胞增殖、凋亡和侵袭转移能力的影响

目的 研究microRNA-150(miR-150)在人上皮性卵巢癌细胞中的表达情况,及其对人上皮性卵巢癌细胞增殖、凋亡和侵袭转移能力的影响。方法 通过荧光实时定量PCR(qRT-PCR)检测各处理组细胞中的miR-150表达水平;利用MTT法、流式细胞技术、Transwell法探究miR-150的表达对上皮性卵巢癌细胞增殖、凋亡及侵袭转移能力的影响。结果 与正常卵巢上皮细胞(T29)相比,上皮性卵巢癌细胞(A2780和OVCAR3)中miR-150表达显著降低(P＜0.01);转染miR-150mimic后,A2780和OVCAR3细胞中miR-150水平明显升高(P＜0.01);MTT试验显示,转染培养三天后,miR-150 mimic组细胞OD值(A2780:1.12±0.03;OVCAR3:1.91±0.03)较Blank组(A2780:2.35±0.09;OVCAR3:2.63±0.07)及miR-150 NC组(A2780:2.18±0.07;OVCAR3:2.43±0.11)明显降低(P＜0.01);细胞凋亡检测显示:miR-150 mimic组凋亡率(A2780:16.10±0.58%;OVCAR3:15.16±1.30%)较Blank组(A2780:10.07±0.66%;OVCAR3:3.81±0.24%)及miR-150 NC组(A2780:10.36±1.08%;OVCAR3:4.89±0.07%)明显升高(P＜0.01);Transwell试验显示:miR-150 mimic组穿膜细胞数(A2780:38.67±2.03;OVCAR3:28.67±2.03)较Blank组(A2780:76.30±7.45;OVCAR3:55.67±3.18)及miR-150 NC组(A2780:74.33±5.78;OVCAR3:56.33±3.84)明显减少(P＜0.01)。结论 miR-150在上皮性卵巢癌细胞中表达降低,可能是上皮性卵巢癌增殖、侵袭转移的机制之一;上调miR-150的表达可以抑制上皮性卵巢癌细胞增殖、促进细胞凋亡,并且降低上皮性卵巢癌细胞侵袭转移能力。     

miR-203下调Bmi-1基因对肺腺癌细胞侵袭转移的影响

目的 探讨miR-203在肺腺癌中的表达,并分析其与肺腺癌细胞侵袭转移的关系及其分子机制。方法 实时定量PCR检测40例肺腺癌患者肿瘤组织中miR-203的相对表达水平及其与临床病理特征之间的关系;实时定量PCR检测H1650、A549、H1975、SPC-A-1肺腺癌细胞株中miR-203表达水平;生物信息学软件预测miR-203潜在的靶基因;脂质体2000介导miR-203模拟物、Bmi-1基因或Bmi-1 siRNA转染H1975细胞株;Western blotting检测Bmi-1蛋白水平;双荧光素酶报告基因验证miR-203是否作用于Bmi-1 mRNA的3’UTR 区预测靶位;Transwell小室侵袭实验检测H1975细胞株的侵袭转移能力。结果 40例肺腺癌组织中miR-203的相对表达量为0.065±0.013;肺腺癌miR-203表达与淋巴结转移有关,而与其他临床病理参数均无关;miR-203在肺腺癌细胞株H1650、A549、H1975、SPC-A-1中的相对表达量分别为0.280±0.102、0.308±0.168、0.167±0.073和0.287±0.096。生物信息学软件预测Bmi-1是miR-203的潜在靶基因;过表达miR-203可明显降低Bmi-1蛋白表达水平;双荧光素酶报告基因检测证明miR-203可作用于Bmi-1基因mRNA的 3’UTR区预测靶位。过表达miR-203+Bmi-1 siRNA可显著抑制肺腺癌细胞株H1975的侵袭迁移能力;在miR-203过表达的H1975细胞株中同时过表达Bmi-1可恢复其侵袭能力。结论 miR-203可通过下调Bmi-1基因表达抑制H1975肺腺癌细胞株的侵袭转移,是一种潜在抑制转移的miRNA分子。     

MicroRNA-195 inhibits non-small cell lung cancer cell proliferation,migration and invasion by targeting MYB

MicroRNA-195 (miR-195) has been implicated in several other cancers; however, its role in non-small cell lung cancer (NSCLC) remains unclear. In this study, we demonstrated that miR-195 was significantly down-regulated in NSCLC samples and cell lines compared with corresponding normal counterparts. In vitro and in vivo functional assays demonstrated that modulation of miR-195 expression affected NSCLC cell proliferation, migration and invasion. Using miRNA target prediction algorithms and reporter assays, we demonstrated that miR-195 suppressed the expression of MYB both at the mRNA and protein level, and was directly bound to the 3′untranslated region of MYB mRNA. Overexpression of MYB in NSCLC cells using an ectopic expression vector restored the decreased cell proliferation, migration and invasion effects induced by miR-195. Finally, we observed an inverse correlation between MYB and miR-195 in NSCLC. Taken together, our findings indicated that miR-195 functions as tumour suppressor in NSCLC, and the miR-195/MYB axis might represent a potential therapeutic target for NSCLC intervention.     

miR-200a抑制YAP1基因表达对肺癌细胞增殖的影响

  目的   研究微小RNA-200a（miR-200a）对肺癌细胞增殖的影响,并探讨其分子机制。   方法   采用Real-time PCR检测15例非小细胞肺癌组织和对应癌旁组织、人肺癌细胞株（A549、NCI-H520、SK-MES-1）及人正常肺支气管上皮细胞株16HBE中miR-200a的表达水平。用CCK-8法检测miR-200a对A549肺癌细胞增殖活性的影响。通过生物信息学方法预测miR-200a可能的靶基因,双荧光素酶报告基因实验结合Real-time PCR和Western blot验证miR-200a对靶基因YAP1的调控作用。CCK-8法检测下调靶基因YAP1对A549肺癌细胞株增殖活性的影响。   结果   miR-200a在非小细胞肺癌组织和肺癌细胞系中表达明显降低（P < 0.01）。上调miR-200a表达后明显抑制A549肺癌细胞的增殖活力（P < 0.01）。双荧光素酶报告基因显示miR-200a可以直接作用于靶基因YAP1的3'-UTR区域抑制荧光素酶活性（P < 0.01）,Real-time PCR和Western blot检测显示上调miR-200a的表达能够明显下调A549肺癌细胞YAP1 mRNA和蛋白的表达水平（P < 0.01）。CCK-8法显示下调YAP1的表达能够明显抑制A549肺癌细胞的增殖活性（P < 0.01）。   结论   miR-200a通过靶向作用于YAP1基因来抑制肺癌细胞的增殖,从而在肺癌中发挥抑癌基因的功能。