谷氨酸受体拮抗剂对恶性黑素瘤WM451LU细胞增殖、迁移的调控作用及相关机制研究

中华皮肤科杂志 ›› 2016, Vol. 49 ›› Issue (8): 578-581.

谷氨酸受体拮抗剂对恶性黑素瘤WM451LU细胞增殖、迁移的调控作用及相关机制研究

李丽丽¹,陈显峰²,黄琦涛³,潘南楠¹,徐文英¹,谢治³

1. 南宁广西壮族自治区人民医院皮肤科
2. 广西医科大学第一附属医院重症医学科
3. 广西自治区人民医院皮肤科

收稿日期:2015-09-09 修回日期:2016-04-15 发布日期:2016-08-05
通讯作者: 谢治 E-mail:xiezhi11@aliyun.com

Regulatory effects of glutamate receptor antagonists on the proliferation and migration of WM451LU malignant melanoma cells and their related mechanisms

Received:2015-09-09 Revised:2016-04-15 Published:2016-08-05

摘要/Abstract

摘要：

目的探讨谷氨酸受体拮抗剂对恶性黑素瘤WM451LU细胞增殖、迁移的调控作用及相关机制。方法取对数生长期人侵袭性恶性黑素瘤WM451LU细胞，分为3组，分别接受100 μmol/L谷氨酸受体拮抗剂MK?801（MK?801组）、10 μmol/L谷氨酸受体拮抗剂CPCCOEt（CPCCOEt组）或单纯培养基（对照组）处理。24 h后，噻唑蓝（MTT）法检测细胞增殖率，划痕实验检测细胞迁移能力，免疫印迹法检测增殖细胞核抗原（PCNA）、细胞膜及胞质中蛋白激酶Cα（PKCα）以及磷酸化MAPK（p?MAPK）表达水平。结果处理24 h后，对照组、MK?801组以及CPCCOEt组细胞增殖率分别为100% ± 1.1%、63% ± 3.1%、60% ± 2.4%，后两组与对照组比较，差异均有统计学意义（P < 0.05）。划痕实验结果显示，对照组无细胞带随着时间的推移而逐渐变窄，划痕趋于愈合状态，而经MK?801及CPCCOEt作用后，无细胞带的变窄速度要明显缓慢，培养24 h后无细胞带仍然较宽，缩窄程度不明显。MK?801及CPCCOEt组细胞PCNA蛋白表达水平分别为77.0% ± 5.4%和72.0% ± 4.2%，显著低于对照组（100.0% ± 1.3%），差异均有统计学意义（P < 0.05）。对照组、MK?801组以及CPCCOEt组细胞膜上PKCα表达水平分别为0.38 ± 0.01、0.12 ± 0.02、0.14 ± 0.02，后两组与对照组比较，差异均有统计学意义（P < 0.05）。MK?801组及CPCCOEt组p?MAPK表达水平分别为0.48 ± 0.03、0.36 ± 0.04，显著低于对照组（1.00 ± 0.02），差异均有统计学意义（P < 0.05）。结论体外阻滞谷氨酸受体能够抑制WM451LU细胞增殖、迁移，该作用可能部分由PKCα?MAPK信号通路介导。

Abstract:

Li Lili, Chen Xianfeng, Huang Qitao, Pan Nannan, Xu Wenying, Xie Zhi Department of Dermatology, People′s Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China （Li LL, Huang QT, Pan NN, Xu WY, Xie Z）; Department of Critical Care Medicine, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China Corresponding author: Xie Zhi, Email: xiezhi11@aliyun.com 【Abstract】 Objective To evaluate regulatory effects of glutamate receptor antagonists on the proliferation and migration of WM451LU malignant melanoma cells, and to explore their related mechanisms. Methods WM451LU cells at exponential growth phase were classified into 3 groups to be treated with the glutamate receptor antagonist MK?801 at 100 μmol/L （MK?801 group）, the glutamate receptor antagonist CPCCOEt at 10 μmol/L （CPCCOEt group）, or culture medium （control group）. After 24?hour treatment, methyl thiazolyl tetrazolium （MTT）assay was performed to determine cell proliferation rates, scratch assay to evaluate the migration activity of cells, and Western?blot analysis to measure levels of proliferating cell nuclear antigen （PCNA）, protein kinase Cα （PKCα） both on cell membrane and in cytoplasm, and phosphorylated mitogen?activated protein kinase （p?MAPK）. Results After 24?hour treatment, cell proliferation rates were significantly decreased in the MK?801 group and CPCCOEt group compared with the control group （63% ± 3.1% and 60% ± 2.4% vs. 100% ± 1.1%, both P < 0.05）. The scratch assay showed that cell?free zones in the control group gradually narrowed over time, and the scratch wound tended to close. However, the cell?free zones in the MK?801 group and CPCCOEt group narrowed more slowly compared with the control group, and were still wide after 24?hour culture with no obvious closure of the scratch. The MK?801 group and CPCCOEt group both showed significantly decreased s of PCNA （77.0% ± 5.4% and 72.0% ± 4.2% respectively）, PKCα on the cell membrane （0.12 ± 0.02 and 0.14 ± 0.02 respectively）, and p?MAPK （0.48 ± 0.03 and 0.36 ± 0.04 respectively） compared with the control group （PCNA: 100.0% ± 1.3%; PKCα: 0.38 ± 0.01; p?MAPK: 1.00 ± 0.02; all P < 0.05）. Conclusion In vitro suppression of glutamate receptors can inhibit the proliferation and migration of WM451LU cells, likely through the mediation of the PKCα?MAPK signaling pathway.

中图分类号:

R758.5

李丽丽陈显峰黄琦涛潘南楠徐文英谢治. 谷氨酸受体拮抗剂对恶性黑素瘤WM451LU细胞增殖、迁移的调控作用及相关机制研究[J]. 中华皮肤科杂志, 2016,49(8):578-581. doi:

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