反义miR-21通过RECK抑制胶质瘤细胞侵袭性生长的体内外研究

目的 探讨胶质瘤细胞LN229中RECK作为miR-21的调控靶点,在胶质瘤侵袭性生长中的作用.方法 将反义miR-21(AS-miR-21)寡核苷酸转染至人脑胶质瘤细胞LN229中.Real-time PCR检测LN229细胞中miR-21的表达量.荧光素酶实验检测miR-21对RECK的调控关系.Transwell实验评价LN229细胞侵袭能力的变化,应用Western blot检测细胞内MMP2/9和RECK蛋白水平的变化,ELISA实验检测培养基中活性MMP2/9的表达量,动物实验评价体内条件下肿瘤侵袭性的变化.结果 Real-time PCR显示转染组中miR-21的表达量与对照组相比下调60%.荧光素酶实验证明RECK是miR-21的靶点.Transwell实验证实胶质瘤细胞侵袭能力下降,Western blot和ELISA实验证实MMP2/9表达降低,动物实验及免疫荧光反映肿瘤侵袭性生长受抑制.结论 反义miR-21通过上调RECK的表达而抑制恶性胶质瘤细胞的侵袭性生长.

Abstract：

Objective To investigate the regulation of miR - 21 on invasion growth of human glioma cells by RECK.Method The human glioma LN229 cells were transfected with AS - miR - 21 or scrambled sequences by Lipofectamine2000.Real time PCR was conducted to detect the expression of miR-21.Luciferase experiment was performed to detect the relationship between miR-21 and RECK.The expression of RECK was evaluated by Western blot.The invasion ability was evaluated by transwell assay and subcutaneous models.Western Blot, ELISA and immunofluorescence were used to estimate the changes of MMP2/9.Results The expression of miR - 21 in LN229 cells decreased after transfection with AS-miR-21. It was proved that RECK was a direct target of miR -21 by luciferase experiment.Meanwhile, the high expression of RECK protein in AS - miR -21 group conformed its important function in this mechanism.Transwell assay demonstrated decreased invasion capability of LN229 cell lines transfected with AS- miR- 21.Western blot, ELISA, and immunofluorescence demonstrated the levels of MMP2/9 were down -regulated in AS -miR -21 group compared with control and scrambled group.Conclusions AS - miR -21 could depress the invasion of glioma cells owing to up - regulating the level of RECK which could inhibit MMP2/9 activities both in vitro and vivo.     

不同AKT1与AKT2表达状态的胶质瘤的生存分析

目的 探讨AKT1与AKT2在脑胶质瘤中的表达及与患者生存期的相关性.方法 免疫荧光法检测手术切除的50例胶质瘤组织标本中AKT1及AKT2的表达水平,确定两者表达的相关性并与生存期进行相关性分析.结果 Log - rank单因素检验,年龄≥50岁、KPS评分＜ 80分和WHO Ⅲ～Ⅳ级患者的生存期显著缩短.胶质瘤组织中AKT1及AKT2在WHOⅢ～Ⅳ级的表达率显著高于WHO Ⅰ～Ⅱ级,两者表达存在正相关,且与患者的生存期呈负相关.结论 AKT1与AKT2的表达状态对预测患者的生存期具有一定的临床价值.     

上海地区慢性荨麻疹患者变态反应原种类的临床检测分析

目的:研究上海地区慢性荨麻疹发病中常见变态反应原种类及其在不同年龄、性别患者中分布特点,为其预防和治疗提供依据。方法:应用阿罗格变态反应原点刺液对上海地区800例慢性荨麻疹患者进行皮肤点刺试验,按年龄将800例被调查者分为≤17岁、18～39岁、40～59岁和≥60岁组,分析30种常见变态反应原的阳性率,并比较其在不同性别及年龄患者中的阳性率。结果:皮肤点刺试验的总阳性率为49.38%,其中以粉尘螨和屋尘螨的阳性率最高,分别为33.38%和37.88%。仅对1种变态反应原有阳性反应者为1.00%,同时对2种或2种以上变态反应原有阳性反应者分别占10.25%及38.13%;男女间阳性反应率差异无统计学意义(P>0.05);随着患者年龄增大,粉尘螨和屋尘螨的阳性反应率呈明显下降趋势(P<0.01)。结论:上海地区慢性荨麻疹患者的变态反应原以粉尘螨和屋尘螨为主;部分变态反应原分布具有年龄特异性,提示不同年龄的患者在日常生活中应尽量避免接触相应变态反应原,以降低慢性荨麻疹发病率及复发率。