α-红没药醇对胶质母细胞瘤细胞迁移和侵袭的影响

闫海斌; 徐如祥

doi:10.3969/j.issn.2095-5227.2018.08.015

α-红没药醇对胶质母细胞瘤细胞迁移和侵袭的影响

闫海斌^1,2,
徐如祥^2, ,

1. 解放军总医院, 北京 100853;
2. 陆军总医院附属八一脑科医院, 北京 100028

基金项目:

部委级资助项目；军事医学创新工程专项(16CXZ001)

详细信息

作者简介:
闫海斌,男,硕士,医师。研究方向:神经胶质瘤。Email:915227433@qq.com

通讯作者:
徐如祥,男,主任医师,博士生导师。Email:zjxuruxiang@163.com

中图分类号: R 739.41
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出版历程
- 收稿日期: 2018-04-24
- 修回日期: 2018-04-24
- 网络出版日期: 2023-11-25

Effect of α-bisabolol on migration and invasion of glioblastoma cells

YAN Haibin^1,2,
XU Ruxiang^2, ,

1. Chinese PLA General Hospital, Beijing 100853, China;
2. Bayi Brain Hospital Affiliated to PLA Army General Hospital, Beijing 100028, China

Funds:

Supported by the Military Medical Innovation Project Special (16CXZ001)

摘要

摘要: 目的研究α-红没药醇(α-bisabolol)对胶质母细胞瘤细胞U251和U87迁移和侵袭的影响并探讨其可能的机制。方法体外培养人脑胶质母细胞瘤细胞U251和U87，CCK8比色法检测不同浓度(0μmol/L、1.25μmol/L、2.5μmol/L、5μmol/L、10μmol/L)α-红没药醇作用24 h后细胞存活率的变化；细胞划痕实验、Transwell实验、Western blotting检测不同浓度(0μmol/L、1.25μmol/L、2.5μmol/L)α-红没药醇对U251和U87细胞迁移、侵袭能力及对细胞MMP-2、MMP-9、c-Met蛋白表达量的影响；U251细胞分为A组(空载质粒组)、B组(空载质粒+2.5μmol/Lα-红没药醇组)、C组(过表达c-Met质粒组)、D组(过表达c-Met质粒+2.5μmol/Lα-红没药醇组)，转染质粒后24 h，B、D组加入2.5μmol/Lα-红没药醇，细胞划痕实验、Transwell实验、Western blotting分别检测4组细胞迁移、侵袭能力及c-Met、MMP-2、MMP-9蛋白表达量。结果 5μmol/L、10μmol/L组细胞存活率较0μmol/L、1.25μmol/L、2.5μmol/L组下降(U251：61.22%±5.08%、29.48%±4.84% vs 100%±0.00%、98.16%±5.71%、96.89±7.30%，P=0.00；U87：55.72%±8.17%、19.66%±4.82% vs 100%±0.00%、97.86%±5.41%、95.31%±5.42%，P=0.00)。0μmol/L、1.25μmol/L、2.5μmol/L组划痕愈合百分比分别为U251：49.36%±5.44%、35.08%±3.79%、23.89%±4.51%，U87：46.64%±4.83%、33.42%±3.10%、22.35%±3.62%，侵袭到下室的细胞数量分别为U251：248.67±14.94、171.11±17.91、87.11±15.49，U87：202.44±16.98、145.44±11.91、71.98±9.32，三组间差异均有统计学意义(P均=0.000)。c-Met、MMP-2、MMP-9蛋白表达量分别为：U251(c-Met：1.00±0.00、0.70±0.09、0.33±0.08；MMP-2：1.00±0.00、0.70±0.08、0.32±0.10；MMP-9：1.00±0.00、0.69±0.09、0.24±0.07)；U87(c-Met：1.00±0.00、0.71±0.08、0.27±0.08；MMP-2：1.00±0.00、0.71±0.10、0.29±0.04；MMP-9：1.00±0.00、0.72±0.08、0.23±0.04)，三组间差异均有统计学意义(P=0.000)。转染c-Met过表达质粒后D组较B组划痕愈合百分比明显升高(35.61%±4.70% vs 13.11%±2.99%)，D组侵袭到下室的细胞数量较B组明显升高(209.44±18.13 vs 91.33±14.46)，D组较B组c-Met、MMP-2、MMP-9蛋白表达量明显升高(c-Met：0.60±0.04 vs 0.39±0.04；MMP-2：0.71±0.10 vs 0.49±0.08，MMP-9：0.73±0.11 vs 0.45±0.07，P=0.000)。结论 α-红没药醇可以通过下调c-Met抑制胶质母细胞瘤细胞迁移和侵袭。
- α-红没药醇 /
- 胶质母细胞瘤 /
- 迁移 /
- 侵袭 /
- c-Met
Abstract: Objective To study the effect of α-bisabolol on migration and invasion of glioblastoma cells and explore its possible mechanism. Methods Human glioblastoma cell lines U251 and U87 were cultured in vitro. CCK8 colorimetric assay was used to detect the changes of cell viability at 24 hours after treatment with different concentrations of α-bisabolol (0 µmol/L, 1.25 µmol/L, 2.5 µmol/L, 5 µmol/L, 10 µmol/L); Cell scratch test, Transwell assay and Western blotting were used to detect the changes of cell migration and invasive ability and changes in the expression of MMP-2, MMP-9 and c-Met protein at 24 hours after treatment with different concentrations of α-bisabolol (0 µmol/L, 1.25 µmol/L, 2.5 µmol/L); U251 cells were divided into four groups: group A(blank plasmid), group B (blank plasmid +2.5 µmol/L α-bisabolol), group C (c-Met over-expressing plasmid) and group D (c-Met over-expressing plasmid +2.5 µmol/L α-bisabolol). At 24 hours after plasmid transfection, 2.5 µmol/L of α-bisabolol was added to group B and D, Cell scratch test, Transwell assay and Western blotting were used to detect cell migration, invasion and expression of c-Met, MMP-2, and MMP-9 proteins in the four groups. Results The cell survival rate of 5 µmol/L, 10μmol/L group was lower than that of 0 µmol/L, 1.25 µmol/L, 2.5μmol/L group (U251: 61.22%±5.08%, 29.48%±4.84% vs 100%±0.00%, 98.16%±5.71%, 96.89±7.30%, P=0.00, U87: 55.72%±8.17%, 19.66%±4.82% vs 100%±0.00%, 97.86%±5.41%, 95.31%±5.42%, P=0.00).The percentage of scratch closure in the 0μmol/L, 1.25μmol/L and 2.5μmol/L groups was 49.36%±5.44%, 35.08%±3.79%, 23.89%±4.51% for U251, and 46.64%±4.83%, 33.42%±3.10%, 22.35%±3.62% for U87. The number of cells invaded the lower chamber was 248.67±14.94, 171.11±17.91, 87.11 ±15.49 for U251, and 202.44±16.98, 145.44±11.91, 71.98±9.32 for U87, the differences between three groups were statistically significant (all P=0.000). c-Met, MMP-2, and MMP-9 protein expression levels were: U251 (c-Met: 1.00±0.00, 0.70±0.09, 0.33±0.08, MMP-2: 1.00±0.00, 0.70±0.08, 0.32±0.10, MMP-9: 1.00±0.00, 0.69±0.09, 0.24±0.07), U87 (c-Met: 1.00±0.00, 0.71±0.08, 0.27±0.08, MMP-2: 1.00±0.00, 0.71±0.10, 0.29±0.04, MMP-9:1.00±0.00, 0.72±0.08, 0.23±0.04), the differences between three groups were statistically significant (all P=0.000). After transfection of c-Met overexpression plasmid, the percentage of scratch closure in group D was significantly higher than that in group B (35.61%±4.70% vs 13.11%±2.99%, P=0.00). The number of cells invaded the lower chamber in group D was significantly higher than that in group B (209.44±18.13 vs 91.33±14.46, P=0.00). The expression of c-Met, MMP-2, and MMP-9 in group D was higher than that in group B (c-met: 0.60±0.04 vs 0.39±0.04; MMP-2: 0.71±0.10 vs 0.49±0.08; MMP-9: 0.73±0.11 vs 0.45±0.07), and the differences were statistically significant (all P=0.000). Conclusion α-Bisabolol can inhibit migration and invasion of glioblastoma cells by down-regulating c-Met.
- α-bisabolol /
- glioblastoma /
- migration /
- invasion /
- c-Met