皖南蝮蛇毒抑瘤组分-Ⅰ对胃癌MKN-28细胞增殖、迁移及凋亡的影响

Effects of the proliferation,migration and apoptosis of AHVAC-Ⅰon gastric cancer MKN-28 cells

目的:探讨皖南蝮蛇毒抑瘤组分-Ⅰ(agkistrodon halys venom antitumor componentⅠ,AHVAC-Ⅰ)对胃癌MKN-28细胞生物学行为的影响作用。方法:不同实验浓度(5、10和15μg/mL)AHAVC-Ⅰ处理胃癌细胞MKN-2824 h后,采用细胞增殖和毒性检测(cell counting kit-8,CCK-8)法检测细胞增殖活力;划痕实验和Tanswell小室检测细胞迁移能力;激光共聚焦显微镜(annexin VmCherry/DAPI双染)和流式细胞术(annexin VFITC/PI双染)检测细胞凋亡;Western blot检测细胞凋亡相关蛋白Caspease-3的表达水平。结果:AHVAC-Ⅰ各浓度组分别与正常对照组相比结果显示,随着AHVAC-Ⅰ浓度的增加,MKN-28细胞增殖活力逐渐下降(P<0.01),细胞迁移能力逐渐被抑制(P<0.01),细胞凋亡率增高(P<0.05),凋亡相关蛋白Caspease-3表达上调(P<0.01)。结论:AHVAC-Ⅰ抑制胃癌细胞MKN-28增殖和迁移,诱导其凋亡。

AIM:To investigate the effects of agkis-trodon halys venom anti-tumor componentⅠ(AHVAC-Ⅰ)on the biological behavior of gastric cancer MKN-28 cells.METHODS:Gastric cancer MKN-28 cells were treated with the experimental concentrations(5,10,15μg/mL)of AHAVC-Ⅰfor 24 h.Cell proliferation and toxicity assay(cell counting kit-8,CCK-8)was used to detect the inhibition rates of the cells in different concentrations of AHVAC-Ⅰ.The migration ability of the cells was evaluated by wound-healing and Transwell assay.The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining,and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining.The protein level of Caspease-3 was determined by Western blot.RESULTS:Compared with normal control group,the results of AHVAC-Ⅰconcentration groups showed that with the increase of AHVAC-Ⅰconcentration,the proliferative activity of MN-28 cells decreased gradually(P<0.01),the cell migration ability decreased gradually(P<0.01),and the cell apoptosis rate increased(P<0.05).The expression of apoptosis-related protein Caspease-3 was up-regulated(P<0.01).CONCLUSION:AHVAC-Ⅰinhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.