微小RNA-101对胰腺癌PANC1细胞增殖、凋亡和侵袭的影响及其作用机制

目的探讨微小RNA-101(miR-101)对胰腺癌PANC1细胞增殖、凋亡和侵袭的影响及其可能的作用机制。方法取对数生长期PANC1细胞分为5组, 除对照组外, 分别为转染miRNA模拟体的阴性对照(mimic-NC)组、转染miRNA-101模拟体(miR-101 mimic)组、转染miRNA抑制剂的阴性对照(inhibitor-NC)组、转染miRNA-101抑制剂(miR-101 inhibitor)组。采用qRT-PCR检测miR-101相对表达水平, MTT法检测细胞增殖率, 流式细胞仪检测细胞凋亡率, Transwell实验检测细胞侵袭能力, qRT-PCR和蛋白质免疫印迹法检测IL-6、JAK2、p-JAK2、STAT3、p-STAT3 mRNA蛋白的相对表达水平, 双荧光素酶报告基因法检测miR-101与IL-6的靶向关系。结果转染48 h后, 对照组、mimic-NC组、miR-101 mimic组、inhibitor-NC组、miR-101 inhibitor组PANC1细胞miR101表达水平分别为1.98±0.12、2.01±0.18、6.73±0.23、2...     

HMGN5基因与子宫内膜癌临床病理特征及生物学行为的关系

目的 研究高迁移率族核小体结合域(HMGN)5与子宫内膜癌患者临床病理特征相关性及对人子宫内膜癌细胞HEC-1A细胞增殖、侵袭和转移的影响。方法 连续纳入2020年1月至2021年6月柳州市人民医院实施根治性切除术的子宫内膜癌病例79例。应用反转录聚合酶链式反应(RT-PCR)法对子宫内膜癌肿瘤组织和癌旁正常组织的HMGN5 mRNA表达量进行检测。HEC-1A细胞复苏后培养,取对数期细胞随机将其分为HMGN5促进组(转染NMGN5干扰序列)、HMGN5正常组(转染HMGN5干扰对照序列)和空白对照组(不做处理),进行转染48 h并进行后期实验。应用RT-PCR法对HEC-1A细胞中HMGN5 mRNA表达进行检测;检测3组细胞HEC-1A细胞增殖活力;应用划痕实验对HEC-1A细胞迁移能力进行检测;应用Transwell小室检测细胞侵袭能力。结果 子宫内膜癌组HMGN5 mRNA相对表达量显著高于癌旁组织(P<0.05);有淋巴结转移、中低分化及FIGO分期Ⅱ～Ⅳ期患者的HMGN5的相对表达量要明显低于无淋巴结转移、高分化及FIGO分期Ⅰ期患者(P<0.05);转染后,与...     

miRNA-21对结直肠癌耐药细胞ATM/CHK2/CDC25A信号通路的影响

目的 探讨miRNA-21在结直肠癌奥沙利铂(L-OHP)耐药细胞株HCT116/L-OHP中对毛细血管扩张性共济失调突变基因(ATM)/细胞周期检测点激酶(CHK)2/细胞分裂周期素(CDC)25A信号通路的影响。方法 应用细胞转染技术干扰HCT116/L-OHP细胞中miRNA-21的表达，采用实时荧光定量PCR(RT-qPCR)检测HCT116/L-OHP中ATM、CHK2、CDC25A和细胞周期蛋白依赖性激酶(CDK)2 mRNA表达，Western印迹检测该细胞株中miRNA-21表达变化后ATM、CHK2、CDC25A和CDK2蛋白表达。结果 转染miRNA-21 mimic使miRNA-21表达升高时，ATM、CHK2、CDC25A和CDK2 mRNA和蛋白的表达均随之升高，且与未转染组和阴性转染组相比较差异均具有统计学意义(P<0.05);转染miRNA-21 inhibitor使miRNA-21表达降低时，ATM、CHK2、CDC25A和CDK2 mRNA和蛋白的表达均随之降低，且与未转染组和阴性转染组相比较差异均具有统计学意义(P<0.05)。结论 在结...     

微小RNA-139-5p在缺氧诱导的原代心肌细胞凋亡中的作用及机制

目的研究微小RNA(miR)-139-5p在缺氧诱导的心肌细胞凋亡中的作用及机制。方法分离培养新生Wistar大鼠原代心肌细胞,将细胞进行无特殊(对照)、miR-139-5p模拟物(mimic)转染、miR-139-5p抑制剂(inhbitor)转染、缺氧、转染mimic+缺氧处理、转染inibitor+缺氧处理,转染mimic后采用逆转录-聚合酶链反应(RT-PCR)检测心肌细胞中miR-139-5p的表达;采用流式细胞术检测对照、转染、缺氧、转染mimic+缺氧、转染inibitor+缺氧处理后细胞的凋亡率;Western blot检测心肌细胞wnt信号通路wnt1、β-连环蛋白(β-catenin)以及B细胞淋巴瘤因子2(bcl-2)的蛋白表达水平。结果与对照无特殊处理细胞比较,转染miR-139-5p mimic后miR-139-5p的相对表达水平明显增高(14.01±0.50比1.51±0.16,P0.05)。与对照和转染处理后的细胞相比,缺氧后细胞的wnt1、β-catenin、bcl-2蛋白表达升高(均P0.05);缺氧后细胞凋亡率明显升高[(20.70±0.40)%比(0.88±0.18)%、(1.00±0.14)%,P0.05]。与缺氧后的细胞相比,转染mimic+缺氧细胞的wnt1、β-catenin、bcl-2蛋白表达明显降低(均P0.05),转染inhibitor+缺氧细胞的wnt1、β-catenin、bcl-2蛋白表达明显升高(均P0.05),转染mimic+缺氧细胞凋亡率明显升高[(36.10±2.30)%比(20.70±0.40)%,P0.05],转染inhibitor+缺氧细胞凋亡率明显降低[(14.5±1.10)%比(20.70±0.40)%,P0.05]。对照和转染处理后细胞的凋亡率和蛋白表达差异无统计学意义(P0.05)。转染mimic和缺氧对wnt1(F=53.887,P_(交互)0.05)、β-catenin(F=19.112,P_(交互)0.05)、bcl-2(F=15.368,P_(交互)=0.001)表达的影响存在交互作用。结论 miR-139-5p通过抑制wnt/β-catenin信号通路,降低心脏的保护因子bcl-2的表达,增加缺氧状态下心肌细胞的凋亡。     

miRNA-638对巨噬细胞内胆固醇外流的影响及其发生机制研究

背景流行病学研究表明生活方式、饮食结构等外部因素的改变与遗传因素相互作用导致人体脂代谢紊乱,而胆固醇逆转运过程能够维持体内胆固醇稳态。目的探讨miRNA-638对巨噬细胞内胆固醇外流的影响及其发生机制。方法实验于2016年9月-2017年9月完成。采用100 ng/ml佛波酯诱导人髓系白血病单核细胞(THP-1)贴壁并分化为巨噬细胞,根据预实验结果选取浓度为20 nmol/L的miRNA-638模拟物及其阴性对照物进行转染。采用NBD-胆固醇分别检测转染miRNA-638模拟物及其阴性对照物巨噬细胞经载脂蛋白AI(ApoAI)、高密度脂蛋白(HDL)及标准血清外流液介导的胆固醇外流率;采用实时荧光定量PCR及Western blotting检测转染miRNA-638模拟物及其阴性对照物巨噬细胞中三磷酸腺苷结合盒转运体A1(ABCA1)、三磷酸腺苷结合盒转运体G1(ABCG1)和极低密度脂蛋白受体(VLDLR)mRNA及其相对表达量。结果转染miRNA-638模拟物巨噬细胞经ApoAI外流液介导的NBD-胆固醇外流率低于转染阴性对照物巨噬细胞(P<0.05);转染miRNA-638模拟物巨噬细胞与转染阴性对照物巨噬细胞经HDL、标准血清外流液介导的NBD-胆固醇外流率比较,差异无统计学意义(P>0.05)。转染miRNA-638模拟物巨噬细胞的ABCA1、ABCG1、VLDLR mRNA相对表达量低于转染阴性对照物巨噬细胞(P<0.05)。转染miRNA-638模拟物巨噬细胞中ABCA1、ABCG1相对表达量低于转染阴性对照物巨噬细胞(P<0.05);转染miRNA-638模拟物巨噬细胞与转染阴性对照物巨噬细胞中VLDLR相对表达量比较,差异无统计意义(P>0.05)。结论miRNA-638可减少由ApoAI介导的巨噬细胞内胆固醇外流,该作用可能是通过调控ABCA1/ABCG1的表达实现的。     

微小RNA-126在食管癌组织中的表达及其对食管癌EC109细胞增殖和迁移的影响

目的探讨微小RNA(mi RNA)-126在食管癌组织中的表达及其对食管癌EC109细胞增殖和迁移的影响。方法收集该院胸外科手术切除且保存完整的食管癌及相应癌旁组织标本30例,通过实时荧光定量PCR(RT-PCR)检测癌组织和癌旁组织中mi RNA-126 m RNA表达;mi RNA-126 mimi和control转染EC109细胞,设置阴性对照组(转染mi RNA mimic control)、转染组、空白对照组(仅转染试剂),PCR检测转染EC109细胞mi RNA-126表达,转染细胞培养1、3、5 d后MTT检测细胞增殖情况,Transwell小室法检测细胞迁移活力。结果癌组织中mi R-126 m RNA低于癌旁组织(P0.05);转染EC109细胞后mi RNA-126在阴性对照组和空白对照组表达量均低于转染组(P0.05);三组转染EC109细胞随着培养时间延长,细胞OD值均逐渐升高(P0.05),培养第3、5天细胞OD值组间差异显著(P0.05),其中转染组EC109细胞OD值均低于阴性对照组和空白对照组(P0.05);转染EC109细胞迁移活力检测结果显示,转染组OD值低于阴性对照组和空白对照组(P0.05)。结论食管癌组织中mi RNA-126低表达,并且对食管癌EC109细胞增殖和迁移具有一定抑制作用。     

miR-9-3p对细胞脂代谢调节功能及可能机制

目的 检测microRNA-9-3p (miR-9-3p)对HepG2细胞增殖及脂质代谢的影响,探讨其对细胞沉默信息调节因子(Sirt-1)蛋白表达的影响。方法 以HepG2细胞为体外细胞模型,应用体外细胞培养技术、噻唑蓝(MTT)比色法、转染技术和Western印迹,以miR-9-3p mimic、miR-9-3p inhibitor转染为干预手段,应用MTT比色法、Western印迹检测正常对照组、miR-9-3p mimic组、anti-miR-9-3p组3组HepG2细胞增殖水平、总胆固醇(TC)及三酰甘油(TG)表达水平及Sirt-1蛋白的表达。结果 miR-9-3p mimic组miR-9-3p表达水平显著高于正常对照组(P<0.05);anti-miR-9-3p组miR-9-3p表达水平显著低于正常对照组(P<0.05);与正常对照组相比,miR-9-3p mimic组HepG2细胞增殖显著增加(P<0.05);anti-miR-9-3p组HepG2细胞增殖显著抑制(P<0.05);miR-9-3p mimic组HepG2细胞TC及TG表达水平显著...     

miRNA-200a调控淋巴瘤细胞增殖迁移的机制

目的探讨微小RNA-200a(miRNA-200a)对淋巴瘤DOHH-2细胞增殖的影响及其机制。方法在淋巴瘤细胞DOHH-2中转染miRNA-200a,采用qRT-PCR和Western印迹的方法检测DOHH-2细胞进行转染的24 h后的miRNA-200a和PTEN mRNA和蛋白的表达情况,采用CCK8检测各组细胞增殖,采用划痕实验检测各组细胞迁移细胞速率,每组重复6次。结果转染miRNA-200a后,与对照组或阴性对照组比,miRNA-200a转染组的miRNA-200a水平显著增高(P0.05),PTEN水平显著降低(P0.05),对照组和阴性对照组的miRNA-200a及PTEN mRNA及蛋白表达差异无统计学意义(P0.05),转染miRNA-200a后,与对照组或阴性对照组比,miRNA-200a转染组的相对细胞活力和细胞迁移速率明显增高(P0.05)。结论 miRNA-200a调控淋巴瘤细胞DOHH-2的增殖与迁移,其作用机制与miRNA-200a反向调控肿瘤抑制因子PTEN有关。     

17β-E2对子宫内膜癌细胞增殖及其p-Erk表达的影响

目的探讨17β-雌二醇（E2）对人子宫内膜腺癌Ishikawa和HEC-1A细胞增殖、周期及其p-Erk表达的影响。方法用MTT法、流式细胞术检测不同浓度17β-E2作用Ishikawa和HEC-1A不同时间后的细胞吸光度值及细胞周期,免疫细胞化学法检测细胞的p-Erk。结果随17β-E2浓度增加,Ishikawa细胞吸光度值上升,并呈时间依赖性（P〈0.01）,G0-G1期细胞比例下降（P〈0.01）,S期比例升高（P〈0.01）;HEC-1A细胞吸光度值及细胞周期无明显变化（P均〉0.05）。10-6mol/L的17β-E2作用于Ishikawa细胞30 min、作用于HEC-1A细胞15 min时,其p-Erk表达最强。结论17β-E2可以促进Ishikawa细胞的增殖和周期进展,而且可以通过非转录效应迅速激活Ishikawa和HEC-1A中MAPK信号转导通路。     

miR21介导Ang(1-7)对肺成纤维细胞AngⅡ诱导的NLR3炎性体激活的抑制作用

目的探讨miR21对肺成纤维细胞增殖和凋亡的影响,及其可能的作用机制。 方法将miR21 NC、inhibitor和mimic转染至原代肺成纤维细胞,按照细胞处理方式分为空白对照组、miR21 NC组、inhibitor组和mimic组。利用CCK-8细胞增殖法检测miR21对肺成纤维细胞增殖的影响;流式细胞仪检测miR21对肺成纤维细胞凋亡的影响;通过RT-PCR法检测转染前后肺成纤维细胞miR21、Ang(1-7)基因水平的表达;应用免疫印迹Western blot检测转染前后miR21、NLRP3蛋白表达水平变化。 结果CCK8及流式细胞仪检测显示,miR21具有显著促进肺成纤维细胞增殖、抑制肺成纤维细胞凋亡的作用;RT-PCR分析显示,细胞转染成功后,miR21 mimic/inhibitor组分别抑制/促进了Ang(1-7)表达,促进/抑制了AngⅡ表达(P<0.05),即miR21具有抑制Ang(1-7),促进AngⅡ表达的效果。Western Blot分析表明,miR21 mimic/inhibitor具有上调/下调AngⅡ、NLRP3蛋白表达水平的作用,与NC对照组相比,差异具有统计学意义(P<0.05)。 结论miR21具有促进肺成纤维细胞增殖、抑制肺成纤维细胞凋亡的作用,其机制可能与miR21介导Ang(1-7)对肺成纤维细胞AngⅡ诱导的NLR3炎性体激活相关。     

MiR-145 participates in the pathogenesis of aortic dissection by promoting smooth muscle remodeling

Background Ascending aortic dissection(AD)is a lethal vascular disease with a high mortality. Smooth muscle cell is an important component of aortic medial wall and therefore its role in the pathogenesis of aortic dissection has been emphasized. Upregulation of micro RNA-145 has been reported to been involved in vessel remodeling. The present study aimed to investigate the role of mi RNA-145 in aortic medial wall remodeling and the pathogenesis of AD. Methods Between August 2014 and November 2016,58 patients with aortic dissection and 58 healthy controls were enrolled in the study. Serum levels of mi RNA-145 had been measured by q RTPCR. In in vitro studies,vascular smooth muscular cells were transfected with mi RNA-145 mimic or negative control,respectively. The expressions of osteopontin and type III collagen were measured by western blot. Results Serum levels of mi RNA-145 was significantly higher in patients with aortic dissection as compared with healthy controls(P 0.001). Furthermore,the expression of osteopontin and type III collagen increased significantly in vascular smooth muscular cells transfected with mi RNA-145 mimic than negative controls. Conclusion Mi RNA-145 could be participating in the the pathogenesis of aortic dissection through influencing subtype transformation of smooth vascular cells and expression of collagen,indicating a novel target for intervention of aortic dissection.     

微小RNA-146a转染的H9c2心肌细胞高糖培养下核因子-κcBp65和Ⅰ型胶原的表达

目的 观察微小RNA（miRNA）-146a转染H9c2心肌细胞高糖培养下NF-κBp65和Ι型胶原表达。 方法 高糖培养H9c2细胞系,分为增强组（EC）、阴性对照组（NC）、空白对照组（BC）。采用Lipofectamine 2000 Transfection Reagent分别转染miRNA-146a增敏剂（50 nmol/L）、错义链（50 nmol/L）、PBS,于48 h后Real-time PCR测定miRNA-146a水平,Western-blot测定NF-κBp65和Ⅰ型胶原水平。 结果 转染后,EC组H9c2细胞miRNA-146a水平高于NC组和BC组（P〈0.05）,NF-κBp65和Ⅰ型胶原蛋白表达量低于NC组和BC组（P〈0.05）。 结论 miRNA-146a参与了高糖培养H9c2心肌细胞纤维化,其机制与NF-κBp65和Ι型胶原表达有关。     

miRNA-448 inhibits cell growth by targeting BCL-2 in hepatocellular carcinoma

BackgroundIncreasing evidence indicates that aberrant micro (mi)RNA-448 expression plays a critical role in the progression of several human cancers. However, the function of miRNA-448 in hepatocellular carcinoma (HCC) has not been fully investigated.MethodsmiRNA-448 expression levels in HCC tissues, adjacent non-cancerous tissues (ANTs), and HCC cell lines were examined by quantitative real-time polymerase chain reaction (qRT-PCR). HCC cells were treated with a miRNA-448 mimic or inhibitor, followed by cell viability measurements with the CCK-8 assay. Venn diagram analysis predicted, and dual luciferase reporter assays verified, the target gene of miRNA-448. Expression of the target gene was detected by qRT-PCR and immunohistochemistry. Growth of miRNA-448- or target gene-expressing HCC xenograft tumors in nude mice was measured.ResultsmiRNA-448 was expressed at a lower level in HCC tissues than ANTs, and correlated with a larger tumor size, incomplete tumor encapsulation, and advanced Barcelona Clinic Liver Cancer stage. miRNA-448 inhibited HCC cell growth. The downstream target of miRNA-448 was BCL-2, which was highly expressed in HCC tissues and its mRNA level was negatively correlated with miRNA-448 expression. In vivo, BCL-2 attenuated the tumor inhibiting effect of miRNA-448.ConclusionmiRNA-448 functions as a tumor suppressor by targeting BCL-2 in HCC.     

Low <Emphasis Type="Italic">LATS2</Emphasis> mRNA level can predict favorable response to epirubicin plus cyclophosphamide,but not to docetaxel,in breast cancers

Purpose Putative tumor suppressor genes LATS1 and LATS2 are implicated in the regulation of the cell cycle at the G2/M and G1/S phase, respectively. This study investigated possible correlations of intra-tumoral LATS1 and LATS2 mRNA levels with response to epirubicin plus cyclophosphamide (EC) or docetaxel (DOC) treatment. Methods mRNA expression levels of LATS1 and LATS2 were determined by means of real-time PCR assay in 56 locally advanced breast cancers and 15 recurrent breast cancers treated with EC (n = 32) or DOC (n = 39). Results Among the patients treated with EC, LATS2 mRNA levels of responders (0.72 ± 0.11, mean ± SE) were significantly (P < 0.05) lower than those of non-responders (1.62 ± 0.44), and responders showed a tendency (P = 0.05) towards reduced LATS1 mRNA levels. Patients with low LATS2 mRNA levels (n = 16) showed a significantly (P < 0.05) higher response rate (75%) to EC treatment than those with high LATS2 mRNA levels (n = 16; response rate = 31%). Positive predictive value, negative predictive value, and diagnostic accuracy of LATS2 mRNA levels for prediction of response to EC were 75, 69, and 72%, respectively. On the other hand, neither LATS1 nor LATS2 mRNA levels were associated with response to DOC treatment. Conclusion These results suggest the possibility that intra-tumoral LATS2 mRNA levels may be clinically useful for the prediction of response to EC treatment by breast cancer patients. We speculate that disruption of the checkpoint function at the G1/S phase induced by down-regulation of LATS2 plays some part in the favorable response to EC.     

Construction of pETNF-P16 plasmid and its expression properties in EC9706 cell line induced by X-ray irradiation

AIM: Recombined plasmid pETNF-P16 was constructed to investigate its expression properties in esophageal squamous carcinoma cell line EC9706 induced by X-ray irradiation and the feasibility of gene-radiotherapy for esophageal carcinoma. METHODS: Recombined plasmid pETNF-P16 was constructed and transfected into EC9706 cells with lipofectamine. ELISA,Western blot, and immunocytochemistry were performedto determine the expression properties of pETNF-P16 in EC9706 after transfection induced by X-ray irradiation. RESULTS: Eukaryotic expression vector pETNF-P16 was successfully constructed and transfected into EC9706 cells. TNFα expressions were significantly increased in the transfected cells after different doses of X-ray irradiation than in those after 0Gy irradiation (1 192.330-2 026.518 pg/mL,P&lt;0.05-0.01), and the TNFα expressions and P16 were significantly higher 6-48 h after 2 Gy X-ray irradiation (358.963-585.571 pg/mL, P&lt;0.05-0.001). No P16 expression was detected in normal EC9706 cells. However, there was strong expression in the transfected and irradiation groups. CONCLUSION: X-ray irradiation induction could significantly enhance TNFα and P16 expression in EC9706 cells transfected with pETNF-P16 plasmid. These results may provide important experimental data and therapeutic potential for gene-radiotherapy of esophageal carcinoma.     

微RNA-21-5p靶向调控程序性死亡蛋白4对系统性红斑狼疮患者外周血B细胞增殖与凋亡的影响

目的 探讨微RNA(RNA-21-5p)通过靶向程序性死亡蛋白4(PDCD4)对SLE患者外周血B细胞增殖与凋亡的调控作用.方法 选择我院经临床确诊的SLE患者30例,采用梯度离心法提取外周血淋巴细胞(PBL),磁珠法分选B细胞后流式细胞术检测外周血B细胞比例;采用电转染法将细胞分为空白对照组、miRNA-21-5p阴性对照组、miRNA-21-5p Inhibitor组、PDCD4阴性对照组、PDCD4 siRNA组.于转染后48 h收集细胞.采用实时荧光定量(RT)-PCR法检测各组细胞miRNA-21-5p及PDCD4 mRNA的表达水平;采用蛋白质印迹法检测各组细胞PDCD4的蛋白表达;采用双荧光素酶靶标实验验证miR-21-5p和PDCD4的靶向关系;采用流式细胞仪检测各组细胞凋亡情况、采用CCK-8法检测各组细胞的增殖情况;采用蛋白质印迹法和RT-PCR法分别检测各组细胞Fas、FasL、CD40和CD40L的表达水平.采用独立样本t检验对2组间数据进行比较;采用单因素方差分析对多样本实验结果进行统计分析;抗dsDNA抗体阳性率比较采用χ^2检验.结果 SLE患者血清补体[C3 (0.85±0.11)g/L、C4 (0.54±0.09)g/L]较健康对照组[C3(1.16±0.17) g/L、C4(1.57±0.09) g/L]降低(t=7.524,P<0.05;t=38.471,P<0.05),而抗dsDNA抗体(47%)、IgG(15.46±0.75) g/L、IgA (2.68±0.20) g/L较健康对照组抗dsDNA抗体(17%)、IgG(11.95±0.80) g/L、IgA (2.16±0.11) g/L均升高(χ^2=4.427,P<0.05;t=15.218,P<0.05;t=10.125,P<0.05);SLE患者外周血B细胞比例[(6.78±0.29)%]、miRNA-21-5p表达水平(7.52±0.59)%较健康对照外周血B细胞比例[(2.03±0.24)%]、miRNA-21-5p表达水平(3.60±0.62)均升高(t=59.064,P<0.05;t=19.317,P<0.05),而SLE患者外周血PDCD4基因表达水平(1.54±0.35)较健康对照(4.42±0.42)降低(t=19.126,P<0.05);与空白对照组和miRNA-21-5p NC组相比,miRNA-21-5p Inhibitor组细胞增殖受到抑制,细胞凋亡比例升高(F=5.244,P<0.05;F=37.903,P<0.05);与空白对照组和PDCD4 NC组相比,PDCD4 siRNA组细胞增殖能力增强,细胞凋亡比例减少(F=5.956,P<0.05;F=25.431,P<0.05).双荧光素酶报告基因实验分析结果显示,PDCD4是miRNA-21-5p的靶基因.结论 miRNA-21-5p可能通过抑制PDCD4的表达促进SLE患者外周血B细胞增殖,导致B细胞凋亡异常.miRNA-21-5p可作为SLE治疗新的靶向基因.     

人胶质瘤细胞及组织中miRNA-221、miRNA-222表达变化及意义

目的观察人胶质瘤细胞及组织中miRNA-221、miRNA-222的表达变化,探讨其与肿瘤放射敏感性的相关性。方法根据Trizol试剂盒说明书,从体外培养的人胶质瘤细胞系U87、U251、A172、LN229和60例胶质瘤组织(手术切除获得)中提取其总RNA,采用RT-PCR法测定miRNA-221及miRNA-222,以正常脑组织为对照。60例胶质瘤患者术后均予以放疗,治疗后MRI检查随访,分析胶质瘤患者的放疗疗效。采用克隆形成实验测算X线照射后(照射剂量为2 Gy)的胶质瘤细胞存活率(SF),以胃黏膜上皮细胞(ges)为对照。结果与对照组织及细胞相比,胶质瘤组织及细胞中miRNA-221、miRNA-222呈高表达(P<0.05),其中高度恶性胶质瘤组织中miRNA-221、miRNA-222均明显高于低度恶性者(P<0.05)。X线照射后胶质瘤细胞SF明显高于ges(P均<0.05),胶质瘤各细胞中miRNA-221、miRNA-222表达量与X线照射后胶质瘤SF呈正相关(r=0.673、0.696,P均<0.01)。胶质瘤组织中miRNA-221、miRNA-222高表达的患者较低表达者生存时间短(P均<0.05)。结论人胶质瘤细胞及组织中miRNA-221、miRNA-222均呈高表达。miRNA-221、miRNA-222表达量与胶质瘤细胞的放射抗性呈正相关关系,胶质瘤组织中miRNA-221、miRNA-222表达量与患者生存时间呈负相关关系。     

年龄相关性白内障晶状体上皮细胞中miR-181a和SIRT1的表达关系及意义

目的探讨白内障晶状体上皮细胞微小RNA-181a(miR-181a)与沉默信息调节因子(SIRT)1的表达关系,研究其在年龄相关性白内障患者的诊治及病情评估方面的临床意义。方法选取诊断为年龄相关性白内障患者62例为白内障组,另选取同期近视矫正手术患者42例为对照组,利用RT q-PCR法,检测两组晶状体上皮细胞miR-181a与SIRT1表达水平;Pearson法测定miR-181a与SIRT1相关性;利用Lipofectamine 2000转染miR-181a模拟物和抑制剂,调节人晶状体上皮细胞miR-181a表达水平,RT q-PCR法检测SIRT1表达水平;酶联免疫吸附试验(ELISA)检测晶状体上皮细胞中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱氨酸(GSH)、谷胱氨酸过氧化物酶(GSH-Px)水平。结果与对照组相比,白内障组miR-181a表达水平显著升高,SIRT1表达水平显著降低(P<0.05);Pearson相关分析显示,miR-181a和SIRT1呈负相关关系(r=-0.719,P<0.05);miR-181a模拟物组SIRT1表达水平显著低于模拟物阳性对照组(P<0.05),miR-181a抑制剂组,SIRT1表达水平显著高于抑制剂阴性对照组(P<0.05);白内障组SOD、GSH、GSH-Px水平明显低于正常组(P<0.05),MDA水平明显高于正常组(P<0.05);miR-181a表达水平与SOD、GSH、GSH-Px因子呈负相关关系(r=-0.702,-0.739,-0.776),与MDA水平呈正相关关系(r=0.679);SIRT1表达水平与SOD、GSH、GSH-Px呈正相关关系(r=0.699,0.743,0.763),与MDA水平呈负相关关系(r=-0.684)。结论白内障晶状体上皮细胞中miR-181a高表达,SIRT1低表达,miR-181a可调控人晶状体上皮细胞中SIRT1表达;SIRT1低表达可促使晶状体上皮细胞凋亡,从而影响或导致了年龄相关性白内障的发病。