miR-221与miR-222通过靶基因TIMP3调控胶质瘤细胞侵袭能力

目的 探讨miR-221和miR-222在胶质瘤细胞侵袭过程中的作用及其机制.方法 采用反义寡核苷酸下调miR-221和miR-222表达,Transwell、Western blot、荧光素酶实验及体内实验分别检测细胞侵袭能力、体内肿瘤生长、相关基因蛋白表达变化及靶基因鉴定.结果 下调miR-221和miR-222表达能明显抑制胶质瘤细胞侵袭能力,同时相关侵袭蛋白MMP2和MMP9表达下降.miRNA靶基因预测软件分析、Western blot、荧光素酶实验证实TIMP3是miR-221和miR-222的靶基因.裸鼠皮下肿瘤模型显示下调miR-221和miR-222表达抑制体内肿瘤生长.免疫组化发现TIMP3表达上调,MMP2和MMP9表达下调.结论 在胶质瘤细胞中,侵袭相关蛋白TIMP3是miR-221和miR-222的一个新靶基因.

Abstract：

Objective To investigate the role and mechanism of miR -221 and miR -222 in glioma cell invasion. Method After reduction of miR -221 and miR -222 by antisense oligonucleotides, cell invasion,invasion related - gene expression and target identification were determined by Transwell assay, Western blot analysis and luciferase reporter assay,respectively. Moreover,the effects of miR -221 and miR -222 on xenograft tumors in nude mice were also observed. Results Down - regulation of miR - 221 and miR - 222 decreased glioma cell invasion in vitro, and inhibited glioma growth in a subcutaneous mouse model. Furthermore,down -regulation of miR -221 and miR - 222 resulted in obvious inactivation of MMP2 and MMP9. Western blot analysis and luciferase reporter assay showed that the reduction of miR - 221 and miR -222 repressed TIMP3 protein expression and TIMP3 mRNA 3'UTR existed the biding sites of miR -221 and 222. Conclusions TIMP3 is a novel target of miR -221 and miR -222 in glioma cell invasion.     

静滴复方氨基酸注射液致过敏性休克一例

患者女,22岁,农民。因上腹部饱胀半月,伴纳差、乏力、头昏来院。初诊:慢性胃炎。要求静滴复方氨基酸支持治疗,一次缓慢(15滴/min)静滴100ml,出现胸闷、全身搔痒、风疹团,四肢厥冷,继则上腹伴剧痛恶心呕吐,血压12/8kPa降至10/6kPa,心率120次/min,神志极度烦躁转为表情淡漠,立即停药,按抗过敏性休克处理,5小时后血压回升稳定于14.5/10kPa,48小时后皮疹消退痊愈出院。     

锁骨上神经加低位肌间沟臂丛神经阻滞术在锁骨开放内固定手术中的应用

目的观察锁骨上神经加低位肌间沟臂丛神经阻滞术在锁骨开放内固定术中麻醉效果。方法１２０例锁骨骨折病人被随机分为Ａ、Ｂ、Ｃ、Ｄ、Ｅ、Ｆ６组，每组均２０例，采用不同神经阻滞法均注入１．２％利多卡因＋０．３％布比卡因混合液（内含１：２０万肾上腺素），据患者主诉和骨科医师评定阻滞效果。结果Ｆ组中阻滞满意（Ⅰ、Ⅱ级）病人总例数明显多于Ａ、Ｂ、Ｃ、Ｄ、Ｅ５组（Ｐ＜０．０１），Ｆ组并发症发生率与其它各组比较无统计学差异。结论锁骨上神经加低位肌间沟臂丛神经阻滞术用于锁骨骨折手术病人，阻滞效果明显优于颈浅丛，臂丛（ｗｉｎｎｅ氏法及低位肌间沟法）及锁骨上神经阻滞。     

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.     

脑胶质瘤信号转导通路研究进展

脑胶质瘤是临床最为常见的原发性颅内肿瘤,约占颅内肿瘤的46%,占成年人全身肿瘤的2%;恶性胶质瘤的发病率为(5～8)/100万,在1998年     

靶向Ku70重组腺病毒shRNA在C6胶质瘤细胞的表达

目的 研究重组腺病毒Ad—Ku70shRNA对C6胶质瘤细胞系Ku70蛋白表达的影响，探讨放射治疗过程中基因增敏的新途径。方法以穿梭质粒为基础，构建重组腺病毒Ad—Ku70shRNA载体，扩增并鉴定其病毒滴度。然后转染至C6鼠脑胶质瘤细胞中，应用WestemBlot及免疫荧光观察其Ku70蛋白的表达情况，研究重组腺病毒Ad—Ku70shRNA对C6细胞Ku70表达的抑制效果。结果成功构建并扩增Ad—Ku70shRNA重组腺病毒，感染C6胶质瘤细胞后Ku70的表达明显降低。结论Ad—Ku70shRNA重组腺病毒载体可以明显降低C6胶质瘤细胞Ku70的表达。该结果为放射治疗基因增敏研究提供了有利的工具。     

反义miR-30a-5p抑制脑胶质瘤生长的体内实验研究

ObjectiveTo study the inhibitory effect of knocking down miR-30a-5p on the U87 human glioma xenograft growth and its possible mechanism. MethodsNude mice bearing subcutaneous U87 human glioblastoma were established and treated with miR-30a-5p antisense oligonucleotides (AS-miR-30a-5p) subcutaneous injection. Tumor size was measured every other day until the observation period ended. Researchers executed the animals after the treatment, stripped tumor tissues and extracted RNA and protein. Real-time PCR were conducted to detect the expression of miR-30a-5p. The histopathological characteristics and proliferation and apoptosis biological characters (including SEPT7, PCNA, cyclinD1, MMP-2, apoptosis related factor P53, bcl-2 and caspase3)were evaluated by HE and immunohistochemical staining, Western blot analysis respectively, and the cell apoptosis was detected by TUNEL method.ResultsIn AS-miR-30a-5p treated group, the tumor growth was delayed and the final tumor volume was smaller than that in the control and scr-ODN treated group (F=7.167, P＜0.05), and the expression of miR-30a-5p was knocked down. The expression of PCNA、cyclinD1 were significantly down-regulated while P53、SEPT7 and caspase3 up-regulated. Apoptotic index was increased significantly. ConclusionAs-miR-30a-5p suppresses the growth of U87 human gliomas xenografts significantly. Malignant phenotype of tumors are reversed to a considerable degree. Therefore, miR-30a-5p can be a candidate for targeted therapy of human glioma.     

Wnt5a在肿瘤研究中的进展

近年来研究发现WnL5a与肿瘤发生、发展关系密切.根据细胞类型不同,Wnt5a既可以起到促癌作用,又可以起到抑癌作用,其信号级联反应复杂,受多种因子调节.     

单纯放疗与宫颈植入性化疗协同放疗治疗老年宫颈癌的临床疗效对比分析

目的对比分析单纯放疗及应用植入性氟尿嘧啶同步放化治疗老年宫颈癌初治患者的疗效和并发症。方法选择2006年11月至2007年5月在吉林省肿瘤医院收治的初治宫颈癌患者共58例,临床分期为Ⅰb-Ⅲb期,按治疗方法不同分为单纯放疗组（共30例,给予X线远距离治疗机体外照射及192Ir腔内后装治疗）和同步放化疗组（共28例,给予宫颈植入氟尿嘧啶缓释剂500-800mg,同时给予X线远距离治疗机体外照射及192Ir腔内后装治疗）,对两组患者的疗效及并发症发生情况进行对比分析。结果单纯放疗组与同步放化疗组有效率分别为92%、86%,两组比较,差异有统计学意义（P〈0.05）两组患者近期并发症均以骨髓抑制为主,其中Ⅲ级以上骨髓抑发生率分别为3%、0%,两组比较,差异无统计学意义（P〉0.05）。结论宫颈植入性化疗协同放疗在不增加患者毒副反应的同时可明显改善老年宫颈癌患者的预后。     

生育及不育男性血清抑制素B水平的检测(简报)

不育男性激素评价报告表明,23％不育者伴有内分泌异常。而与男性生殖有关的主要内分泌腺是“下丘脑-垂体-睾丸轴系”,其内分泌调节是精子发生的重要环节,其中之一障碍或受其他疾病或因子干扰导致性腺激素生理水平紊乱均可能引发不育。因此,检测激素水平可评价下丘脑-垂体-