不同剂量山奈酚对胰腺癌细胞株PANC-1增殖、凋亡的影响及机制探讨

摘要

Objective To observe the proliferation,apoptosis,and cell cycle distribution of pancreatic cancer PANC-1 cells cultured with different doses of kaempferol, and to explore its mechanism.Methods PANC-1 cells cultured in vitro were divided into the control group and the observation groups 1,2,3,and 4.Cells in the observation groups 1,2, 3,and 4 were added with 20,40,80,and 160 μmol/L kaempferol,while cells in the control group were given the same volume of normal saline.CCK-8 was used to observe the cell proliferation,flow cytometry was used to measure the apopto-sis rate and observe the cell cycle distribution,and Western blotting was applied to detect the intracellular TLR-4,NF-κB, VEGF,and Caspase-3.Results ①With the same time of treatment, the proliferation inhibitory rate of PANC-1 cells gradually increased with the increase of kaempferol concentrations(all P <0.05); with the same concentration of kaempferol,the proliferation inhibitory rate of PANC-1 cells gradually increased with the increase of the treatment time(all P<0.05).②As the kaempferol concentration increased, the proportion of PANC-1 cells in the S phase gradually in-creased(all P<0.05),and the proportion of PANC-1 cells in the G0/G1phase decreased gradually(all P<0.05).③With the increase concentrations of kaempferol,the apoptosis rate of PANC-1 cells gradually increased(all P<0.05).④With the increased concentrations of kaempferol, the relative expression of TLR-4, NF-κB and VEGF in PANC-1 cells gradually decreased(all P <0.05).Conclusions The proliferation of pancreatic cancer PANC-1 cells cultured with kaempferol is inhibited,the apoptosis rate increases,and the proportion of cells in the S phase increases with a dose-de-pendent manner.This may be due to that the kaempferol culture leads to the decreased expression of TLR -4,NF-κB,and VEGF,and the increased expression of Caspase-3 in pancreatic cancer PANC-1 cells.%目的 观察不同剂量山奈酚培养的胰腺癌PANC-1细胞增殖、凋亡和周期分布情况,并探讨其机制.方法 将体外培养的PANC-1细胞分为对照组和观察1、2、3、4组,观察1、2、3、4组分别加入20、40、80、160 μmol/L的山奈酚,对照组加入等量生理盐水.采用CCK-8法观察细胞增殖情况,采用流式细胞术测算细胞凋亡率并观察细胞周期分布,采用Western blotting法检测细胞内TLR-4、NF-κB、VEGF和Caspase-3.结果 ①作用相同时间的情况下,随着山奈酚药物浓度的递增,PANC-1细胞增殖抑制率逐渐升高(P均<0.05);在山奈酚浓度相同的情况下,随着作用时间增加,PANC-1细胞增殖抑制率逐渐升高(P均<0.05).②随着山奈酚浓度的递增,S期PANC-1细胞的比例逐渐上升(P均<0.05),G0/G1期PANC-1细胞的比例逐渐下降(P均<0.05).③随着山奈酚浓度的增加,PANC-1细胞凋亡率逐渐升高(P均<0.05).④随着山奈酚浓度的增加,PANC-1细胞中TLR-4、NF-κB、VEGF蛋白相对表达量逐渐下降(P均<0.05),Caspase-3蛋白相对表达量逐渐上升(P均<0.05).结论 山奈酚培养的胰腺癌PANC-1细胞增殖被抑制,凋亡率上升,S期细胞比例上升,且此效果呈剂量依赖性.这可能是由于山奈酚培养导致胰腺癌PANC-1细胞TLR-4、NF-κB、VEGF表达下降、Caspase-3表达上升.