木香烃内酯通过PI3K/AKT信号通路调控结直肠癌细胞增殖、凋亡、侵袭和迁移

The Regulation of Costunolide on Colorectal Cancer Cell Proliferation,Apoptosis,Invasion and Migration through PI3K/AKT Signaling Pathway

目的:探讨木香烃内酯对结直肠癌细胞增殖、凋亡、侵袭和迁移的影响及作用机制。方法:不同浓度(10、20、40μmol/L)木香烃内酯作用结直肠癌细胞SW480,并设空白对照组、溶剂对照组和紫杉醇组,MTT和平板克隆实验检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell检测细胞迁移和侵袭,Western Blot检测与增殖、凋亡、侵袭和迁移相关蛋白、LASP1、PI3K/AKT信号通路相关蛋白表达。应用LipofectamineTM2000将真核表达载体pCDNA-LASP1和空载体pCDNA导入SW480细胞,观察LASP1过表达后木香烃内酯对SW480增殖、凋亡、迁移和侵袭及PI3K/AKT信号通路的影响。结果:与空白对照组比较,40μmol/L木香烃内酯组SW480细胞活力、细胞迁移和侵袭数量显著降低、凋亡率显著升高,CDK1、CyclinD1、MMP-2、MMP-9、LASP1、PI3K、AKT、mTOR蛋白表达显著降低,Caspase-3、Caspase-9蛋白表达显著升高。空白对照组和溶剂对照组各检测指标差异无统计学意义(P>0.05)。LASP1过表达可逆转木香烃内酯对SW480细胞增殖、迁移、侵袭及PI3K、AKT、mTOR蛋白表达的抑制作用,并逆转木香烃内酯对SW480细胞的凋亡促进作用。结论:木香烃内酯可能通过抑制PI3K/AKT信号通路调控结直肠癌细胞的增殖、凋亡、侵袭和迁移。

Objective:To investigate the effects of costunolide on proliferation,apoptosis,invasion and migration of colorectal cancer cells and its mechanism.Methods:Colorectal cancer cells SW480 were treated with costunolide at different concentrations(10,20,40μmol/L),blank control group,solvent control group and paclitaxel group were set up.MTT and plate cloning assays were used to detect cell proliferation,flow cytometry was used to detect apoptosis,Transwell was used to detect cell migration and invasion,and Western blot was used to detect protein expressions related to proliferation,apoptosis,invasion,migration,LASP1 and PI3 K/AKT signaling pathway.The eukaryotic expression vector pCDNA-LASP1 and empty vector pCDNA were introduced into SW480 cells by LipofectamineTM2000.The effects of LASP1 overexpression on SW480 proliferation,apoptosis,migration and invasion and PI3 K/AKT signaling pathway were observed.Results:Compared with the blank control group,the SW480 cell viability,cell migration and invasion in the 40μmol/L costunolide group were significantly decreased,and the apoptosis rate was significantly increased,the protein expressions of CDK1,CyclinD1,MMP-2,MMP-9,LASP1,PI3 K,AKT,mTOR were significantly decreased,and the protein expressions of Caspase-3 and Caspase-9 were significantly increased.There were no significant difference between the blank control group and solvent control group.The overexpression of LASP1 reversed the inhibitory effect of costunolide on proliferation,migration and invasion of SW480 cells and the protein expressions of PI3 K,AKT and mTOR,and reversed the apoptosis-promoting effect of costunolide on SW480 cells.Conclusion:Costunolide may regulate the proliferation,apoptosis,invasion and migration of colorectal cancer cells by inhibiting PI3 K/AKT signaling pathway.