下调RbAp48表达可抑制人宫颈癌MS751细胞株的迁移侵袭能力

目的探讨下调Rb Ap48表达对人宫颈癌细胞迁移侵袭的影响及其作用机制。方法 si RNA下调Rb Ap48表达,以划痕实验、Transwell小室检测下调Rb Ap48表达后对人宫颈癌细胞株MS751迁移侵袭的影响,Western bolt检测Vimentin、N-cadherin、E-cadherin、Snail、Twist、MMP-2、TIMP-2蛋白的表达。结果下调Rb Ap48表达后MS751细胞的迁移和侵袭数均明显减少(P<0.01);其间质细胞表型蛋白Vimentin、N-cadherin、MMP-2表达明显受到抑制;上皮细胞表型蛋白E-cadherin、TIMP-2表达明显升高,显示MS751细胞的上皮-间充质样变(EMT)受到抑制。Snail、Twist表达水平也发生显著下调。结论下调Rb Ap48表达能有效抑制宫颈癌MS751细胞株的上皮-间充质样变,降低细胞的迁移侵袭能力;其作用机制与降低Snail、Twist的表达有关。

重组腺病毒介导的RbAp48基因表达对人宫颈癌细胞生长、增殖的影响

目的构建携带RbAp48基因序列的重组腺病毒,检测其对宫颈癌细胞系SiHa肿瘤细胞行为的影响。方法构建穿梭质粒pDC316-RbAp48,与骨架质粒pBHGloxΔE1,3Cre共转染HEK293细胞,获得复制缺陷型重组腺病毒Ad5-RbAp48,Ad5-RbAp48感染人宫颈癌细胞SiHa,采用免疫印迹法检测RbAp48表达,使用WST-8、细胞计数及软琼脂克隆形成实验,研究Ad5-RbAp48介导RbAp48高表达对SiHa细胞增殖与非锚定依赖性生长的影响。结果成功构建并鉴定重组腺病毒Ad5-RbAp48,Ad5-RbAp48感染SiHa细胞导致RbAp48高效表达,并有效抑制SiHa细胞增殖能力与非锚定依赖性生长能力。结论 RbAp48重组腺病毒可介导RbAp48高效表达,具有抑制宫颈癌细胞肿瘤细胞行为的作用。

Effect of hypoxia and glutamine or glucose deprivation on the expression of retinoblastoma and retinoblastoma-related genes in ERN1 knockdown glioma U87 cell line

The expression of retinoblastoma and several retinoblastoma-related genes was studied in glioma cell line U87 and its subline with knockdown of ERN1 (endoplasmic reticulum—nuclei-1), the main endoplasmic reticulum stress sensing and signaling enzyme. It was shown that a blockade of the ERN1 enzyme function increases the expression levels of retinoblastoma, retinoblastoma-like 1 and most retinoblastoma related genes: EID1, JARID1B, E2F1, E2F3, RBAP48 and CTIP, does not change RNF40 and RBAP46 and decreases KDM5A. We have also demonstrated that hypoxia reduces the expression levels of retinoblastoma, EID1, and E2F1 in ERN1-deficient glioma cells only. At the same time, the expression levels of retinoblastoma-like 1, E2F3, RBAP46, RBAP48 and CTIP decrease, while JARID1B and RBBP2 increase in both types of cells in hypoxic conditions, but the expression is much stronger in cells with suppressed function of ERN1. The expression level of JARID1B and KDM-5A mRNA is also enhanced in glutamine deprivation condition in both tested cell types, moreover, this effect is amplified by the blockade of the ERN1 enzyme function. The expression levels of retinoblastoma, EID1, RBAP48, and E2F3 are decreased in glutamine deprivation condition only in ERN1-deficient glioma cells, but RBL1, CTIP, RBAP46, and E2F1—in both tested cell types with more significant effect in ERN1-deficient cells. Glucose deprivation condition leads to a decrease of expression levels of retinoblastoma, RBL1, E2F3, RBAP46, and RBAP48 in both used cell types and of EID1 and E2F1 only in glioma cells with suppressed function of signaling enzyme ERN1. Thus, expression levels of retinoblastoma and most retinoblastoma-related genes are increased under a blockade of ERN1 enzyme function and significantly changed in hypoxia, glucose or glutamine deprivation conditions both in control U87 cells and ERN1-deficient cells, but inhibition of the unfolded protein response sensor ERN1 predominantly enhances these effects. Moreover, it is possible that the induction of the expression of retinoblastoma and most retinoblastoma-related genes after knockdown of ERN1 plays an important role in suppression of glioma proliferation.

科学家发现造成老年性正常记忆减退的关键分子机制

2013年8月18日，因发现记忆的分子基础而荣获2000年诺贝尔医学奖的哥伦比亚大学EricKandel博士领衔的研究小组公布了一项最新研究成果，这项利用人脑和小鼠大脑进行的实验研究首次明确了造成老年性认知退化的记忆分子缺损机制。该研究小组在此项实验研究中，首先，从哥伦比亚大学纽约大脑库中采集年龄33～88岁无脑部疾病死者的8颗大脑，从中提取在大脑记忆功能中起中枢作用的海马部位齿状回（回飞镖形区域，其功能随年龄增长而下降，但不受阿尔茨海默病影响）和内嗅皮层（基本上不受年龄老化影响，但是最先受到阿尔茨海默病损伤即杀伤神经元的部位），