洛铂对结肠癌细胞的抗肿瘤活性及机制研究

Anti-tumor activity and mechanisms of lobaplatin in colon cancer cells

目的探索洛铂在结肠癌细胞中的抗肿瘤活性,阐明潜在分子机制,为临床应用提供理论证据。方法体外培养结肠癌细胞SW480,用MTT比色法和克隆形成实验检测不同浓度的洛铂分别处理结肠癌SW480细胞株24、48 h和72 h后,对结肠癌细胞增殖活性的抑制作用;用流式细胞仪检测不同浓度梯度的洛铂对结肠癌SW480细胞株凋亡率的影响;采用Western blot检测洛铂对SW480细胞凋亡相关蛋白CASPASE-3、BAD、BCL-2的表达;添加AKT抑制剂及激动剂,采用Western blot检测CASPASE3、AKT、p AKT、BAD、p-BAD和BCL-2表达量,探讨洛铂促结肠癌细胞凋亡的作用机制。结果洛铂能显著抑制体外培养的SW480细胞的生长,呈现为时间剂量依赖型;洛铂可诱导结肠癌SW480细胞凋亡,且随着洛铂浓度的增加,结肠癌SW480细胞凋亡率显著提高,结果具有统计学意义。洛铂可下调结肠癌细胞抑凋亡基因BCL-2蛋白表达,上调促凋亡基因BAD和CASPASE-3蛋白表达量;添加AKT抑制剂MK-2206后,pAKT、p-BAD和BCL-2蛋白表达显著降低,CASPASE3蛋白表达显著升高,促进了细胞凋亡,结果具有统计学差异。添加AKT激动剂SC97处理后,pAKT、p-BAD和BCL-2蛋白表达显著上调,CASPASE3蛋白表达显著下调,抑制了细胞凋亡,结果具有统计学意义。结论洛铂能显著抑制人结肠癌SW480细胞增殖,诱导癌细胞凋亡,洛铂可能通过AKT/BCL-2/BAD信号通路发挥促细胞凋亡作用,是一种有效的抗结肠癌药物。

Objective To explore the anti-tumor activity of lobaplatin in colon cancer cells,to elucidate the underlying molecular mechanisms,and to provide theoretical evidence for clinical application.Methods Colon cancer cell line SW480 was cultured in vitro.MTT colorimetric assay and colony formation assay were used to detect the inhibitory effect of different concentrations of lobaplatin on colon cancer SW480 cells at 24,48 and 72 h.The apoptosis rate of the cells was detected by flow cytometry.Western blot was used to detect the expression of the apoptosis-related proteins caspase-3,BAD and BCL-2 in SW480 cells after lobaplatin treatment.The mechanism of lobaplatin-induced apoptosis was investigated by adding AKT inhibitors and agonists and detecting the expression levels of caspase-3,AKT,pAKT,BAD,pBAD and BCL-2.Results Lobaplatin significantly inhibited the growth of cultured SW480 cells in a dose-and timedependent manner.Lobaplatin induced apoptosis of colon cancer SW480 cells.Lobaplatin induced downregulation of antiapoptotic BCL-2 protein expression and upregulated pro-apoptotic BAD and caspase-3 protein expression in the cells.Addition of AKT inhibitor MK-2206 significantly decreased p AKT,p-BAD,and BCL-2 expression,and significantly increased caspase-3 expression.Addition of AKT agonist SC97 significantly increased p AKT,p-BAD,and BCL-2 protein expression,and significantly decreased caspase-3 protein expression.Conclusions Lobaplatin significantly inhibited proliferation and induced apoptosis of human colon cancer SW480 cells.Lobaplatin may play a role in promoting apoptosis through the AKT/BCL-2/BAD signaling pathway,and it is an effective anti-colon cancer drug.