Mechanism of overexpression of lncRNA LINC00152 in temozolomide-induced stem cell cycle arrest in glioma

张良龙; 安宏伟; 赵立智; 董　倩; 高桂艳

doi:10.19401/j.cnki.1007-3639.2020.05.002

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Mechanism of overexpression of lncRNA LINC00152 in temozolomide-induced stem cell cycle arrest in glioma

更新时间：2026-01-27

- Mechanism of overexpression of lncRNA LINC00152 in temozolomide-induced stem cell cycle arrest in glioma
- China Oncology Vol. 30, Issue 5, Pages: 328-334(2020)
- 作者机构：
  
  1. 沧州市人民医院神经外科,河北,沧州,061000
  2. 德州市陵城区中医院神经外科,山东,德州,253500
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2020.05.002
  CLC： R739.41
- Published Online：05 June 2020，
  
  Published：05 June 2020
- 稿件说明：
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张良龙 , 安宏伟 , 赵立智 , 董　倩 , 高桂艳 . LncRNA LINC00152过表达在替莫唑胺诱导脑胶质瘤干细胞周期阻滞中的作用机制探讨[J]. 中国癌症杂志, 2020, 30(5): 328-334.

张良龙, 安宏伟, 赵立智, et al. Mechanism of overexpression of lncRNA LINC00152 in temozolomide-induced stem cell cycle arrest in glioma[J]. China Oncology, 2020, 30(5): 328-334.
张良龙 , 安宏伟 , 赵立智 , 董　倩 , 高桂艳 . LncRNA LINC00152过表达在替莫唑胺诱导脑胶质瘤干细胞周期阻滞中的作用机制探讨[J]. 中国癌症杂志, 2020, 30(5): 328-334. DOI： 10.19401/j.cnki.1007-3639.2020.05.002.

张良龙, 安宏伟, 赵立智, et al. Mechanism of overexpression of lncRNA LINC00152 in temozolomide-induced stem cell cycle arrest in glioma[J]. China Oncology, 2020, 30(5): 328-334. DOI： 10.19401/j.cnki.1007-3639.2020.05.002.

摘要

背景与目的：LINC00152是异常表达于肝癌、胃癌及结肠癌等恶性肿瘤中的长链非编码RNA（long non-coding RNA，lncRNA）之一，参与增殖、迁移及凋亡等生物学行为，观察LINC00152在替莫唑胺（temozolomide，TMZ）诱导脑胶质瘤干细胞（glioblastoma stem cell，GSC）周期阻滞中的作用及其机制。方法：神经干细胞培养液培养脑胶质瘤U251细胞，并提取成球生长的GSC作为研究对象，慢病毒转染法外源性上调模型细胞LINC00152表达水平并以嘌呤霉素筛选稳定表达的细胞株，实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）检测LINC00152表达水平，细胞计数试剂盒（cell counting kit-8，CCK-8）法检测细胞活力变化，流式细胞术检测细胞周期变化，蛋白质印迹法（Western blot）检测P53、Cyclin A、CDC25A及CDK2蛋白水平变化。结果：分离提取的GSC呈球状生长，细胞球CD133和Oct-4呈阳性表达。慢病毒转染明显上调GSC中LINC00152表达水平（P＜0.000 1）。TMZ（200 μg·mL -1 ）处理48 h明显降低GSC活力（P＜0.000 1），而与空载对照组相比，LINC00152过表达能抵抗TMZ的诱导效应（P＜0.000 1）。TMZ处理48 h明显增加GSC的S期比例（P＜0.000 1），而与空载对照组相比，LINC00152过表达能抵抗TMZ的诱导效应（P＜0.000 1）。TMZ处理48 h明显下调GSC中Cyclin A、CDC25A、CDK2蛋白表达（P＜0.000 1），上调P53蛋白表达（P＜0.000 1）；而与空载对照组相比，LINC00152过表达能抵抗TMZ的诱导效应（P＜0.000 1）。结论：LncRNALINC00152过表达可抑制TMZ诱导的P53蛋白水平上调和CyclinA、CDC25A、CDK2蛋白水平下调，拮抗S期阻滞。

Abstract

Background and purpose: LINC00152 is a long non-coding RNA (lncRNA) that is abnormally expressed in malignant tumors such as liver cancer

gastric cancer and colon cancer. LINC00152 is involved in biological activities such as proliferation

migration and apoptosis. This study mainly investigated the role and mechanism of LINC00152 in temozolomide (TMZ)-induced gliosis of glioblastoma stem cell (GSC). Methods: Glioma U251 cells were cultured in the neural stem cell culture medium

and the GSC growing into the sphere were extracted as a research object. The expression level of the model cell LINC00152 was externally up-regulated using the lentiviral transfection method

and we screened the stably expressed cell line with puromycin. The expression level of LINC00152 was detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR). The cell viability was detected by cell counting kit-8 (CCK-8) method. The cell cycle was detected by flow cytometry. The expressions of P53

Cyclin A

CDC25A and CDK2 proteins were detected by Western blot. Results: The extracted GSC grew in a spherical shape

and the cell spheres CD133 and Oct-4 were positively expressed. Lentiviral transfection significantly up-regulated the expression level of LINC00152 in GSC (P0.000 1). The treatment of TMZ (200 μg·mL -1 ) for 48 h significantly reduced the viability of GSC (P0.000 1)

while the overexpression of LINC00152 was resistant to the induction effect of TMZ compared with the no-load control (P0.000 1). TMZ treatment for 48 h significantly increased the S phase ratio of GSC (P0.000 1)

while LINC00152 overexpression was resistant to TMZ induction compared with the empty control (P0.000 1). The expressions of Cyclin A

CDC25A and CDK2 in GSC were significantly down-regulated by TMZ for 48 h (P0.000 1)

while the expression of P53 protein was up-regulated (P0.000 1). Compared with the empty control

overexpression of LINC00152 was resistant to the induction of TMZ (P0.000 1). Conclusion: Overexpression of lncRNA LINC00152 inhibited TMZ-induced up-regulation of P53 protein expression and down-regulation of Cyclin A

CDC25A and CDK2 protein expressions

and antagonized S phase arrest of GSC.

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