miR-122在肝癌细胞中的表达及其与细胞周期调控的关系

目的 探讨miR-122在肝癌细胞中的表达及其与细胞周期调控间的关系。方法 应用荧光定量PCR和原位杂交法分别检测50例肝癌及相应癌旁组织中miR-122的表达水平;用流式细胞术检测肝癌细胞的细胞周期;分析miR-122的表达水平与细胞周期的关系。结果 50例肝癌组织中miR-122的相对表达量为癌旁正常组织的（17.74±9.20）％。其中,40例高中分化肝癌组织中miR-122的相对表达量为癌旁组织的（19.89±8.80）％,而在10例低分化肝癌中仅为癌旁组织的（9.16±4.77）％。肝癌组织中miR-122的阳性表达率为30.0％（15／50）,癌旁组织为80.0％（40／50）,组间差异有统计学意义（P＜0.05）。与癌旁组织相比,肝癌细胞G1／G0期比例降低（P＜0.05）,S期比例增加;与miR-122阳性表达组比较,miR-122阴性表达组的肝癌细胞S期比例增加,而G1／G0期细胞比例降低（P＜0.05）。结论 miR-122在肝癌组织中低表达,且表达水平随着肿瘤分化程度的降低而下调,这种改变促进肝癌细胞的分裂和增殖。     

应用实时荧光定量PCR检测血清miR-122和miR-22及其在慢性乙型肝炎患者中的表达

目的 建立血清中microRNA(miRNA)的实时荧光定量PCR(qRT-PCR)的检测方法,对慢性乙型肝炎患者血清中miR-122和miR-22的表达水平进行检测和分析,探讨其在慢性乙型肝炎患者血清中表达的意义。 方法 茎环引物用于成熟型miRNA反转录,以建立SYBR Green Ⅰ PCR筛选和定量检测miRNA的方法。同时,采用10倍梯度稀释的miR-122标准品cDNA(1~10⁹拷贝/微升)评价其敏感度;采用熔解曲线评价其检测miRNA的特异性;通过对2×10⁵、2×10⁶、2×10⁷拷贝/微升的miR-122标准品cDNA分别进行批内20次重复实验,以其循环阈值(Ct)的变异系数(CV)评价其精密度。采用建立的qRT-PCR技术检测慢性乙型肝炎患者及正常人血清中miR-122和miR-22的表达水平。 结果 建立了茎环引物的RT-PCR法检测血清中的miRNA,该方法的线性范围宽,敏感度高,重复性好,方法简便。在慢性乙型肝炎患者组中血清miR-122和miR-22的相对表达量分别为17.88和5.35;与正常对照组中血清miR-122和miR-22的相对表达量(分别为1.80和1.67)比较,差异有统计学意义(P均=0.000)。 结论 建立了茎环引物RT-PCR实时荧光定量PCR方法检测血清中的miR-122和miR-22,血清miR-122和miR-22在慢性乙型肝炎患者的血清中表达显著升高。     

Prior nasal delivery of antagomiR-122 prevents radiation-induced brain injury

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Prevalence of humoral immunodeficiency in adult patients with recurrent tonsillitis

PurposeRecurrent tonsillitis in adults has a significant impact on patients' daily life and healthcare costs. Humoral immunodeficiency increases the susceptibility to recurrent infections. The purpose of this study was to investigate the prevalence and contribution of humoral immunodeficiency in adult patients with recurrent tonsillitis.Material and methodsA prospective cross-sectional study conducted over 3 years duration with two groups of subjects. Group 1: included 50 normal adult subjects and group 2: included 50 adult patients with recurrent tonsillitis. Recruitment occurred in a tertiary care hospital in Egypt. Different immunoglobulins (Ig A, Ig M and Ig G isotypes) were quantitatively assessed and compared in 2 groups. Incidence of different infections was also compared in patients with humoral immunodeficiency versus patients with intact immunity.Results4 (8%) subjects in group 1 had selective humoral Immunodeficiency versus 13 (26%) patients in group 2. Patients with recurrent tonsillitis had significantly lower mean of most assessed immunoglobulins: IgA (P = 0.002), IgM (P = 0.003), IgG (P < 0.0001), IgG1 (P < 0.0001) and IgG3 (P < 0.0001) compared to normal subjects; with no significant difference in mean of IgG2 (P = 0.395) and IgG4 (P = 0.105). Patients with humoral immunodeficiency had significantly higher incidence of tonsillitis (P < 0.0001) and rhinosinusitis (P < 0.0001) attacks compared to patients with normal immunity.ConclusionAdult patients with recurrent tonsillitis may have higher prevalence of humoral immunodeficiency compared to normal subjects. These findings suggest that assessment of immune function should be undertaken routinely in these patients.     

miR-122通过靶向RUNX2诱导胶质瘤细胞凋亡的实验研究

目的 探讨微小RNA122（miR-122）对神经胶质瘤细胞凋亡的影响及可能分子机制。方法 采用荧光定量PCR（QPCR）检测人正常星形胶质细胞系HA1800和胶质瘤细胞系U251、U87、SHG44、MO59K中miR-122的表达情况。双荧光素酶报告实验评价miR-122对RUNX2的靶向调控作用。向U251细胞转染miR-122 mimics（miR-122组）和阴性对照NC（NC组），流式细胞术检测细胞凋亡，Western blotting检测凋亡相关蛋白Bcl-2、Bax、cleaved caspase-3的表达。向U251细胞同时转染miR-122 mimics和siRUNX2（miR-122+siRUNX2组），流式细胞术检测细胞凋亡。结果 U251、U87、SHG44、MO59K细胞系中miR-122表达水平分别为0.34±0.052、0.65±0.061、0.59±0.071和0.69±0.098，显著低于正常星形胶质细胞系HA1800的1.17±0.173，差异具有统计学意义（P＜0.05）。miR-122可抑制野生型RUNX2 3’UTR报告基因载体的荧光素酶活性，而对突变型RUNX2 3’UTR-MUT的荧光素酶活性无影响。miR-122组U251细胞凋亡率为（23.77±0.83）％，高于NC组的（6.17±0.12）％，差异具有统计学意义（P＜0.05）；低于miR-122+siRUNX2组的（70.85±0.35）％（P＜0.05）。Western blotting检测结果显示，miR-122组RUNX2蛋白表达量为0.57±0.048，低于NC组的1.21±0.19（P＜0.05）。与NC组比较，miR-122组Bax（3.47±0.077）、 cleaved caspase-3（4.38±0.121）表达均明显上调，而Bcl-2（0.39±0.104）明显下调（P＜0.05）。结论 miR-122可以通过靶向RUNX2促进线粒体凋亡通路的激活，促进神经胶质瘤细胞凋亡。     

血清miR-122和miR-570联合检测对原发性肝癌的临床诊断价值

目的:检测原发性肝癌(PHC)患者血清中miR-122和miR-570的表达水平及临床应用价值。方法:选择2017年2月至2019年1月我院收治的30例原发性肝癌患者作为研究组,选择我院同期30例健康体检者为对照组。采用实时荧光定量PCR方法检测血清miR-122和miR-570的表达水平,利用受试者工作特征曲线评估血清miR-122和miR-570的诊断效能。结果:miR-122和miR-570在原发性肝癌患者血清的表达水平显著低于健康对照者的表达水平,差异具有统计学意义（均P<0.01）;miR-122表达水平与原发性肝癌TNM分期、病理分级、原发性肝癌远处转移程度呈负相关（P<0.05）,miR-570表达水平与原发性肝癌TNM分期、病理分级、原发性肝癌远处转移程度无相关性;miR-122诊断原发性肝癌的AUC为0.816,敏感度为80.5%,特异度为 90.1%;miR-570诊断原发性肝癌的AUC为0.862,敏感度为83.3%,特异性为90.0%;联合血清miR-122和miR-570诊断原发性肝癌的AUC为0.903,敏感性为90.0%,特异性为83.6%。结论:原发性肝癌患者血清miR-122和miR-570降低,血清miR-122表达水平与原发性肝癌TNM分期、病理分级、原发性肝癌远处转移程度呈负相关,联合检测血清miR-122和miR-570对于原发性肝癌的检出具有较高的参考价值。