黄连素联合吉西他滨对胰腺癌PANC-1细胞增殖及凋亡的影响

Effects of berberine combined with gemcitabine on the proliferation and apoptosis of pancreatic cancer PANC-1 cells

目的探究黄连素(BBR)单药及联合吉西他滨(GEM)对胰腺癌PANC-1细胞增殖及凋亡的影响。方法根据BBR浓度的不同,将PANC-1细胞分为对照1组(未处理)、12.5μmol/L BBR组、25.0μmol/L BBR组和50.0μmol/L BBR组,通过CCK-8法检测BBR单药对PANC-1细胞的抑制率,并计算BBR单药的半数抑制浓度(IC_(50))和药物协同指数(q值),筛选取后续实验的最佳药物浓度;根据实验最佳药物浓度将细胞分为对照2组(未处理)、GEM组、BBR组以及联合组,采用克隆形成实验检测各组细胞的克隆形成数,Western blot检测各组细胞中B细胞白血病/淋巴瘤2凋亡调节因子(Bcl-2)和B淋巴细胞瘤-2相关蛋白X(BAX)的表达。结果当25.0μmol/L BBR联合1μmol/LGEM时,其对PANC-1细胞的IC_(50)为1.604μmol/L,q值为1.42,两药协同效果最佳,此浓度作为后续实验的药物浓度;与对照2组相比,BBR、GEM组均能抑制PANC-1细胞的克隆形成,且二者联合应用对PANC-1细胞克隆形成的抑制作用更明显(P<0.05);与对照2组比较,BBR、GEM组PANC-1细胞中Bcl-2蛋白的表达均减少、BAX蛋白的表达增加,且联合组细胞中Bcl-2表达升高及BAX表达降低更明显(P<0.05)。结论BBR与GEM联合应用能发挥协同作用,可增强对胰腺癌细胞抑制作用,并促进其凋亡。

Objective To explore the effect of berberine(BBR)alone and BBR combined gemcitabine(GEM)on the proliferation and apoptosis of pancreatic cancer PANC-1 cells.Methods According to BBR concentration,PANC-1 cells were divided into control 1(untreated),12.5μmol/L,25.0μmol/L,and 50.0μmol/L BBR groups.CCK-8 assay was used to examine the effect of BBR on cell inhibition.The inhibitory rate,the half inhibitory concentration(IC_(50))and drug synergy index(q value)of BBR on PANC-1 cells were calculated,then the optimal BBR concentration for subsequent experiments was selected.Based on optimal concentration of BBR,cells were divided into control 2(untreated),GEM,BBR,and combined groups.Cologenic formation assay was used to measure the numbers of colony in each group,and Western blot was used to detect the expression levels of B-cell lymphoma-2(Bcl-2)and B lymphoma-2 associated protein X(BAX).Results When 25.0μmol/L BBR was combined with 1μmol/LGEM,the IC_(50) for PANC-1 cells was 1.604μmol/L and the q value was 1.42,leading to the best synergistic effect.This combination was used as subsequent experiments.When compared to control 2 group,BBR and GEM groups inhibited the colony formation of PANC-1 cells,and the combined group had an obvious inhibitory effect on colony formation of PANC-1 cells(P<0.05).BBR and GEM groups had reduced Bcl-2 and increased BAX in PANC-1 cells relative to control 2 group,the combination of BBR with GEM augmented the effect of BBR or GEM(P<0.05).Conclusion The combination of BBR with GEM can exert a synergistic effect,which enhances the inhibitory effect on pancreatic cancer cells and promotes cellular apoptosis.