皖南蝮蛇毒抑瘤组分-Ⅰ逆转原代胃癌相关成纤维细胞促癌作用

AHVAC-Ⅰ reverses tumor growth of cancer-associated fibroblasats in gastric cancer

目的:探讨皖南蝮蛇毒抑瘤组分-Ⅰ(agkistrodon halys venom antitumor component-Ⅰ,AHVAC-Ⅰ)抑制胃癌细胞迁移的生物学活性。方法:组织块培养法和胰酶消化法分离培养人原代胃癌相关成纤维细胞(GCAFs);CCK8实验根据半数抑制浓度(IC 50)确定AHVAC-Ⅰ实验浓度;收集AHVAC-Ⅰ作用GCAFs后的上清液(GCAFs-CM^(AHVAC-Ⅰ)),划痕实验法和Transwell检测GCAFs-CM^(AHVAC-Ⅰ)抑制胃癌细胞株MKN-28迁移的能力;用ELISA、RT-PCR检测AHVAC-Ⅰ作用后GCAFs分泌产生的趋化因子CXCL12。结果:AHVAC-Ⅰ可显著抑制人原代GCAFs分泌的CXCL12(P<0.05),AHVAC-Ⅰ作用后生成的GCAFs-CM^(AHVAC-Ⅰ)明显抑制了胃癌细胞MKN-28的迁移能力(P<0.05)。结论:AHVAC-Ⅰ能够通过抑制GCAFs分泌生成的CXCL12抑制胃癌细胞MKN-28的迁移能力,逆转胃癌肿瘤微环境中的GCAFs的促进胃癌细胞转移的作用。

AIM: To explore whether Agkistrodon Halys venom antitumor component-Ⅰ(AHVAC-Ⅰ) affects the migration of gastric cancer cells by human primary gastric cancer-associated fibroblast(GCAFs). METHODS: Tissue block culture and trypsin digestion were used to separate and culture human primary gastric cancer-associated fibroblasts(GCAFs);the GCAFs-CM^(AHVAC-Ⅰ) was collected after AHVAC-Ⅰ treatment to culture the MKN28 cells. Migration was evaluated by wound-healing and Transwell assay. ELISA and RT-PCR were used to verify wether AHVAC-Ⅰ can play a role in the production of CXCL12 from GCAFs.RESULTS: AHVAC-Ⅰ can decrease the CXCL12 production from GCAFs(P<0.05). Cultured with GCAFs-CM^(AHVAC-Ⅰ), the migration of MKN28 cells were significantly inhibited(P<0.05). CONCLUSION: AHVAC-Ⅰ can inhibit the migration ability of gastric cancer cells MKN-28 by decreasing CXCL12 production from GCAFs, and is valuable for gastric cancer therapy.