解剖学报

解剖学报 ›› 2019, Vol. 50 ›› Issue (5): 595-600.doi: 10.16098/j.issn.0529-1356.2019.05.009

• 肿瘤生物学 • 上一篇    下一篇

曲古抑菌素A通过上调原钙黏蛋白9表达抑制人胃癌SGC-7901细胞迁移和侵袭

程琦1 刘晓东2 徐宛玲3*    

  1. 1.漯河医学高等专科学校病理学教研室; 2.漯河医学高等专科学校解剖学教研室; 3.内科学教研室，河南 漯河 462002
  • 收稿日期:2018-08-15 修回日期:2018-10-15 出版日期:2019-10-06 发布日期:2019-12-10
  • 通讯作者: 徐宛玲 E-mail:872189991@qq.com
  • 基金资助:
    河南省科技厅科技发展计划项目

Tichostatin A inhibiting migration and invasion of human gastric carcinoma SGC-7901 cells via up-regulating protocadherin 9

CHENG Qi1 LIU Xiao-dong2 XU Wan-ling 3*   

  1. 1.Department of Pathology; 2.Department of Anatomy; 3.Department of Internal Medicine, Luohe Medical College, He’nan Luohe 462002, China
  • Received:2018-08-15 Revised:2018-10-15 Online:2019-10-06 Published:2019-12-10
  • Contact: Wan-ling XU E-mail:872189991@qq.com

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摘要：

目的 探讨曲古抑菌素A（TSA）对人胃癌SGC-7901细胞迁移和侵袭能力的影响，及其可能的分子机制。 方法 体外常规培养SGC-7901细胞至对数生长期，给予不同浓度TSA（5、10、20、40、80、160 nmol/L）处理48 h，采用细胞计数试剂盒（CCK-8）法检测细胞活力，脂质体介导法将人原钙黏蛋白9（PCDH9）真核表达质粒转染SGC-7901细胞并筛选出稳定表达的细胞系，Transwell实验检测迁移率和侵袭率，RT-PCR法检测PCDH9 mRNA表达水平，免疫印迹法(Western blotting)检测PCDH9、Snail、E-钙黏蛋白（E-cadherin）、基质金属蛋白酶2（MMP-2）和MMP-9的蛋白表达水平。 结果 TSA在80 nmol/L以上时对人胃癌SGC-7901细胞活力有显著的抑制作用（P<0.05）；而在较低浓度下（5～20 nmol/L）可剂量依赖性地抑制人胃癌SGC-7901细胞的迁移和侵袭能力（P<0.05），下调Snail、MMP-2和MMP-9蛋白表达水平（P<0.05），上调E-cadherin和PCDH9蛋白表达水平（P<0.05），上调PCDH9 mRNA表达水平（P<0.05）；PCDH9高表达也可抑制SGC-7901细胞迁移和侵袭（P<0.05），上调E-cadherin蛋白表达并下调Snail、MMP-2和MMP-9蛋白表达（P<0.05）。 结论 TSA可在体外抑制胃癌SGC-7901细胞迁移和侵袭，这可能与其促进PCDH9基因表达，继而下调Snail、MMP-2和MMP-9蛋白表达，上调E-cadherin蛋白表达有关。

关键词: 曲古抑菌素A, 原钙黏蛋白9, 胃癌, SGC-7901细胞, 免疫印迹法, 人

Abstract:

Objective To investigate the effect of trichostatin A (TSA) on migration and invasion in human gastric carcinoma SGC-7901 cells and its possible mechanism. Methods SGC-7901 cells were cultured in vitro and treated with TSA (5, 10, 20, 40, 80, 160 nmol/L) for 48 hours, and then the cell viability was detected by cell counting kit 8(CCK-8) assay. The protocadherin 9 (PCDH9) high-expression SGC-7901 cells were stably established by transfecting with eukaryotic expression vector (pCMV6-PCDH9). Transwell assay was used to determine the abilities of migration and invasion. The mRNA expression level of PCDH9 were measured by RT-PCR. Western blotting was performed to analyze the protein expression of PCDH9, Snail, E-cadherin, matrix metalloproteinase (MMP)-2 and MMP-9. Results TSA remarkably reduced the cell viability of SGC-7901 cells in excess of 80 nmol/L (P<0.05). However, in a dose-dependent manner, low-level TSA (5-20 nmol/L) suppressed migration and invasion of SGC-7901 cells (P<0.05), down-regulated the protein levels of Snail, MMP-2 and MMP-9 (P<0.05), and up-regulated the protein levels of PCDH9 and E-cadherin (P<0.05). Meanwhile, high expression of PCDH9 also inhibited migration and invasion of SGC-7901 cells (P<0.05), down-regulated the protein levels of Snail, MMP-2 and MMP-9 (P<0.05), and up-regulated the protein level of E-cadherin (P<0.05). Conclusion TSA may inhibit migration and invasion of SGC-7901 cells most likely via up-regulating PCDH9, and then down-regulating the protein levels of Snail, MMP-2 and MMP-9, and up-regulating the protein level of E-cadherin.

Key words: Trichostatin A, Protocadherin 9, Gastric carcinoma, SGC-7901 cell, Western blotting, Human