防己诺林碱诱导人乳腺癌细胞MDA-MB-231凋亡的作用机制

目的 探讨防己诺林碱(FAN)对三阴性乳腺癌(TNBC)的抗肿瘤机制.方法 体外细胞培养人乳腺癌细胞MDA-MB-231,Alamar-Blue法检测FAN对人乳腺癌细胞MDA-MB-231的半抑制浓度(IC50);6孔板检测细胞迁移情况;细胞流式技术检测细胞凋亡情况;Western Blot检测磷脂酰肌醇-3羟基激酶(PI3K)、蛋白激酶B(AKT)、哺乳类动物雷帕霉素靶蛋白(mTOR)及磷酸化PI3K、AKT、mTOR蛋白表达.结果 FAN可抑制人乳腺癌细胞MDA-MB-231的活力(IC50为6.25μmol/L),抑制人乳腺癌细胞MDA-MB-231的迁移能力,且随着FAN浓度升高,抑制作用明显.FAN可以诱导人乳腺癌细胞MDA-MB-231凋亡,且随着FAN浓度升高,细胞凋亡率增高,同时FAN还可以下调PI3K、AKT、mTOR及磷酸化PI3K、AKT、mTOR蛋白的表达,随药物浓度的升高,其蛋白表达降低.结论 FAN可通过下调TNBC MDA-MB-231细胞凋亡PI3K/AKT/mTOR信号通路,抑制TNBC细胞的增殖、迁移,诱导细胞凋亡,可能具有抗肿瘤作用.

The mechanism of fangchinoline in inducing the apoptosis of MDA-MB-231 human breast cancer cells

Objective This study was conducted to explore the anti-tumor mechanism of fangchinoline (FAN) against triple negative breast cancer (TNBC). Method MDA-MB-231 human breast cancer cells were incubated in vitro. The half maximal inhibitory concentration (IC50) of FAN against MDA-MB-231 human breast cancer cells was detected by Alamar-Blue assay. Cell migration was detected by wound scratch assay and apoptotic cells were detected by Flow cy-tometry;the expression of phosphatidylinositol 3-hydroxy kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR) and phosphorylated PI3K, AKT, mTOR protein were detected by western blot. Result FAN inhibit-ed the activity and migration of MDA-MB-231 human breast cancer cells (IC50 is 6.25μmol/L), and the inhibitory effect was obvious with the increase of FAN concentration;FAN induced cell apoptosis of the MDA-MB-231 human breast can-cer cells,and the apoptosis rate increased with the increase of FAN concentration;meanwhile, FAN downregulated the ex-pression of PI3K/AKT/mTOR protein and phosphorylated PI3K/AKT/mTOR protein, and these protein concentrations were reduced with the increase of FAN concentration. Conclusion FAN achieves potential anti-tumor effects by down-regulating TNBC MDA-MB-231 cell apoptosis PI3K/AKT/mTOR signaling pathways and inhibiting the proliferation and migration of TNBC cells, as well as inducing cell apoptosis.