系统性红斑狼疮患者CD4+ T细胞中miR-17和miR-20a 的表达及与病情活动的相关性研究

【摘要】 目的 探讨miR-17和miR-20a在系统性红斑狼疮（SLE）患者CD4+ T细胞中的表达及与疾病活动的相关性。 方法 应用实时荧光定量聚合酶链反应（PCR）检测SLE患者活动组17例、缓解组13例及健康对照组18例外周血CD4+ T细胞中miR-17和miR-20a的表达水平,并与SLE患者临床指标进行相关性分析。 结果 SLE患者组30例miR-17的表达水平（0.24 ± 0.08）显著高于健康对照组（0.15 ± 0.06）,两组比较,P < 0.05;两组miR-20a的表达水平分别为0.19 ± 0.10和0.12 ± 0.09,两组比较,P < 0.05。SLE活动组、缓解组与健康对照组miR-17表达水平分别为0.27 ± 0.07、0.20 ± 0.05、0.16 ± 0.08,活动组高于缓解组（P < 0.05）和健康对照组（P < 0.05）;miR-20a表达水平三组分别为0.22 ± 0.10、0.15 ± 0.07、0.13 ± 0.09,活动组高于缓解组（P < 0.05）和健康对照组（P < 0.05）;缓解组miR-17和miR-20a的表达水平与健康对照组比较差异均无统计学意义（P > 0.05）。SLE患者组miR-17、miR-20a的表达与SLE活动指数（SLEDAI）均呈正相关（r = 0.45,P < 0.05和r = 0.38,P < 0.05）,与抗dsDNA抗体滴度呈正相关（r = 0.54,P < 0.01和r = 0.46,P < 0.01）,而与C3水平呈负相关（r = -0.43,P < 0.05和r = -0.42,P < 0.05）。 结论 SLE患者外周血CD4+ T细胞中miR-17、miR-20a表达上调,且miR-17、miR-20a表达水平与SLE病情活动相关。     

miRNA-1297 Induces Cell Proliferation by Targeting Phosphatase and Tensin Homolog in Testicular Germ Cell Tumor Cells

To investigate the role of miR-1297 and the tumor suppressor gene PTEN in cell proliferation of testicular germ cell tumors (TGCT). MTT assays were used to test the effect of miR-1297 on proliferation of the NCCIT testicular germ cell tumor cell line. In NCCIT cells, the expression of PTEN was assessed by Western blotting further. In order to confirm target association between miR-1297 and 3’-UTR of PTEN, a luciferase reporter activity assay was employed. Moreover, roles of PTEN in proliferation of NCCIT cells were evaluated by transfection of PTEN siRNA. Proliferation of NCCIT cells was promoted by miR-1297 in a concentration-dependent manner. In addition, miR-1297 could bind to the 3’-UTR of PTEN based on luciferase reporter activity assay, and reduced expression of PTEN at protein level was found. Proliferation of NCCIT cells was significantly enhanced afterknockdown of PTEN by siRNA. miR-1297 as a potential oncogene could induce cell proliferation by targeting PTEN in NCCIT cells.     

细胞外信号调节激酶/miR-133b在甲基苯丙胺致PC12细胞神经毒性的作用及机制

目的 探讨甲基苯丙胺(MA)致神经细胞毒性过程中细胞外信号调节激酶(ERK)和miR-133b的表达变化及调控机制。方法 用MA建立PC12神经细胞损伤模型,采用四甲基偶氮唑盐(MTT)检测细胞活性及镜下形态观察确定MA最佳损伤浓度;应用流式细胞术检测细胞内活性氧（ROS）水平;通过 Western blotting技术测定总ERK1/2和磷酸化ERK1/2 (p-ERK1/2)的表达变化;并应用实时定量聚合酶链反应（Real-time PCR）测定miR-133b的表达变化。为进一步分析ERK/miR133b分子通路的作用关系,经U0126特异阻断ERK通路,检测miR-133b的表达变化。 结果 给予不同浓度的MA,均可导致PC12细胞损伤,其中800μmol/L MA处理后,大部分胞体变圆,神经突起退缩,神经网络消失。MTT结果显示细胞活性明显下降。进一步的细胞毒性机制分析显示,MA处理后,细胞内ROS水平升高,p-ERK表达增高,miR-133b表达降低;并且给予ERK通路抑制剂U0126（10μmol/L）后,miR-133b表达升高,细胞活性增强,胞内ROS水平降低,镜下细胞损伤改善。 结论 MA可通过上调ERK磷酸化抑制miR-133b表达,介导神经元毒性损伤。     

Expression of MiR200a,miR93, Metastasis-related GeneRECK and MMP2/MMP9 in Human Cervical Carcinoma -Relationship with Prognosis

Aim and Background: Cervical cancer remains the third most common cancer in women globally afterbreast and colorectal cancer. Well-characterized biomarkers are necessary for early diagnosis and to predictmetastatic progression and effective therapy. MiRNAs can regulate gene expression, cell growth, differentiationand apoptosis by targeting mRNAs for translational repression or degradation in tumor cells. The present studywas conducted to assess expression of miR93, miR200a, RECK, MMP2, MMP9 in invasive cervical carcinoma,and analyze their clinical significance. Method: A total of 116 patients with invasive cervical carcinoma and 100patients undergoing hysterectomy for benign lesions were retrospectively examined. Quantitative real-time PCRwas performed to determine expression of miR93 and miR200a while RECK, MMP2, MMP9 and MVD wereassessed by immunohistochemical staining. Results: Cervical carcinoma patients demonstrated up-regulationof miR-93, miR-200a, MMP2 and MMP9, with down-regulation of RECK as compared to benign lesion tissues.RECK was significantly inversely related to invasion and lymphatic metastasis. The 5-year survival rate forpatients with strong RECK expression was significantly higher than that with weakly expressing tumors.Conclusion: MiR-93 and miR-200a are associated with metastasis and invasion of cervical carcinoma. Thustogether with RECK they are potential prognostic markers for cervical carcinoma. RECK cooperating withMMP2, MMP9 expression is a significant prognostic factor correlated with long-term survival for patients withinvasive cervical carcinoma.     

Hsa-miR-181a-5p Expression and Effects on Cell Proliferation in Gastric Cancer

Purpose: MicroRNAs (miRNAs) are small endogenous, non-coding, single-stranded RNAs (approximately22 nt). Accumulating evidence has shown that aberrant miRNA expression is pronounced and correlatedwith gastric cancer genesis and progression. Materials and Methods: Expression levels of miR-181a-5p in GCtissues and cell lines were assessed by qRT-PCR and tested for correlation with clinical features. In addition,effects of miR-181a-5p on GC cell growth were investigated. Results: Our findings indicate that miR-181a-5pis upregulated in GC, in correlation with lymph node invasion, nerve invasion and vascular invasion (P<0.05).Enforced expression of miR-181a -5p promoted cell proliferation ability. Conclusions: This study suggested thatincreased miR-181a-5p is related to GC progression. MiR-181a-5p may represent a potential therapeutic targetfor GC.     

PRL-3调节PI3K信号通路在促进结肠癌细胞增殖侵袭中的作用

【目的】 研究肝再生磷酸酶-3 (PRL-3)的过表达与PI3K信号通路的调节及其与结肠癌细胞增殖侵袭的关系?【方法】 构建稳定转染PRL-3基因和空白对照质粒的结肠癌细胞株LoVo- PRL-3和LoVo-VC,用qRT-PCR和Western blot的方法对miR-21及其靶蛋白PTEN的表达进行检测,在稳转细胞株中转染miR-21或对其进行敲除,用CCK-8?Tanswell实验对细胞的增殖侵袭能力的变化进行研究?【结果】 LoVo- PRL-3细胞的增殖侵袭能力要强于对照组LoVo-VC细胞?在结肠癌细胞株LoVo-PRL-3中miR-21的表达上调而PTEN的表达被下调,在LoVo-VC细胞中过表达miR-21促进了细胞的增殖侵袭并降低PTEN的表达,而在LoVo- PRL-3细胞中敲除miR-21抑制了细胞的增殖侵袭并能部分恢复PTEN的表达?【结论】 PRL-3通过上调miR-21抑制PTEN的表达,调节了PI3K信号通路,从而促进了结肠癌细胞的增殖侵袭?     

特应性皮炎患者外周血调节性T细胞及皮损中miR-31、Foxp3的表达

目的 探讨特应性皮炎（AD） 患者外周血调节性T细胞（Treg 细胞）及皮损组织中miR-31、Foxp3的表达水平及其相关性。方法 用流式细胞仪检测37例AD患者外周血Treg细胞的比例,实时荧光定量RT-PCR检测miR-31及Treg细胞特异性转录因子Foxp3 mRNA的表达水平;并以33例性别、年龄匹配的健康儿童做对照。检测5例急性期AD患者皮损与皮损周围组织中miR-31及Foxp3 mRNA的表达水平,并以5例健康人皮肤组织标本为对照。采用独立样本t检验、单因素方差分析和线性相关进行统计学分析。结果 AD患者外周血Treg细胞比例明显低于健康对照组（2.12% ± 0.60%比4.99% ± 1.27%,P < 0.01）;miR-31 mRNA表达水平明显高于健康对照组（6.01 ± 1.76比1.62 ± 0.51,P < 0.01）;Foxp3 mRNA表达水平显著低于健康对照组（0.78 ± 0.17比1.87 ± 0.71,P < 0.01）。AD患者皮损、皮损周围组织及健康人皮肤组织中miR-31及Foxp3 mRNA表达水平间差异均有统计学意义。miR-31表达水平：皮损 > 皮损周围组织 > 健康人皮肤组织,F = 54.501,P < 0.01;Foxp3表达水平：皮损 < 皮损周围组织 < 健康人皮肤组织,F = 37.837,P < 0.01。AD患者外周血miR-31 mRNA表达水平与疾病严重程度评分呈正相关（r = 0.417,P = 0.010）,与Treg细胞比例（r = -0.404,P = 0.013）及Foxp3 mRNA表达水平呈负相关（r = -0.409,P = 0.012）;皮损及皮损周围组织中miR-31 mRNA表达水平与Foxp3 mRNA表达水平呈负相关（r = -0.392,P = 0.032）。结论 AD患者miR-31高表达及Foxp3低表达可能与AD的发病相关。 【关键词】 皮炎,特应性; miR-31; T淋巴细胞,调节性; Foxp3     

miR-16对大肠癌细胞增殖能力的影响

目的研究miR-16在大肠癌组织中的表达及其在大肠癌细胞株HCT116和LoVo细胞增殖中的作用。方法运用Realtime PCR法检测miR-16在33对配对肠癌及癌旁正常组织中的表达。HCT116和LoVo细胞转染miR-16 抑制剂、阴性对照剂和模拟剂;CCK8法检测细胞增殖;流式细胞仪进行细胞周期检测;Western blot法检测细胞中cyclinD1、CDK6和GAPDH蛋白表达。结果（1）Realtime PCR结果显示miR-16在大肠癌组织中低表达（P=0.047）。（2）miR-16过表达可抑制HCT116和LoVo细胞增殖,诱导细胞G₀/G₁期阻滞,降低cyclinD1和CDK6蛋白表达。结论miR-16过表达可以抑制大肠癌细胞的增殖能力。     

胃癌组织和循环miR-21与胃癌伊立替康敏感性关系的初步研究

背景与目的：肿瘤组织和细胞中的miR-21先后被证实与肿瘤对化疗药物的敏感性有关,高表达miR-21提示肿瘤对多种化疗药物不敏感。外周血蕴含丰富的肿瘤信息,为个体化药物治疗提供了新的检测来源。本研究初步探讨胃癌组织和血浆循环miR-21表达水平与胃癌铂类药物和伊立替康敏感性之间的关系。方法：系统收集35例经病理确诊的新鲜人胃癌标本,采用三维微组织块培养法行两种药物的体外敏感试验;实时荧光定量PCR检测血浆和对应胃癌石蜡组织中miR-21表达水平。结果：血浆miR-21与对应肿瘤组织miR-21呈正相关(rho=0.736, P＜0.001)。血浆miR-21水平与性别、年龄、病理类型、分化程度、淋巴结转移、TNM分期、肿瘤部位无统计学相关性,但是Ⅱ期和Ⅲ期的miR-21水平较Ⅰ期有偏高的趋势。铂类药物敏感组与耐药组肿瘤组织miR-21相对表达量分别为1.32(95%CI：0.73～1.90)和4.06(95%CI：1.71～6.41),差异无统计学意义(P=0.004);铂类药物敏感组与耐药组血浆miR-21相对表达量分别为5.25(95%CI：0.14～10.64)和5.82(95%,差异无统计学意义CI：2.27～9.37),差异无统计学意义(P=0.19)。伊立替康敏感组与耐药组肿瘤组织miR-21相对表达量分别为1.09(95%CI：0.65～1.54)和4.94(95%CI：2.44～7.44),差异无统计学意义(P<0.001);伊立替康敏感组与耐药组血浆miR-21相对表达量分别为1.86(95%CI：1.08～2.64)和12.42(95%CI：3.14～21.70)差异无统计学意义(P=0.001)。结论：血浆循环miR-21与对应肿瘤组织miR-21水平呈显著正相关,在一定程度上能反映肿瘤组织miR-21的水平。血浆miR-21水平与新鲜胃癌组织对伊立替康的敏感性呈负相关,肿瘤组织miR-21水平与新鲜胃癌组织对伊立替康和铂类药物的敏感性呈负相关。