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二甲双胍对生物电场下肺癌H1975细胞增殖和定向迁移的影响
Metformin attenuates biological electric field-induced migration and proliferation of lung cancer cells by inhibiting AKT pathway
- 第三军医大学学报 2019年41卷第6期 页码：521-528
- 作者机构：
400042 重庆，陆军军医大学(第三军医大学)大坪医院呼吸内科
- 作者简介：
[何勇，E-mail：heyong8998@126.com HE Yong, E-mail:heyong8998@126.com
- 基金信息：国家自然科学基金面上项目(81672287);国家自然科学基金面上项目(81301840)
DOI：10.16016/j.1000-5404.201811011
中图分类号： R73-361; R734.2; R977.15收稿：2018-11-05，
修回：2019-01-08，
纸质出版：2019-03-30
- 稿件说明：
移动端阅览
胡晨, 李力, 卢从华, 等. 二甲双胍对生物电场下肺癌H1975细胞增殖和定向迁移的影响[J]. 第三军医大学学报, 2019,41(6):521-528.
Chen HU, Li LI, Conghua LU, et al. Metformin attenuates biological electric field-induced migration and proliferation of lung cancer cells by inhibiting AKT pathway[J]. Journal of Third Military Medical University, 2019, 41(6): 521-528.
胡晨, 李力, 卢从华, 等. 二甲双胍对生物电场下肺癌H1975细胞增殖和定向迁移的影响[J]. 第三军医大学学报, 2019,41(6):521-528. DOI： 10.16016/j.1000-5404.201811011.
Chen HU, Li LI, Conghua LU, et al. Metformin attenuates biological electric field-induced migration and proliferation of lung cancer cells by inhibiting AKT pathway[J]. Journal of Third Military Medical University, 2019, 41(6): 521-528. DOI： 10.16016/j.1000-5404.201811011.
摘要
目的
2
探讨二甲双胍对生物电场诱导的肺癌H1975细胞定向迁移和增殖的作用及其分子机制。
方法
2
利用外加直流电构建了生物电场诱导肺癌H1975细胞定向迁移的实验模型，观察不同电场梯度及实验分组下H1975细胞的迁移及增殖情况。实验分组：空白对照组、二甲双胍处理组(5 mmol/L二甲双胍预处理48 h)、电场处理组(100 mV/mm加电处理1 h)、二甲双胍联合电场处理组(5 mmol/L二甲双胍预处理48 h后，100 mV/mm加电处理1 h)。采用ImageXpress细胞成像微孔板检测仪采集肺癌H1975细胞运动图像，使用Image J软件分析细胞运动速度及运动定向性变化；Ki67荧光染色法检测细胞增殖能力；Western blot、免疫荧光法检测增殖和上皮-间质转化(epithelial-mesenchymal transition，EMT)相关蛋白的变化。
结果
2
在不同水平电场刺激作用下，H1975细胞的运动能力及定向性随着电场强度升高而不断增强(
P
<
0.05)，电场刺激还可增强细胞增殖能力并诱导EMT。在加入二甲双胍与电场联合处理后，细胞运动速度较电场处理组下降(
P
<
0.05)，趋电性减弱(
P
<
0.01)，细胞增殖能力受到抑制，同时E-cadherin表达水平较电场处理组升高(
P
<
0.01)、Vimentin表达水平降低(
P
<
0.05)，提示二甲双胍能够逆转电场诱导的EMT。Western blot结果显示电场刺激可上调p-AKT、p-ERK、p-STAT3的表达水平，而加入二甲双胍后可下调相关蛋白的表达。
结论
2
二甲双胍可通过抑制AKT/ERK信号通路及逆转EMT，抑制生物电场诱导的肺癌H1975细胞迁移和增殖。
Abstract
Objective
2
To observe the effect of metformin on migration and proliferation of H1975 cells induced by biological electric field and explore the possible mechanisms.
Methods
2
We established an experimental model to simulate biological electric field exposure that induced directional migration of lung cancer H1975 cells by constant direct current. H1975 cells were divided into blank control group
metformin treatment (for 48 h) group
EF group (with exposure to an electric field of 100 mV/mm for 1 h)
and combined treatment group (with metformin pretreatment 48 h before electric field exposure). Time-lapse images of the cells were acquired using the ImageXpress Micro high-throughput imager. The cell migration was quantified using ImageJ software with MTrackJ and Chemotaxis tool plugins. The effects of electric field exposure and metformin on the cell proliferation was examined using Ki67 incorporation assay. Immunofluorescence assay and Western blotting were performed to detect the expression of Ki67 and the markers of epithelial-mesenchymal transition (EMT) in the cells.
Results
2
H1975 cells migrated directionally to the cathode in the electric field in an intensity-dependent manner
while metformin significantly decreased both the directionality and the migration rate of the cells in the electric field. Electric field exposure also promoted the proliferation and induced EMT in H1975 cells
as evidenced by increased percentage of Ki67-positive cells
up-regulation of vimentin and down-regulation of E-cadherin in the cells. Metformin treatment significantly inhibited the proliferation and EMT of the cells either with or without electric field exposure. Western blotting showed that electric field exposure strongly increased the cellular expression of phosphorylated AKT
phosphorylated ERK
and phosphorylated STAT3
which were obviously suppressed by pretreatment of the cells with metformin.
Conclusion
2
Metformin can inhibit electric field-induced migration and proliferation of H1975 cells by inhibiting the AKT/ERK pathway and reversing EMT.
关键词
Keywords
references
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