重楼皂苷Ⅶ对胰腺癌PANC-1细胞增殖、迁移与侵袭作用及机制研究

Effect and mechanism of polyphyllin Ⅶ on proliferation, migration and invasion of pancreatic cancer PANC-1 cells

目的研究重楼皂苷Ⅶ对人胰腺癌PANC-1细胞增殖、迁移与侵袭作用及机制。方法采用MTT法检测重楼皂苷Ⅶ对PANC-1细胞存活率的影响;观察重楼皂苷Ⅶ对PANC-1细胞形态的影响;采用划痕实验和Transwell小室法考察重楼皂苷Ⅶ对PANC-1细胞迁移和侵袭能力的影响;采用流式细胞术检测重楼皂苷Ⅶ对PANC-1细胞凋亡的影响;采用Western blotting法检测重楼皂苷Ⅶ对PANC-1细胞半胱氨酸蛋白酶-3(Caspase-3)、B细胞淋巴瘤-2(B-celllymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-associated X protein,Bax)、聚腺苷二磷酸核糖聚合酶(poly ADP-ribose polymerase,PARP)、DNA酶抑制物(inhibitor of caspase-activated deoxyribonuclease,ICAD)和程序性死亡分子配体-1(programmed death ligand 1,PD-L1)蛋白表达的影响。结果重楼皂苷Ⅶ可抑制PANC-1细胞存活率,显著抑制PANC-1细胞迁移和侵袭能力(P<0.01),显著诱导PANC-1细胞凋亡(P<0.05、0.01),显著升高PANC-1细胞中凋亡相关蛋白Cleaved Caspase-3、PARP、Bax表达水平(P<0.01),显著降低pro-PARP、ICAD、Bcl-2、PD-L1蛋白表达水平(P<0.05、0.01)。结论重楼皂苷Ⅶ能够抑制PANC-1细胞增殖、迁移和侵袭,并可能通过下调PD-L1表达诱导PANC-1细胞凋亡。

Objective To study the effect and mechanism of polyphyllin Ⅶ on proliferation, migration and invasion of human pancreatic cancer PANC-1 cells. Methods MTT assay was used to detect the effect of polyphyllin Ⅶ on viability of PANC-1 cells. Effect of polyphyllin Ⅶ on morphology of PANC-1 cells was observed. Effect of polyphyllin Ⅶ on cell migration and invasion were detected by wound scratch assay and Transwell chamber. Effect of polyphyllin Ⅶ on apoptosis was detected by flow cytometry. Western blotting was used to detect the effect of polyphyllin Ⅶ on expressions of cysteine protease-3(Caspase-3), B-cell lymphoma-2(Bcl-2), Bcl-associated X protein(Bax), poly ADP-ribose polymerase(PARP), inhibitor of caspase-activated deoxyribonuclease(ICAD), and programmed death ligand 1(PD-L1) in PANC-1 cells. Results Survival rate of PANC-1 cells was inhibited, migration and invasion of PANC-1 cells were significantly inhibited(P < 0.01), cell apoptosis was significantly induced(P < 0.05, 0.01), expressions of apoptosis-related proteins Cleaved Caspase-3, PARP, Bax were significantly increased(P < 0.01), and expressions of pro-PARP, ICAD, Bcl-2, and PD-L1 were significantly reduced by polyphyllin Ⅶ(P < 0.05, 0.01). Conclusion Polyphyllin Ⅶ can inhibit proliferation, migration and invasion of PANC-1 cells, and may induce PANC-1 cell apoptosis by down-regulating PD-L1 expression.