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目的:验证大黄素联合吉西他滨(gemcitabine,GEM)抑制胰腺癌细胞系PANC-1细胞增殖的作用,探讨大黄素促PANC-1细胞凋亡的抗肿瘤作用机制。方法:MTT方法检测大黄素单药以及与吉西他滨联合应用对PANC-1细胞增殖能力的影响。ELISA方法测定细胞上清IL-6水平。RT-PCR方法检测大黄素处理后PANC-1细胞内侵袭相关基因MMP-9的表达情况。Western blot方法分别检测大黄素以及吉西他滨处理PANC-1后凋亡相关蛋白Bax和Bcl-2的表达水平。结果:MTT检测结果显示,加药处理细胞72h后,大黄素组(40μmol/l)PANC-1细胞抑制率为31%,GEM组(20μmol/l)的抑制率为35%,联合用药组的抑制率为49%,与对照组相比差异有统计学意义(P<0.05)。IL-6浓度大黄素组为(22.41±2.27)ng/ml,联合用药组为(15.44±3.91)ng/ml。实验组均显著低于对照组,差异有统计学意义(P<0.05)。大黄素组、联合用药组MMP-9 mRNA表达水平显著低于对照组,GEM组较对照组相比差异无统计学意义(P>0.05)。大黄素组、GEM组、联合用药组Bax表达水平上调,但Bcl-2差异无统计学意义。大黄素组、GEM组、联合用药组的Bax/Bcl-2分别为2.71±0.25,4.73±0.17,5.72±0.36,均显著高于对照组1.14±0.15,结论:体外研究结果显示大黄素能够抑制肿瘤PANC-1细胞增殖,具有促细胞凋亡的作用,与吉西他滨联合应用具有协同效应。
Objective: To investigate the effect of gemcitabine (GEM) on the proliferation of pancreatic cancer cell line PANC-1 and the anti-tumor effect of emodin on the apoptosis of PANC-1 cells. Methods: MTT assay was used to detect the effects of emodin alone and gemcitabine on the proliferation of PANC-1 cells. ELISA method for determination of cell supernatant IL-6 levels. RT-PCR was used to detect the expression of MMP-9 in PANC-1 cells after emodin treatment. Western blot was used to detect the expressions of Bax and Bcl-2, respectively, in emodin and gemcitabine-treated PANC-1 cells. Results: MTT assay showed that the inhibitory rate of PANC-1 cells was 31% in emodin (40μmol / l) group and 35% in GEM group (20μmol / l) The inhibition rate was 49%, compared with the control group, the difference was statistically significant (P <0.05). The concentration of IL-6 in emodin group was (22.41 ± 2.27) ng / ml and that in combination group was (15.44 ± 3.91) ng / ml. The experimental group were significantly lower than the control group, the difference was statistically significant (P <0.05). The expression of MMP-9 mRNA in emodin group and combination group was significantly lower than that in control group, but there was no significant difference between GEM group and control group (P> 0.05). Emodin group, GEM group, combination group Bax expression increased, but Bcl-2 difference was not statistically significant. The levels of Bax / Bcl-2 in emodin, GEM and combination groups were 2.71 ± 0.25, 4.73 ± 0.17 and 5.72 ± 0.36, respectively, which were significantly higher than those in control group (1.14 ± 0.15). Conclusion: In vitro study showed that emodin can inhibit Proliferation of tumor cells PANC-1, with the role of promoting apoptosis, and gemcitabine combination has a synergistic effect.