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磷酸甘油酸激酶 1 促进 U251 人神经胶质瘤细胞的放射抗性。

Phosphoglycerate kinase 1 promotes radioresistance in U251 human glioma cells.

机构信息

Department of Neurosurgery, Nanjing Medical University, Affiliated Nanjing Brain Hospital, Nanjing, Jiangsu, P.R. China.

Neuro-Psychiatric Institute, Nanjing Medical University, Affiliated Nanjing Brain Hospital, Nanjing, Jiangsu, P.R. China.

出版信息

Oncol Rep. 2014 Feb;31(2):894-900. doi: 10.3892/or.2013.2874. Epub 2013 Nov 27.

Abstract

Phosphoglycerate kinase 1 (PGK1) has been found to be increased in radioresistant astrocytomas. The present study was designed to investigate the potential role of PGK1 in the radioresistance in U251 human cells. Quantitative PCR and western blot analysis were performed to evaluate PGK1 expression for mRNA levels and protein levels, respectively. The short hairpin RNA (shRNA)-PGK1 and the high expression plasmids were transfected to radioresistant U251 cells (RR-U251 cells) or normal U251 cells using lipofectamine™ 2000. The cell viability was determined by MTT assay. The wound-healing assay (WHA) was used to evaluate cell migration ability. Cell invasion abilities were examined using a Transwell culture chamber system. Our results showed that the expression of PGK1 was significantly increased in RR-U251 cells compared to normal U251 cells. Following irradiation, the cell viability as well as the migration and invasion ability were significantly higher in RR-U251 cells compared with normal U251 cells. Downregulating PGK1 using shRNA induced a significantly downregulated cell viability and decreased migration and invasion ability, and overexpression of PGK1 contributed to upregulated cell viability and increased migration and invasion ability, both in RR-U251 cells and normal U251 cells. These findings suggest that PGK1 could promote radioresistance in U251 human cells.

摘要

磷酸甘油酸激酶 1(PGK1)在耐辐射星形细胞瘤中被发现增加。本研究旨在探讨 PGK1 在 U251 人细胞中的放射抵抗作用中的潜在作用。定量 PCR 和 Western blot 分析分别用于评估 PGK1 的 mRNA 水平和蛋白水平的表达。短发夹 RNA(shRNA)-PGK1 和高表达质粒通过 lipofectamine™2000 转染至耐辐射 U251 细胞(RR-U251 细胞)或正常 U251 细胞。通过 MTT 测定法测定细胞活力。通过划痕愈合测定(WHA)评估细胞迁移能力。使用 Transwell 培养室系统检查细胞侵袭能力。我们的结果表明,RR-U251 细胞中 PGK1 的表达明显高于正常 U251 细胞。照射后,RR-U251 细胞的细胞活力以及迁移和侵袭能力明显高于正常 U251 细胞。使用 shRNA 下调 PGK1 可显著下调细胞活力,并降低迁移和侵袭能力,而过表达 PGK1 则有助于 RR-U251 细胞和正常 U251 细胞中细胞活力的上调和迁移及侵袭能力的增加。这些发现表明 PGK1 可促进 U251 人细胞的放射抵抗性。

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