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

doi:10.19401/j.cnki.1007-3639.2020.05.002

中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (5): 328-334.doi: 10.19401/j.cnki.1007-3639.2020.05.002

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

张良龙¹ ，安宏伟 ² ，赵立智 ¹ ，董　倩 ¹，高桂艳¹

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

出版日期:2020-05-30 发布日期:2020-06-05
通信作者: 张良龙 E-mail: zhangweihospital@sina.cn
基金资助:
2018年沧州市科技计划项目（183302129）；2016年沧州市科技计划项目（162302072）。

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

ZHANG Lianglong ¹, AN Hongwei ² , ZHAO Lizhi ¹ , DONG Qian ¹, GAO Guiyan ¹

1. Department of Neurosurgery, Cangzhou People’s Hospital, Cangzhou 061000, Hebei Province, China; 2. Department of Neurosurgery, Dezhou Lingcheng Hospital of Traditional Chinese Medicine, Dezhou 253500, Shandong Province, China

Published:2020-05-30 Online:2020-06-05
Contact: ZHANG Lianglong E-mail: zhangweihospital@sina.cn

摘要/Abstract

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。

关键词: 长链非编码RNA, LINC00152, 替莫唑胺, 细胞周期, 脑胶质瘤

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 (P<0.000 1). The treatment of TMZ (200 μg·mL -1 ) for 48 h significantly reduced the viability of GSC (P<0.000 1), while the overexpression of LINC00152 was resistant to the induction effect of TMZ compared with the no-load control (P<0.000 1). TMZ treatment for 48 h significantly increased the S phase ratio of GSC (P<0.000 1), while LINC00152 overexpression was resistant to TMZ induction compared with the empty control (P<0.000 1). The expressions of Cyclin A, CDC25A and CDK2 in GSC were significantly down-regulated by TMZ for 48 h (P<0.000 1), while the expression of P53 protein was up-regulated (P<0.000 1). Compared with the empty control, overexpression of LINC00152 was resistant to the induction of TMZ (P<0.000 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.

Key words: Long non-coding RNA, LINC00152, Temozolomide, Cell cycle, Glioma

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.

ZHANG Lianglong , AN Hongwei , ZHAO Lizhi , DONG Qian , GAO Guiyan . Mechanism of overexpression of lncRNA LINC00152 in temozolomide-induced stem cell cycle arrest in glioma[J]. China Oncology, 2020, 30(5): 328-334.

[1]	金优萍, 李录英, 周　平. 长链非编码RNA STMN1P2在乳腺癌中的作用及机制研究[J]. 中国癌症杂志, 2021, 31(7): 596-604.
[2]	高　超 , 田彦明 , 丁博月 , 张　翼 . Musashi1基因调控结肠癌HCT116细胞恶性生物学行为的机制研究[J]. 中国癌症杂志, 2020, 30(3): 199-207.
[3]	王晓松, 陈俊霞. 环状RNA hsa_circ_0050900在乳腺癌中的表达及其对细胞生物学行为影响的机制研究[J]. 中国癌症杂志, 2020, 30(12): 977-983.
[4]	朱秀之, 陈　力, 纪　磊, 高　雨, 王中华. CDK4/6抑制剂治疗三阴性乳腺癌的研究进展[J]. 中国癌症杂志, 2019, 29(11): 899-905.
[5]	杨　睿,陈俊霞. 环状RNA hsa_circ_0058514在三阴性乳腺癌中的表达及作用研究[J]. 中国癌症杂志, 2019, 29(1): 9-18.
[6]	张　雷,刘一博,刘继才,等. 评价替莫唑胺对脑胶质瘤治疗敏感性的新方法[J]. 中国癌症杂志, 2018, 28(9): 657-664.
[7]	王秀月,赵　川,陈　彻,等. 长链非编码RNA作为肝细胞癌诊断新型血清标志物的meta分析[J]. 中国癌症杂志, 2018, 28(3): 229-235.
[8]	许文芳,费迎明,周建康,等. XB130基因对肝细胞癌细胞增殖的影响及其机制[J]. 中国癌症杂志, 2018, 28(2): 117-122.
[9]	池虹,陈俊霞. LncRNA RP11-79H23.3在膀胱癌细胞中的作用及其发生、发展的研究[J]. 中国癌症杂志, 2018, 28(1): 22-29.
[10]	刘　磊,吴　炅. 乳腺癌相关长链非编码RNA的研究进展[J]. 中国癌症杂志, 2017, 27(8): 668-674.
[11]	刘　宁,成冬冬,姜金波. 长链非编码RNA PANDAR促进结直肠癌转移的作用和机制研究[J]. 中国癌症杂志, 2017, 27(4): 268-275.
[12]	李妮娅,左雪梅,李　莉,等. 斯钙素1的表达对肺癌细胞A549细胞周期及凋亡的影响[J]. 中国癌症杂志, 2016, 26(8): 641-647.
[13]	何安邦,陈浩冬,吕兆洁,等. 肾癌相关长链非编码RNA研究进展[J]. 中国癌症杂志, 2016, 26(8): 704-711.
[14]	朱竹先,余　晨,邱忠民,等. LncRNA PCGEM1和雄激素受体在前列腺癌中的共定位表达及临床意义[J]. 中国癌症杂志, 2016, 26(4): 320-325.
[15]	朱一平,叶定伟. 长链非编码RNA在泌尿系统肿瘤中的研究进展[J]. 中国癌症杂志, 2016, 26(1): 117-120.

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

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价