华蟾素通过调控基质金属蛋白酶抑制人胃腺癌细胞MGC-803侵袭与迁移

倪腾洋; 李丹; 王海波; 陶丽; 金凤; 冯俊; 钱亚云; 陶立德; 砂川正隆; 刘延庆

doi:10.13422/j.cnki.syfjx.20181920

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华蟾素通过调控基质金属蛋白酶抑制人胃腺癌细胞MGC-803侵袭与迁移

中医药对胃癌的实验研究专题 | 更新时间：2024-01-04

- 华蟾素通过调控基质金属蛋白酶抑制人胃腺癌细胞MGC-803侵袭与迁移
- Effect of Cinobufacini on Invasion and Migration of Human Gastric Cancer MGC-803 Cells by Regulating Matrix Metalloproteinases
- 中国实验方剂学杂志 2018年24卷第19期页码：106-111
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81773944）;江苏省自然科学基金项目（BK20141280）
- DOI：10.13422/j.cnki.syfjx.20181920
  中图分类号： R285.5
- 纸质出版日期：2018
- 稿件说明：
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倪腾洋, 李丹, 王海波, 等. 华蟾素通过调控基质金属蛋白酶抑制人胃腺癌细胞MGC-803侵袭与迁移[J]. 中国实验方剂学杂志, 2018,24(19):106-111.

NI Teng-yang, LI Dan, WANG Hai-bo, et al. Effect of Cinobufacini on Invasion and Migration of Human Gastric Cancer MGC-803 Cells by Regulating Matrix Metalloproteinases[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 106-111.
倪腾洋, 李丹, 王海波, 等. 华蟾素通过调控基质金属蛋白酶抑制人胃腺癌细胞MGC-803侵袭与迁移[J]. 中国实验方剂学杂志, 2018,24(19):106-111. DOI： 10.13422/j.cnki.syfjx.20181920.

NI Teng-yang, LI Dan, WANG Hai-bo, et al. Effect of Cinobufacini on Invasion and Migration of Human Gastric Cancer MGC-803 Cells by Regulating Matrix Metalloproteinases[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 106-111. DOI： 10.13422/j.cnki.syfjx.20181920.

摘要

目的：探讨华蟾素对人胃腺癌MGC-803细胞的侵袭与迁移的影响及其分子机制。方法：噻唑蓝（MTT）比色法检测华蟾素（10，20，40，80，160，320 mg·L-1）对人胃腺癌MGC-803细胞增殖的抑制影响；transwell小室法检测华蟾素对MGC-803细胞侵袭与迁移能力的影响；蛋白免疫印迹法（Western blot）检测经华蟾素作用后的MGC-803细胞中基质金属蛋白酶-2（matrix metalloproteinases-2，MMP-2），基质金属蛋白酶-9（matrix metalloproteinases-9，MMP-9）和基质金属蛋白酶抑制因子-1（tissue inhibitors of metalloproteinases-1，TIMP-1），基质金属蛋白酶抑制因子-2（tissue inhibitors of metalloproteinases-2，TIMP-2）的蛋白表达水平；实时荧光定量PCR（Real-time PCR）检测经华蟾素处理后的MGC-803细胞中MMP-2，MMP-9，TIMP-1，TIMP-2的mRNA表达情况。结果：①MTT比色法结果显示，与空白组比较，华蟾素（10，20，40，80，160，320 mg·L-1）对胃腺癌MGC-803细胞的增殖具有显著抑制作用（P<0.01），并且华蟾素对MGC-803细胞的抑制作用呈浓度和时间依赖。②transwell实验结果显示，与空白组比较，华蟾素（10，20，40 mg·L-1）对胃腺癌MGC-803细胞的侵袭与迁移能力具有不同程度的抑制作用（P<0.05）③Real-time PCR和Western blot实验结果显示，与空白组比较，华蟾素（10，20，40 mg·L-1）能够下调MMP-2，MMP-9蛋白和mRNA的表达，上调TIMP-1，TIMP-2蛋白和mRNA的表达。结论：华蟾素能明显抑制胃腺癌MGC-803细胞的侵袭与迁移，其机制可能与上调TIMP-1，TIMP-2的转录水平，进而下调MMP-2，MMP-9蛋白水平有关。

Abstract

Objective: To investigate the effect of cinobufacini on the invasion and migration of human gastric adenocarcinoma MGC-803 cells and its molecular mechanisms. Method: The inhibitory effect of cinobufacini on the proliferation of human gastric cancer MGC-803 cells was detected by methye thiazolye telrazlium (MTT) assay. The effect of cinobufacini on the invasion and migration of MGC-803 cells was determined by transwell assay. The protein expressions of matrix metalloproteinases-2 (MMP-2)

matrix metalloproteinases-9 (MMP-9)

tissue inhibitors of metalloproteinases-1 (TIMP-1) and tissue inhibitors of metalloproteinases-2 (TIMP-2) were examined by Western blot. After treatment with different concentrations of cinobufacini

the mRNA expressions of MMP-2

MMP-9

TIMP-1 and TIMP-2 in MGC-803 cells were detected by Real-time PCR. Result: ① MTT assay showed that cinobufacini (10

160

320 mg·L-1) significantly inhibited the proliferation of gastric cancer MGC-803 cells (P<0.01) in a dose and time-dependent manner. ② Transwell assay results showed that cinobufacini (10

40 mg·L-1) inhibited the invasion and migration of gastric adenocarcinoma MGC-803 cells to varying degrees (P<0.05). ③ Real-time PCR and Western blot results showed that cinobufacini (10

40 mg·L-1) could down-regulate the protein and mRNA expressions of MMP-2

MMP-9 and up-regulate protein and mRNA expressions of TIMP-1

TIMP-2. Conclusion: Cinobufacini can obviously inhibit the invasion and migration of gastric cancer MGC-803 cells. The underlying mechanism may be related to the up-regulation of the transcription of TIMP-1 and TIMP-2 and the down-regulation of the protein levels of MMP-2 and MMP-9.

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