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14-3-3-β在人恶性胶质瘤细胞系U87MG迁移和侵袭中的作用

Role of 14-3-3-beta in the migration and invasion in human malignant glioma cell line U87MG.

作者信息

Park Sung-Geun, Jung Shin, Ryu Hyang-Hwa, Jung Tae-Young, Moon Kyung-Sub, Kim In-Young, Jeong Young-Il, Pei Jian, Park Seung-Jin, Kang Sam-Suk

机构信息

Department of Neurosurgery, Chonnam National University Hwasun Hospital and Medical School, Gwangju, Korea.

出版信息

Neurol Res. 2012 Nov;34(9):893-900. doi: 10.1179/1743132812Y.0000000087. Epub 2012 Aug 22.

Abstract

PURPOSE

To assess the influence of 14-3-3-beta in modulating the migration and invasion of human glioma cells.

METHODS

To profile the genes associated with malignant glioma cell motility, differential display-polymerase chain reaction was performed and the findings were validated by Northern blotting in the U343MG-A, U87MG, and U87MG-10' human glioma cell lines. Antisense 14-3-3-beta cDNA plasmid was transfected into U87MG ('U87-YA-3'). To follow motility changes after transfection, simple scratch test and matrigel assay were performed. Morphological and cytoskeletal changes were documented by light and confocal microscopy. In addition, doubling times of the transfectant and endogenous 14-3-3-beta levels were determined in various glioma cell lines with different motilities.

RESULTS

14-3-3-beta was highly expressed in U87MG cells. U87-YA-3 cells became small and flat, and actin was depolarized. Furthermore, U87-YA-3 cell motility was inhibited markedly versus parental U87MG cells. The doubling times of transfected and parent cells were 32 and 37 hours, respectively. Endogenous 14-3-3-beta expression in the human glioma cell lines was proportional to their migratory and invasive abilities.

CONCLUSION

14-3-3-beta modulates the migration and invasion in U87MG cells, which may be useful in developing therapeutic approaches for the treatment of glioma.

摘要

目的

评估14-3-3-β在调节人胶质瘤细胞迁移和侵袭中的作用。

方法

为了分析与恶性胶质瘤细胞运动相关的基因,采用差异显示聚合酶链反应,并通过Northern印迹法在U343MG-A、U87MG和U87MG-10'人胶质瘤细胞系中验证结果。将反义14-3-3-β cDNA质粒转染到U87MG细胞中(“U87-YA-3”)。为了跟踪转染后的运动变化,进行了简单划痕试验和基质胶试验。通过光学显微镜和共聚焦显微镜记录形态学和细胞骨架的变化。此外,还测定了不同运动能力的各种胶质瘤细胞系中转染细胞的倍增时间和内源性14-3-3-β水平。

结果

14-3-3-β在U87MG细胞中高表达。U87-YA-3细胞变得小而扁平,肌动蛋白去极化。此外,与亲本U87MG细胞相比,U87-YA-3细胞的运动能力明显受到抑制。转染细胞和亲本细胞的倍增时间分别为32小时和37小时。人胶质瘤细胞系中内源性14-3-3-β的表达与其迁移和侵袭能力成正比。

结论

14-3-3-β调节U87MG细胞的迁移和侵袭,这可能有助于开发治疗胶质瘤的方法。

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