小白菊内酯对人非小细胞性肺癌H1975细胞凋亡、侵袭与迁移的影响

Effect of parthenolide on apoptosis, invasion and migration of human non-small cell lung cancer H1975 cells

目的探究小白菊内酯(parthenolide, PTL)对非小细胞性肺癌细胞株H1975凋亡、侵袭和迁移的影响及其可能机制。方法 MTT法和集落克隆实验检测PTL对H1975细胞增殖的影响;Annexin V-FITC/PI双染、流式细胞仪检测PTL对H1975细胞凋亡的影响;Transwell实验检测PTL对H1975细胞侵袭和迁移的影响;Western blot检测细胞凋亡、侵袭和迁移相关蛋白,以及PI3K/Akt信号通路相关蛋白的表达。结果 MTT和集落克隆实验结果显示,随着PTL浓度的增加,H1975细胞的增殖明显受到抑制,与对照组相比,差异均具有统计学意义(P<0.05);Annexin V-FITC/PI双染结果显示,PTL能明显诱导H1975细胞发生凋亡(P<0.01);Transwell实验结果显示,PTL能明显抑制H1975细胞侵袭和迁移(P<0.01);Western blot结果显示,PTL明显上调H1975细胞Bax蛋白表达,下调Bcl-2、HIF-1α、MMP-9、Akt、p-Akt(Ser473)蛋白的表达(P<0.05),同时使caspase-3蛋白发生裂解。结论 PTL能明显诱导非小细胞性肺癌H1975细胞发生凋亡,抑制其侵袭和迁移,作用机制可能与PTL抑制PI3K/Akt信号通路有关。

Aim To explore the effects of parthenolide (PTL) on apoptosis,invasion and migration of NSCLC cell line H1975 as well as the possible mechanism. Methods MTT assay and colony formation assay were used to measure cell proliferation.Flow cytometry with Annexin V-FITC/PI double staining were employed to measure cell apoptosis.Transwell assay was applied to measure cell invasion and migration.The expression of apoptosis-related proteins,invasion and migration-associated proteins and PI3K/Akt signaling pathway-related proteins were detected using Western blot. Results MTT assay and colony formation assay results showed that the proliferation of H1975 cells was significantly inhibited with the increase of PTL concentration,and compared with control group,the differences were statistically significant ( P <0.05).Annexin V-FITC/PI double staining result indicated that PTL induced apoptosis in H1975 cells ( P <0.01).The results of transwell assay demonstrated that PTL significantly inhibited the invasion and migration of H1975 cells ( P <0.01).Western blot analysis revealed that PTL increased the expression of Bax and reduced the expression of Bcl-2,HIF-1α,MMP-9,Akt and p-Akt (Ser473) in H1975 cells ( P <0.05),meanwhile,the cleavage of caspase-3 was detected. Conclusions PTL can significantly induce apoptosis and inhibit the invasion and migration of H1975 cells,and the mechanism may be related to the inhibition of PI3K/Akt signaling pathway.