miR-375在变应性鼻炎小鼠鼻黏膜上皮细胞凋亡中的调控作用

目的探讨miR-375在变应性鼻炎小鼠鼻黏膜上皮细胞凋亡和炎症反应中的调控作用。方法运用卵清蛋白(OVA)致敏的小鼠变应性鼻炎模型,使用实时定量PCR(qRT-PCR)、蛋白质印迹试验(Western Blot)、酶联免疫吸附试验(ELISA)、免疫组织化学检测鼻黏膜上皮细胞内miR-375、JAK2、细胞凋亡相关蛋白(JAK2蛋白,裂解的蛋白酶3(Cleaved caspase 3),聚[ADP-核糖]聚合酶裂解酶(Cleaved PARP),蛋白酶3(Caspase 3),聚[ADP-核糖]聚合酶(PARP),p-STAT3蛋白,STAT3蛋白和β肌动蛋白(β-actin))和血浆IL-6、TNF-α、IL-10的表达水平。结果miR-375在变应性鼻炎小鼠的鼻黏膜上皮细胞中表达降低,而JAK2表达增高;JAK2蛋白、p-STAT3蛋白和裂解的蛋白酶3均在OVA组表达增高;给OVA致敏的变应性鼻炎小鼠注射miR-375模拟物可以导致血清IL-6、TNF-α的分泌下降,而IL-10分泌增加,该作用可以被带有过表达JAK2的腺病毒感染后而减弱。结论miR-375/JAK2调控通路存在于变应性鼻炎鼻黏膜上皮细胞中,并通过JAK2/STAT3信号通路调控细胞的凋亡和炎症反应,miR-375在变应性鼻炎的病程中有保护性机制。     

miR-31靶向抑制IL1RN促进HaCaT细胞中IL-1β的分泌

目的研究miR-31/IL1RN途径在银屑病发生发展中的功能作用。方法分析miR-31在银屑病皮损组织及其对照正常组织中的表达差异,然后在HaCaT细胞中研究miR-31表达变化对IL-1Ra蛋白表达的影响,双荧光素酶实验验证miR-31与IL1RN mRNA 3′UTR的靶向结合关系,最后检测miR-31和IL1RN基因对炎症因子IL-1β分泌的影响。结果 miR-31在银屑病皮损组织中的表达高于非皮损组织,而体外细胞实验显示,miR-31过表达可抑制细胞中IL-1Ra蛋白表达,干扰miR-31则可上调细胞中IL-1Ra蛋白表达。双荧光素酶实验结果表明,miR-31可靶向结合IL1RN的3′UTR区。功能实验结果显示,miR-31过表达可促进IL-1β的分泌,而IL1RN过表达则可下调IL-1β的分泌,并可削弱miR-31对IL-1β分泌的促进作用。结论 miR-31通过靶向IL1RN的3′UTR区域抑制IL-1Ra的表达,促进IL-1β分泌,从而参与银屑病的进展。     

肿瘤相关巨噬细胞相关性miR-99a对子宫内膜癌细胞生长和侵袭的调控作用

背景与目的：肿瘤相关巨噬细胞（tumor-associated macrophage，TAM）浸润与肿瘤进展密切相关，但作用机制尚不明确，因此，探索miR-99a对单核巨噬细胞极化的影响及其对子宫内膜癌（endometrial cancer，EC）细胞生长、侵袭的影响。方法：检测EC组织中巨噬唾液酸蛋白（macrosialin）CD68表达并分析其与临床病理学特征之间的关系；运用人EC细胞系HEC-1B、RL95-2培养上清液诱导人单核细胞U937向TAM（M2型巨噬细胞）分化；将人工合成的miR-99a模拟物片段转染至诱导后的巨噬细胞，转染后运用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）及流式细胞术检测巨噬细胞相关因子CD68、CD163以及巨噬细胞甘露糖受体（mcrophage mannose receptor）CD206表达量变化，并运用酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测巨噬细胞分泌相关细胞因子IL-12、IL-4和IL-10分泌量变化；将转染miR-99a的诱导后巨噬细胞与EC细胞共培养，运用细胞计数试剂盒（cell counting kit-8，CCK-8）和transwell法检测其对EC细胞增殖和侵袭能力的影响，并初步分析其可能的作用机制。结果：EC组织CD68高表达并与肿瘤肌层浸润及血管生成呈正相关；肿瘤细胞培养上清液成功诱导单核细胞向M2型TAM极化。转染miR-99a后单核细胞组CD68及CD163表达较对照组下降（P＜0.01），而CD206表达差异无统计学意义（P＞0.05），流式细胞术进一步证实上述表达变化；ELISA结果发现，转染miR-99a诱导后巨噬细胞中IL-12分泌增多（P＜0.01），而IL-4、IL-10分泌减少（P＜0.01），提示巨噬细胞向M2型极化受抑制。将诱导后巨噬细胞与EC细胞共培养，共培养后EC细胞增殖侵袭能力较对照组增加，而转染miR-99a模拟片段至诱导后巨噬细胞能够抑制其对增殖（P＜0.01）及侵袭能力的促进作用（P＜0.05）。诱导后巨噬细胞中过表达miR-99a后细胞中mTOR及其通路受到抑制。结论：EC间质巨噬细胞浸润与肿瘤肌层浸润及血管新生相关，miR-99a能够逆转单核细胞向M2表型极化，并抑制EC细胞介导TAM的促EC细胞生长和侵袭作用，其作用可能通过调控mTOR通路产生。     

Allogeneic stem cell transplantation in AML with t(6;9)(p23;q34);DEK-NUP214 shows a favourable outcome when performed in first complete remission

Acute myeloid leukaemia (AML) with t(6;9)(p23;q34) is a poor-risk entity, commonly associated with FLT3-ITD (internal tandem duplication). Allogeneic stem-cell tranplantation (allo-SCT) is recommended, although studies analysing the outcome of allo-SCT in this setting are lacking. We selected 195 patients with t(6;9) AML, who received a first allo-SCT between 2000 and 2016 from the EBMT (European Society for Blood and Marrow Transplantation) registry. Disease status at time of allo-SCT was the strongest independent prognostic factor, with a two-year leukaemia-free survival and relapse incidence of 57% and 19% in patients in CR1 (first complete remission), 34% and 33% in CR2 (second complete remission), and 24% and 49% in patients not in remission, respectively (P < 0·001). This study, which represents the largest one available in t(6;9) AML, supports the recommendation to submit these patients to allo-SCT in CR1.     

颅内动脉瘤破裂出血术后血清HIF-1α、miR-210表达与脑血管痉挛的关系

目的检测颅内动脉瘤破裂出血术后血清缺氧诱导因子1α（HIF-1α）、miR-210表达情况，并探讨其与脑血管痉挛（CVS）的关系。 方法选取禹城市人民医院神经外科自2015年1月至2018年12月收治的87例颅内动脉瘤破裂急诊自发性蛛网膜下腔出血（SAH）并接受介入栓塞或开颅夹闭治疗的患者为研究对象，采用头颅X线、CT及全脑数字减影血管造影（DSA）检查判断CVS的发生情况，并评估其严重程度。术后3、7 d采用ELISA法检测血清HIF-1α表达情况，采用qRT-PCR法检测血清miR-210表达情况。分析颅内动脉瘤破裂出血患者术后血清HIF-1α、miR-210表达的关系，采用ROC曲线分析颅内动脉瘤破裂出血患者术后3 d血清HIF-1α、miR-210水平对CVS的诊断价值。 结果87例颅内动脉瘤破裂出血患者出现术后CVS者37例（42.53%），其中轻度CVS 10例（11.49%），中度19例（21.84%），重度8例（90.20%）；术后3、7 d，与无CVS组患者相比，不同程度CVS患者血清中的HIF-1α、miR-210水平均显著升高（P<0.05），且CVS程度越重，血清HIF-1α、miR-210表达水平越高，不同程度CVS患者术后3、7 d时血清HIF-1α、miR-210水平均显著高于术前（P<0.05），术后7 d时血清HIF-1α、miR-210水平均显著低于术后3 d，差异有统计学意义（P<0.05）；颅内动脉瘤破裂出血术后3、7 d患者血清HIF-1α水平与miR-210水平均呈正相关（r=0.381、0.631，P<0.05）；术后3 d血清HIF-1α、miR-210水平及HIF-1α+miR-210联合诊断颅内动脉瘤破裂出血患者CVS的曲线下面积分别为0.834、0.769、0.900，二者联合诊断CVS的敏感度为93.06%，准确度为87.36%，均高于单项指标检测。 结论颅内动脉瘤破裂出血术后血清HIF-1α、miR-210水平可有效预示CVS的发生，二者可能成为CVS发生、发展的重要生物学指标。     

CircKIF4A靶向调控miR-384表达对卵巢癌SKOV3细胞增殖和凋亡的影响

Objective?To explore the effect of circKIF4A on the proliferation and apoptosis of ovarian cancer SKOV3 cells and its regulatory mechanism on miR-384. Methods?The qRT-PCR method was used to detect the expression of circKIF4A and miR-384 in ovarian cancer tissues and adjacent tissues. si-NC, si-circKIF4A, miR-NC, miR-384 mimics, si-circKIF4A and anti-miR-NC, si-circKIF4A and anti-miR-384 were transfected into SKOV3 cells respectively. The qRT-PCR method was used to detect the expression of circKIF4A and miR-384 in SKOV3 cells. MTT experiment and flow cytometry experiment were used to detect cell proliferation and apoptosis respectively. The dual luciferase reporter experiment was used to detect the targeting relationship between circKIF4A and miR-384. Results?Compared with adjacent tissues, the expression level of circKIF4A in ovarian cancer tissues was increased [(1.00±0.06), (4.28±0.32)] (t=62.915, P＜0.05), and the expression level of miR-384 was decreased [(1.00± 0.05), (0.43±0.03)] (t=61.047, P<0.05). After transfection with si-circKIF4A, the OD value of SKOV3 cells was decreased [(0.75±0.05), (0.41±0.03)] (t=17.493, P＜0.05), and the apoptosis rate was increased [(6.36±0.53)%, (23.19± 2.21)%] (t=22.216, P<0.05). After transfection of miR-384 mimics, the OD value of SKOV3 cells was decreased [(0.73±0.05), (0.47±0.04)] (t=12.182, P＜0.05), and the apoptosis rate was increased [(7.53±0.41)%, (19.11)±1.06)%] (t=30.567, P<0.05). The dual luciferase report experiment confirmed that circKIF4A could adsorb miR-384 and can act as a sponge molecule for miR-384. After co-transfection with si-circKIF4A and anti-miR-384, the OD value of SKOV3 cells was increased [(0.40±0.04), (0.65±0.03)] (t=15.000, P＜0.05), and the apoptosis rate was decreased [(25.20± 2.21)%, (10.37±0.86)%] (t=18.761, P＜0.05). Conclusion?Inhibition of circKIF4A expression could negatively regulate the expression of miR-384, thereby inhibiting the proliferation of ovarian cancer cells and inducing apoptosis.     

lncRNA CYTOR靶向调控miR-503/CCNE1信号轴促进上皮性卵巢癌细胞增殖北大核心

目的:探讨细胞骨架调节剂(lncRNA cytoskeleton regulator,CYTOR)是否通过靶向调控miR-503来促进细胞周期蛋白E1(cyclin E1,CCNE1)影响上皮性卵巢癌增殖。方法:实时定量聚合酶链反应(RT-qPCR)检测CYTOR在不同临床分期、病理分级和有无淋巴结转移患者人上皮性卵巢癌组织、癌旁组织和正常组织以及不同卵巢癌细胞中的表达;starBase数据库(网址:http://starbase.sysu.edu.cn/)预测分析CYTOR和miR-503以及miR-503与CCNE1之间的关系,同时荧光素酶实验进行验证;RT-qPCR和免疫荧光分别检测miR-503与CCNE1在人上皮性卵巢癌组织、癌旁组织和正常组织以及不同卵巢癌细胞中的表达;RNA转染技术在A2780中细胞分别沉默CYTOR、过表达miR-503和沉默CCNE1表达,CCK-8检测各组细胞增殖能力变化;RT-qPCR和免疫荧光检测沉默CYTOR后miR-503和CCNE1的表达以及过表达miR-503后CCNE1的表达。结果:CYTOR和CCNE1在上皮性卵巢癌组织中表达增加,同时在临床分期III-IV期、病理分级G_(3)级、淋巴结转移阳性患者肿瘤组织中表达同样显著增加且导致卵巢癌患者术后预后较差;通过数据库分析CYTOR可直接靶向抑制miR-503,而miR-503可直接靶向抑制CCNE1表达,双荧光素酶实验结果与之相同;CYTOR和CCNE1促进上皮性卵巢癌增殖,而miR-503抑制上皮性卵巢癌增殖作用。结论:lncRNA CYTOR通过调控miR-503/CCNE1轴促进上皮性卵巢癌的增殖。     

miR-372 inhibits p62 in head and neck squamous cell carcinoma in vitro and in vivo

Here we showed that exogenous miR-372 expression and knockdown of p62 (sequestosome1 or SQSTM1), both increased migration of head and neck squamous cell carcinoma (HNSCC) cells. p62 induced phase II detoxification enzyme NADPH quinone oxidoreductase 1 (NQO1), which decreased ROS levels and cell migration. Also, miR-372 decreased p62 during hypoxia, thus increasing cell migration. Levels of miR-372 and p62 inversely correlated in human HNSCC tissues. Plasma levels of miR-372 was associated with advanced tumor stage and patient mortality. Both plasma and salivary miR-372 levels were decreased after tumor resection. We conclude that miR-372 decreases p62, thus increasing ROS and motility in HNSCC cells.     

微小RNA在人类压疮组织中的异常表达

目的建立人类压疮组织微小RNA表达谱。方法收集本院24例压疮组织临床样本,选取其中4例用于压疮微小RNA芯片制备。采用生物信息学算法比较压疮组织与正常组织中差异表达的微小RNA,获得压疮微小RNA表达谱。进一步利用20例测试样本,采用实时荧光定量-逆转录-聚合酶链反应对所鉴定的表达谱进行验证。结果获得了12个差异表达的微小RNA,其中包括5个表达水平上调和7个表达水平下调的微小RNA,其中表达水平下调的miR-17差异表达水平最显著。结论作者建立了一组由12个微小RNA组成的压疮组织微小RNA表达谱,为压疮发生及发展分子调控机制研究提供了丰富的素材。