反义miR-21抑制宫颈鳞状细胞癌CaSki细胞系生长体外研究

目的探讨敲低miR-21表达对宫颈鳞状细胞癌细胞系CaSki增殖活性与凋亡的影响。方法实时荧光定量RT-PCR法检测正常宫颈组织、CaSki细胞转染前后miR-21的表达水平,采用MTT法、流式细胞术等技术检测转染后CaSki细胞增殖活性与凋亡的变化。结果 miR-21在CaSki细胞中表达水平为正常宫颈组织的3.25倍。CaSki细胞中转染反义miR-21(AS-miR-21)可显著抑制其表达。转染后24h、48h、72h、96h,MTT检测提示10nmol与20nmolAS-miR-21转染组中细胞增殖率较对照组明显降低(P<0.05),自48h抑制作用显现,且随着转染浓度增加,抑制作用增强,呈现剂量依赖性。细胞周期检测结果显示AS-miR-21转染组的G0/G1期细胞明显增多,S期细胞减少(P<0.05)。AnnexinⅤ联合PI技术检测细胞凋亡,结果显示AS-miR-21转染组早期凋亡细胞率[(23.40±2.16)%]明显高于空白对照组[(5.1±2.16)%]、阴性对照组[(6.87±0.55)%](P=0.000)。结论 AS-miR-21转染可敲低CaSki细胞中miR-21表达,并有效抑制细胞增殖活性,促进细胞凋亡。     

反义miR-21对荷人宫颈鳞癌裸鼠移植瘤生长和凋亡的影响

目的 构建荷人宫颈鳞癌裸鼠移植瘤模型,探讨干扰miR-21 表达对移植瘤生长、凋亡的影 响.方法 将人工合成的反义寡核苷酸ASODN(AS-miR-21)转染SiHa 细胞(抑制组),同时设阴性对照组和 空白对照组,对数生长期的SiHa 细胞分别接种于裸鼠皮下,绘制肿瘤生长曲线并计算肿瘤生长率.当肿瘤 体积达0.2 cm3 时采用AS-miR-21 多点瘤周注射,观察其对移植瘤的影响.应用免疫组化技术、荧光TUNEL 检测移植瘤细胞增殖活性和凋亡情况.结果 (1)成功构建人宫颈鳞癌裸鼠皮下移植瘤模型,抑制组、阴性 对照组及空白对照组成瘤率分别为37.5%、75.0%、100.0%,瘤体平均体积为(732.80 ±56.32 ) mm3 、(1228.46 ±78.53)mm3 、(1301.26 ±80.63)mm3,平均生长率为18.32%、30.71%和32.53%,瘤重为(0.73 ± 0.05)g、(1.26 ±0.12)g 和(1.35 ±0.25)g,抑制组与阴性、空白对照组比较差异均有统计学意义(P <0.05); (2)免疫组化显示抑制组裸鼠瘤体Ki67 表达显著下降;荧光TUNEL 凋亡检测显示抑制组裸鼠肿瘤细胞凋亡 率明显增多;(3)注射治疗剂量AS-miR-21 两周后观察肿瘤组织局部坏死严重,胞核裂解、消失,凋亡明显增 加.结论 AS-miR-21 可下调裸鼠人宫颈鳞癌移植瘤中miR-21 的表达,抑制移植瘤生长和促进其凋亡.     

miRNA-21siRNA对宫颈癌Hela细胞周期及细胞凋亡的影响

目的探讨microRNA-21(miR-21)siRNA对宫颈癌Hela细胞周期和细胞凋亡的影响。方法实时荧光定量PCR检测Hela细胞转染后siRNA干扰组及阴性对照组中miR-21的表达量。流式细胞仪分析Hela细胞周期及凋亡率的变化。结果转染siRNA干扰载体后,实时荧光定量PCR检测Hela细胞中miR-21明显低于空白对照组及阴性对照组,差异有统计学意义(P0.05),空白对照组与阴性对照组相比,差异无统计学意义(P0.05);siRNA表达载体可明显增加细胞凋亡率,并显现出G_0/G_1期阻滞作用。结论miR-21siRNA干扰载体可以高效地抑制宫颈癌Hela细胞miR-21基因的表达,增强细胞的凋亡,并使细胞周期阻滞于G_0/G_1期。     

miR-21对乳腺癌MCF-7细胞增殖、迁移和侵袭行为的影响

目的探讨miR-21对乳腺癌MCF-7细胞增殖、迁移和侵袭行为的影响。方法采用不同浓度的miR-21慢病毒转染乳腺癌MCF-7细胞,通过MTT实验检测乳腺癌MCF-7细胞增殖情况,采用免疫印迹与双荧光素酶实验检测miR-21与PDCD4蛋白的调控关系,使用划痕愈合实验及Transwell侵袭实验检测miR-21对乳腺癌MCF-7细胞迁移和侵袭能力的影响。结果对照组与使用浓度为0、20、40 nmol/L的miR-21-inhibitor慢病毒转染后的乳腺癌MCF-7细胞增殖速度分别为(81.24±4.65)×10`4、(72.15±6.19)×10~4、(70.25±5.59)×10~4、(52.92±7.08)×10~4,差异均统计学意义(P0.05)。抑制miR-21表达后,PDCD4蛋白的表达水平相应下调,且转染miR-21-inhibitor后,野生型PDCD4-3'非翻译区的表达活性受到一定程度抑制。抑制miR-21表达后,乳腺癌MCF-7细胞的侵袭、迁移能力均受到抑制,miR-21-inhibitor组与对照组细胞侵袭数量及细胞迁移率比较差异有统计学意义(P0.05)。结论 miR-21通过调控PDCD4蛋白的表达以影响乳腺癌MCF-7细胞的迁移和侵袭行为。     

MicroRNA-21靶向程序性细胞死亡因子4影响胃癌细胞的迁移

目的:通过调控MicroRNA-21,研究其对胃癌细胞中程序性细胞死亡因子4(PDCD4)表达及对胃癌细胞迁移的影响。方法:将MGC-803人胃癌细胞分为空白对照组,转染不相关siRNA(小干扰RNA)的阴性对照组,转染miRNA-21质粒的mir21组,转染miRNA-21抑制物的mir21 Inhibitor组,转染PDCD4siRNA的PDCD4siRNA组。分别采用实时定量PCR及Western-blot方法检测细胞中PDCD4基因及蛋白水平,Transwell小室法观察细胞迁移能力。结果:实时定量PCR及Western-blot结果显示,与空白对照组比较,mir21组PDCD4表达明显下降,48 h差异有统计学意义(P<0.05);mir21 Inhibitor组PDCD4表达量明显增多(P<0.05),转染miRNA-21抑制剂和PDCD4siRNA后,细胞迁移能力明显受到抑制,与空白对照组相比,差异有统计学意义(P<0.01)。结论:MicroRNA-21可靶向调控胃癌细胞PDCD4,抑制microRNA-21的表达后可抑制胃癌细胞的迁移能力。     

miRNA-21对宫颈癌Hela细胞顺铂化疗增敏作用的研究

目的探讨microRNA-21 (miRNA-21)对宫颈癌Hela/DDP耐药的影响及其相关机制。方法实时荧光定量PCR(Real-PCR)检测Hela细胞、Hela/DDP、转染后miR-21-siRNA组及miR-21-neg中miRNA-21的表达量。MTT法检测顺铂(DDP)对Hela/DDP增殖的影响;流式细胞仪分析Hela/DDP细胞周期及凋亡率;Western blot检测顺铂处理过的Hela/DDP中PTEN蛋白的表达。结果在Hela/DDP组中miRNA-21明显高于Hela组,差异具有统计学意义(P0.05);转染miRNA-21siRNA后,Hela/DDP中miRNA-21明显低于空白对照组及阴性对照组,差异具有统计学意义(P0.05);与空白对照组及阴性对照组相比,顺铂作用Hela/DDP后,转染miRNA-21 siRNA组细胞增殖抑制率、凋亡率及PTEN蛋白表达水平均升高,并呈现G0/G1期阻滞作用,差异具有统计学意义(P0.05)。结论 miRNA-21可抑制Hela/DDP细胞增殖、阻滞细胞周期和诱导细胞凋亡,进而提高Hela/DDP细胞对顺铂的敏感性,可能与宫颈癌Hela/DDP细胞中PTEN蛋白合成增加有关。     

miR-421靶向PDCD4促进前列腺癌DU145细胞增殖的实验研究

目的研究MicroRNA-421(miR-421)促进前列腺癌DU145细胞增殖的作用及潜在的分子机制。方法培养前列腺癌DU145细胞,分为对照组、miR-阴性对照(NC)组、miR-421组、miR-421+pcDNA组、miR-421+pcDNA-细胞程序性死亡基因4(PDCD4),miR-NC组转染miR-NC、miR-421组转染miR-421、miR-421+pcDNA组转染miR-421及pcDNA质粒、miR-421+pcDNA-PDCD4组转染miR-421及pcDNA-PDCD4质粒,MTS法测定细胞增殖活力,荧光定量PCR测定PDCD4的mRNA表达水平,western blot测定PDCD4的蛋白表达水平,双荧光素酶报告基因实验验证miR-421与PDCD4的靶向结合。结果与对照组及miR-NC组比较,miR-421组的增殖活力增加,PDCD4的表达水平及野生型PDCD4双荧光素酶报告基因的荧光活力均降低(P<0.05);与miR-421+pcDNA组比较,miR-421+pcDNAPDCD4组的增殖活力降低,PDCD4的表达水平增加(P<0.05)。结论 miR-421对前列腺癌DU145细胞的增殖具有促进作用,靶向抑制PDCD4是miR-421发挥这一作用的潜在分子机制。     

miR-21在前列腺癌中的表达及抑制其表达对前列腺癌细胞增殖凋亡的影响

目的探讨抑制前列腺癌细胞中miR-21基因表达对癌细胞增殖和凋亡的影响及机制。方法 RTPCR检测前列腺癌及癌旁组织中miR-21基因的mRNA表达;空白对照组(Control)、阴性对照组(NC)和miR-21 inhibitor组转染人前列腺癌PC-3细胞,48 h后RT-PCR检测各组细胞中miR-21基因的mRNA表达;CCK8实验检测细胞增殖;流式细胞仪检测细胞凋亡;Western blot检测ki67、Cleaved caspase3、β-catenin、Cyclin D1蛋白表达。结果前列腺癌组织中miR-21的mRNA表达显著高于癌旁组织(P0.01);转染miR-21 inhibitor后miR-21的表达显著降低;NC组细胞存活率、细胞凋亡率及ki67、Cleaved caspase3、β-catenin、Cyclin D1蛋白表达与对照组,差异无统计学意义(P0.05),miR-21 inhibitor组细胞存活率及ki67、β-catenin、Cyclin D1蛋白表达显著低于对照组,细胞凋亡率及Cleaved caspase3蛋白表达显著高于对照组(P0.01)。结论抑制前列腺癌细胞中miR-21基因的表达可降低癌细胞的增殖及诱导细胞凋亡,其机制与抑制Wnt/β-catenin信号通路有关。     

miR-124对C6胶质瘤细胞增殖和迁移能力的影响

目的:研究miR-124对C6胶质瘤细胞增殖和迁移能力的影响。方法:体外培养C6胶质瘤细胞,依据转染不同分为空白对照组、阴性对照组和miR-124拟似物组。实时荧光PCR检测转染效率,绘制生长曲线,MTT实验检测miR-124对C6细胞增殖能力影响,计算抑制率。划痕实验检测miR-1 2 4对C 6细胞迁移能力影响,计算迁移率。结果:生长曲线显示miR-124拟似物组C 6细胞生长能力受到明显抑制,转染后第3天,miR-124拟似物组C6细胞数目显著低于阴性对照组(5.410±0.463v s.6.917±0.385;P0.01);miR-1 2 4拟似物组mRNA表达量显著高于阴性对照组和空白对照组(5.92±0.56 vs.0.93±0.13和1.00±0.12;P0.01);MTT增殖实验显示miR-124抑制C6细胞增殖能力,miR-124拟似物组抑制率显著高于阴性对照物组(42.90±5.169 vs.10.24±3.351;P0.01);划痕实验显示miR-124明显抑制C6细胞迁移能力,阴性对照组迁移率显著高于miR-124拟似物组(98.79±1.210vs.81.72±5.972;P0.05)。结论:miR-124能够显著抑制C6胶质瘤细胞的增殖和迁移能力。     

miR-103a-3p通过调控FEZF1/CDC25A途径影响肝癌细胞的增殖和凋亡

目的探究miR-103a-3p在肝癌细胞的表达及其与锌指蛋白1(FEZF1)/细胞分裂周期25A蛋白(CDC25A)途径的关系。方法体外培养人肝癌Hep G2、BEL-7402细胞,将转染试剂、miR-103a-3p阴性对照质粒、miR-103a-3p mimis质粒分别转染细胞,命名为空白对照组、miR-103a-3p阴性转染组、miR-103a-3p过表达组。实时定量PCR(qRT-PCR)检测miR-103a-3p mRNMA表达;MTT法检测细胞增殖情况;平板细胞克隆形成试验检测菌落形成情况;流式细胞仪检测细胞凋亡以及周期分布情况;蛋白免疫印迹法(WB)检测细胞中FEZF1、CDC25A蛋白表达情况;荧光素酶活性检测miR-103a-3p与FEZF1靶向调控关系。结果在Hep G2细胞、BEL-7402细胞中,与空白对照组、阴性转染组相比,miR-103a-3p过表达组miR-103a-3p mRNA表达显著升高(P0.05)。随着细胞培养时间延长,miR-103a-3p过表达组细胞抑制率逐渐升高,在同一时间点,与空白对照组、阴性转染组相比,miR-103a-3p过表达组细胞抑制率均升高(P0.05)。与空白对照组、阴性转染组相比,细胞菌落数量显著降低,细胞凋亡率显著升高,G0/G1期DNA含量升高,S期细胞DNA含量下降,FEZF1、CDC25A蛋白表达均降低(P0.05)。荧光素酶活性测定显示,在3'UTR-Wt的细胞中,与阴性转染组相比,miR-103a-3p过表达组荧光素酶活性显著降低(P0.05);而3'UTR-Mut的细胞中,阴性转染组与miR-103a-3p过表达组荧光素酶活性无显著差异(P0.05)。结论 miR-103a-3p过表达后能够抑制肝癌细胞的增殖,引起细胞周期G0/G1期阻滞,促进其凋亡,其机制可能与靶向调控FEZF1/CDC25A表达有关。     

破骨细胞外泌体miR-183-5p靶向程序性细胞死亡因子4促进肺腺癌细胞增殖及侵袭

目的 探讨破骨细胞来源外泌体(osteoclast-derived exosomes)中miR-183-5p靶向程序性细胞死亡因子4(programmed cell death 4, PDCD4)对肺腺癌侵袭增殖的影响。方法 利用RAW264.7细胞诱导分化破骨细胞并提取外泌体,透射电镜及粒度仪对其进行鉴定。将外泌体与A549细胞共培养,通过CCK8法检测细胞活性,观察miR-183-5p对A549细胞增殖的作用;检测A549细胞周期分布,观察miR-183-5p对细胞周期的作用;利用Transwell侵袭实验,观察miR-183-5p对A549细胞侵袭能力的作用。Western blotting检测PDCD4、周期蛋白D1(Cyclin D1, CCND1)和周期蛋白依赖性激酶4(cyclin dependent kinase, CDK4)相对表达,观察miR-183-5p靶蛋白表达情况及其对A549细胞侵袭、增殖的影响。结果 外泌体呈边缘透亮清晰的椭圆形囊泡状结构,粒径分布范围30-200nm左右,荧光探针发现外泌体能够较快的被肺腺癌细胞A549摄取,并且共培养后A549细胞中miR-183-5p表达水平明显增高(p<0.01)。miR-183-5p高表达可以明显促进A549细胞的侵袭、细胞增殖和周期(p<0.05)。A549细胞中miR-183-5p过表达后,PDCD4表达降低,同时CCND1、CDK4也高表达(p<0.05)。结论 破骨细胞来源的外泌体与A549细胞共培养可提高细胞中miR-183-5p的表达,miR-183-5p通过靶向抑制PDCD4表达可促进肺腺癌细胞侵袭、增殖和迁移能力。     

Retracted Article: Linc00472 suppresses breast cancer progression and enhances doxorubicin sensitivity through regulation of miR-141 and programmed cell death 4

The existence of drug resistance strikingly hampers the therapy of many malignancies, including breast cancer. Long non-coding RNAs (LncRNAs) have been reported to participate in the regulation of various biological processes associated with cancer progression. Whereas, the role of linc00472 in breast cancer pathogenesis and doxorubicin (ADR) resistance have not been well elucidated. In the present study, it is found that linc00472 expression was decreased in breast cancer tissues and cells. Moreover, higher linc00472 expression was positively associated with favorable disease status and prognosis for breast cancer patients. Functional analyses revealed that linc00472 overexpression suppressed proliferation and invasion, facilitated apoptosis and enhanced ADR sensitivity in breast cancer cells. Mechanistic studies discovered that linc00472 acted as a competing endogenous RNA (ceRNA) of miR-141 to sequester miR-141 from its target mRNA PDCD4 (programmed cell death 4). Furthermore, the inhibition effect of linc00472 on breast cancer cell progression and ADR resistance could be partly abrogated by miR-141 up-regulation or PDCD4 knockdown. In vivo assays also demonstrated that linc00472 hindered tumor growth by suppressing miR-141 expression and enhancing PDCD4 expression. In conclusion, linc00472 blocked breast cancer progression and induced ADR sensitivity through regulation of miR-141 and PDCD4, highlighting a potential therapeutic strategy for breast cancer patients.

Linc00472 expression was down-regulated in breast cancer tissues and cells, and was associated with the development and prognosis of breast cancer.     

MicroRNA-21在弥漫大B细胞淋巴瘤中的表达及其意义

本研究旨在检测弥漫大B细胞淋巴瘤（diffuse large B cell lymphoma,DLBCL）细胞株中miR-21（micro RNA-21）的表达以及调控PDCD4及PTEN 2种基因对肿瘤生长、浸润及转移能力的影响,并探讨以miR-21作为靶分子对DLBCL基因治疗的可能性。首先,使用荧光定量PCR（qRT-PCR）检测3种DLBCL细胞株中miR-21的表达水平;利用转染anti-miR-21来降低miR-21的表达量,研究miR-21对DLBCL的生物行为可能产生的作用;通过荧光素酶报告基因检测,定量RT-PCR和Western blot来验证miR-21与PDCD4和PTEN的关系。结果表明：ABC-型DLBCL细胞株中miR-21的表达明显高于GCB-型DLBCL;RNA反义抑制技术特异性降低OCI-LY3和OCILY10细胞miR-21表达后,细胞增殖、侵袭能力受到明显抑制,凋亡增加;荧光素酶报告、定量RT-PCR和Western blot检测表明,PDCD4及PTEN是miR-21靶基因。结论：miR-21可能涉及DLBCL的发病机制,可能为一种新的DLBCL治疗靶分子。     

Characterization of programmed cell death 4 in multiple human cancers reveals a novel enhancer of drug sensitivity

Programmed cell death 4 (Pdcd4), originally identified as an inhibitor of murine cellular transformation, inhibits protein synthesis by directly interacting with eukaryotic initiation factor 4A (eIF4A) of the translation initiation complex. The relevance of Pdcd4 to a broad range of human cancers derived from multiple tissue sites is unknown. Protein expression patterns from the National Cancer Institute drug-screening panel of 60 human cancer cells (NCI60) were analyzed by Western blot methods and revealed frequent reduction of Pdcd4 protein levels in renal-, lung-, and glia-derived tumors. Greater than mean Pdcd4 protein levels correlated with the antitumor activity of geldanamycin and tamoxifen. Stable expression of antisense PDCD4 significantly reduced the sensitivity of MCF-7 breast cancer cells to geldanamycin and to tamoxifen. Sensitivity to geldanamycin significantly increased in UO-31 renal cancer cells expressing sense PDCD4 cDNA. Increased geldanamycin sensitivity was accompanied by enhanced cell cycle arrest and apoptosis. One primary mode of inactivation of Pdcd4 in human cancers appears to involve down-regulated expression, and this down-regulation causes a decreased sensitivity to geldanamycin cytotoxicity. Thus, up-regulating Pdcd4 expression may be promising for geldanamycin-based combination therapy.     

Retracted Article: Exosome-derived PTENP1 suppresses cisplatin resistance of bladder cancer (BC) by suppressing cell proliferation,migration and inducing apoptosis via the miR-103a/PDCD4 axis

Bladder cancer (BC) is a lethal cancer that threatens the health of millions of people. Chemotherapy drug resistance, for example, cisplatin (DDP) resistance, is a huge limitation for BC therapy. PTEN pseudogene-1 (PTENP1) has been identified as a significant biomarker of multiple cancers. Therefore, it is essential to illuminate the molecular mechanism of PTENP1 in BC cell DDP resistance and progression. Serum exosomes were isolated using an ExoQuick precipitation kit. Serum exosomes were round-shaped vesicles of 100 ± 60 nm in size. The expression of PTENP1 was down-regulated in serum exosomes isolated from cisplatin non-responsive patients compared with responsive patients. ROC curves certified the diagnostic value of PTENP1. Apparently, PTENP1 transfection inhibited DDP-resistant BC cell proliferation, migration, cisplatin resistance and facilitated apoptosis. Next, we discovered that PTENP1 was a sponge of miR-103a, while PDCD4 was a target of miR-103a. More importantly, PTENP1 regulated DDP-resistant cell viability, migration, apoptosis and cisplatin resistance by interacting with the miR-103a/PDCD4 axis. In addition, PTENP1 hindered tumor growth of cisplatin-resistant mice. Exosome-derived PTENP1 suppressed the DDP resistance of BC by inhibiting cell proliferation, migration and promoting apoptosis through regulating the miR-103a/PDCD4 axis, representing a targeted therapy for DDP-resistant BC patients.

Bladder cancer (BC) is a lethal cancer that threatens the health of millions of people.     

缺血性脑卒中患者血清miR-155和PDCD4mRNA表达水平与颈动脉支架成形术后再狭窄的相关性研究

目的　了解缺血性脑卒中（cerebral ischemic stroke,CIS）患者血清微小RNA-155（microRNA-155,miR-155）、程序性细胞死亡因子4（programmed cell death factor 4,PDCD4）信使RNA（messenger RNA,mRNA）表达水平与颈动脉支架成形术（carotid artery stenting,CAS）后再狭窄的相关性。方法　选择2016 年2 月～ 2020 年7 月在凉山彝族自治州第二人民医院确诊的CIS 患者120 例作为CIS 组,同期选取健康体检者125 例为对照组。实时荧光定量PCR 法测定CIS 组术前和术后24 h,对照组体检时血清miR-155 和PDCD4 mRNA 表达水平;术后180 天对CIS 患者进行脑多普勒超声检查和颈动脉彩超检查,观察CIS 患者颈动脉狭窄程度、不稳定斑块数量、斑块长度及斑块厚度,测定患者血浆粘度、全血黏度、红细胞比容和纤维蛋白原水平;采用受试者工作特征（receiver operating characteristic,ROC）曲线分析血清miR-155 和PDCD4 mRNA 表达水平对CIS 患者行CAS 术后再狭窄的诊断价值。结果　与对照组相比,CIS 组术前血清miR-155 表达水平显著降低（0.31±0.06 vs 1.01±0.20）（q=45.189,P=0.000）,CIS 组术前（2.23±0.45 vs 1.03±0.20）和术后24 h（1.19±0.29 vs 1.03±0.20）血清PDCD4 mRNA 表达水平均显著升高（q=40.422,5.390,均P=0.000）,差异均有统计学意义;与CIS 组术前相比,CIS 组术后24 h 血清miR-155 表达水平显著升高（0.97±0.21 vs 0.31±0.06）（q=42.179,P=0.000）,血清PDCD4 mRNA 表达水平显著降低（1.19±0.23 vs2.23±0.45）（q=34.680,P=0.000）,差异均有统计学意义;与未狭窄组相比,再狭窄组颈动脉狭窄程度（21.31±5.33vs 9.11±2.52）、术后24 h 血清PDCD4 mRNA 表达水平（2.18±0.54 vs 1.06±0.26）、不稳定斑块数量（5.51±1.27 vs2.87±0.71）、斑块长度（7.60±1.90mm vs 3.16±0.79mm）、斑块厚度（4.60±1.15mm vs 2.84±0.71mm）、血浆黏度（15.03±3.85mPa·s vs 11.01±2.75mPa·s）、全血粘度(5.84±1.46mPa·s vs 3.71±0.92mPa·s）、红细胞比容（58.98%±17.74%vs 46.51%±11.62%）和纤维蛋白原（4.93±1.23g/L vs 3.01±0.75g/L）显著升高,差异均有统计学意义（t=4.277 ～ 17.874,均P<0.05）,术后24 h 血清miR-155 表达水平显著降低（0.36±0.09 vs 1.03±0.25）,差异有统计学意义（t=9.074,P=0.000）。ROC曲线结果表明,术后24 h血清miR-155和PDCD4 mRNA表达水平诊断CAS术后再狭窄的AUC分别为0.778（95% CI 为0.692~0.864）和0.838（95% CI 为0.749~0.926）。结论　血清miR-155 和PDCD4 mRNA 的异常表达与CIS病情及CAS 术后再狭窄联系密切,对CAS 术后再狭窄具有一定的诊断价值。     

Cisplatin upregulates MSH2 expression by reducing miR-21 to inhibit A549 cell growth

miR-21 can act as an oncogene. MSH2 has been reported that it involved in the DNA mismatch repair (MMR) system and overexpression of MSH2 can induce cell apoptosis. We predicted that MSH2-3′-untranslated region (3′-UTR) was targeted by miR-21 using microRNA analysis softwares. To further explore the roles of miR-21 and MSH2 in A549 cells, we constructed pcDNA-GFP-msh-UTR vector (including MSH2-3′-UTR) to transfect A549 cells with miR-21, GFP positive cells were estimated under a fluorescence microscopy and by flow cytometry. We found miR-21 could obviously downregulate the expression of MSH2, which was further proved by western blotting. Moreover, we treated A549 cells with cisplatin and found that cisplatin could inhibit A549 cell growth in vitro and in vivo. We also found that cisplatin could downregulate miR-21 expression, while increase MSH2 expression in A549 cells. Our results demonstrated that cisplatin could upregulate the expression of MSH2 through downregulating miR-21 to inhibit A549 cell proliferation, which provides new gene targets for drug design or cancer therary.