苦参碱对胰腺癌CFPAC-1细胞增殖、凋亡及Hedgehog信号通路的影响

Effect of matrine on Hedgehog signaling pathway, cell proliferation and apoptosis in pancreatic carcinoma CFPAC-1 cells

目的:探讨苦参碱对人胰腺癌CFPAC-1细胞增殖、凋亡的影响及其可能的分子机制.方法:应用MTT法检测苦参碱对人胰腺癌CFPAC-1细胞增殖的抑制作用,流式细胞仪分析细胞的凋亡率,Western blot法检测苦参碱对Hedgehog(Hh)通路中Ptch、Smo、Gli-l、Bcl-2蛋白表达的影响.结果:MTT实验显示:经浓度为0.25、0.50、1.00、2.00、4.00 mg/mL的苦参碱作用6、12、24、48 h后,苦参碱对CFPAC-1细胞的生长抑制作用呈明显的浓度依赖性和时间依赖性(P<0.05),其中24 h的IC50为1.266 mg/mL.FCM分析显示与对照组(8.2367%±1.685%)比较,不同浓度苦参碱作用24 h后CFPAC-1细胞的凋亡率分别11.3867%±0.4652%、13.6167%±0.5785%、17.0433%±0.72501%、23.8700%±3.31107%、46.1333%±1.61398%,均高于对照组(P<0.05).其中除药物浓度为0.25 mg/mL与0.50 mg/mL浓度组比较时,无统计学意义(P>0.05),余浓度组两两对比均有统计学意义(P<0.05).Western blot结果显示:1.00 mg/mL苦参碱作用24 h后,CFPAC-1细胞的Bcl-2、Gli-l、Ptch、Smo蛋白表达均显著低于阴性对照组(F值分别为:15.557,8.745,24.386,12.768,P<0.05).结论:苦参碱能够有效地抑制胰腺癌细胞的增殖并促进其凋亡,其机制可能与Hh通路及其下游的Bcl-2有关.

AIM： To assess the effect of matrine on the proliferation and apoptosis of human pancreatic carcinoma CFPAC-1 cells and the potential molecular mechanism.METHODS： MTT assay was used to detect the inhibitory effect of matrine on the proliferative rate of CFPAC-1 cells. Flow cytometry was performed to evaluate cell apoptosis. Western blot was applied to determine the expression levels of Hedgehog signaling pathway molecules such as Ptch, Smo, Gli-l and Bcl-1.RESULTS： Matrine treatment dose- and time-dependently suppressed proliferation of CFPAC-1 cells（P〈0.05）. IC50 of matrine at 24 h was 1.266 mg/mL. Compared with control cells, treatment with matrine at a concentration of 0.25, 0.50, 1.00, 2.00, or 4.00 mg/mL for 24 h significantly increased cell apoptosis, and the apoptosis rates were 8.2367% ± 1.685%,11.3867% ± 0.4652%, 13.6167% ± 0.5785%, 17.0433% ± 0.72501%, 23.8700% ± 3.31107% and 46.1333% ± 1.61398%, respectively（P〈0.05）. There was a significant difference in cell apoptosis between matrine treated groups（P〈0.05） except for the comparison between the concentrations of 0.25 mg/mL and 0.50 mg/mL. Western blot analysis indicated that compared with control cells, protein expression of Bcl-2, Gli-l, Ptch and Smo was significantly decreased in CFPAC-1 cells treated with 1 mg/mL matrine for 24 h（F = 15.557, 8.745, 24.386, 12.768, P〈0.05）.CONCLUSION： Matrine can effectively inhibit proliferation and induce apoptosis of human pancreatic carcinoma CFPAC-1 cells, via mechanisms possibly involving the Hedgehog signal pathway and Bcl-2.