藤龙补中汤对大肠癌LoVo细胞失巢凋亡的作用及机制

Effects and mechanism of Tenglong Buzhong Decoction on anoikis of LoVo colorectal cancer cells

目的:观察藤龙补中汤对人大肠癌LoVo细胞失巢凋亡的作用及机制。方法:将藤龙补中汤作用于悬浮生长的LoVo细胞,CCK-8试剂检测细胞增殖;试剂盒检测失巢凋亡、Caspases活性和活性氧;Western blot检测蛋白表达和磷酸化。结果:藤龙补中汤可以抑制LoVo细胞悬浮生长,诱导失巢凋亡,活化Caspase-3、Caspase-8和Caspase-9。Caspases抑制剂Z-VAD-FMK可拮抗藤龙补中汤对失巢凋亡的作用。藤龙补中汤可以上调活性氧生成,活性氧清除剂N-乙酰-L-半胱氨酸可拮抗藤龙补中汤对Caspases的作用。藤龙补中汤还可降低整合素αV和β3的表达,抑制黏着斑激酶的磷酸化。结论:藤龙补中汤可诱导LoVo细胞失巢凋亡,其机制可能与上调活性氧、下调整合素αV和β3的表达、抑制黏着斑激酶的磷酸化相关。

Objective To study the effects and mechanism of Tenglong Buzhong Decoction(TBD)on anoikis in human colorectal cancer LoVo cells.Methods Suspension grown LoVo cells were treated with TBD.Cell proliferation was detected by cell counting kit-8(CCK-8)assay.Anoikis,Caspases activaties and reactive oxygen species(ROS)were measured by commercial kit.Proteins expression and phosphorylation were identified by Western blot.Results TBD inhibited suspension growth of LoVo cells,induced anoikis and activated Caspase-3,Caspase-8,and Caspase-9.Caspases inhibitor Z-VAD-FMK antagonized the effects of TBD on anoikis.TBD up-regulated ROS,ROS scavenger N-acetyl-L-cysteine(NAC)attenuated the effects of TBD on Caspases.TBD also inhibited the expressions of integrin aV andβ3,and phosphorylation of focal adhesion kinase(FAK).Conclusion TBD induces anoikis in LoVo cells,and may be related to up-regulation of ROS,and inhibition of integrin aV and integrinβ3 expressions,and FAK phosphorylation.