葡萄籽原花青素联合吉西他滨对胰腺癌细胞增殖与凋亡的研究

Effect of grape seed proanthocyanidins extract combined with gemcitabine on the proliferation and apoptosis of pancreatic cancer cell

目的观察葡萄籽原花青素(GSPE)联合吉西他滨对胰腺癌PANC-1细胞增殖与凋亡的影响及机制。方法将PANC-1细胞分为对照组-1(20μg·m L-1吉西他滨)、对照组-2(75μg·m L-1GSPE)、实验组(20μg·m L-1吉西他滨+75μg·m L-1GSPE)和模型组(无任何处理)。药物干预24 h后,用溴化四唑蓝(MTT)法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,用反转录聚合酶链式反应(RT-PCR)检测microRNA-27a(miR-27a)相对表达水平。结果干预24 h后,模型组、对照组-1、对照组-2和实验组细胞存活率分别为(98.67±0.12)%,(48.67±9.45)%,(43.52±6.44)%,(13.05±4.25)%,与模型组比较,对照组-1、对照组-2及实验组细胞存活率均降低(均P<0.05),且实验组细胞存活率低于阳性对照组(P<0.05)。干预48 h后,对照组-1、对照组-2及实验组细胞增殖指数均低于模型组,凋亡率高于模型组(均P<0.05);且实验组增殖指数低于对照组-1、对照组-2,细胞凋亡率高于对照组-1、对照组-2(均P<0.05)。干预48 h后,模型组、对照组-1、对照组-2及实验组miR-27a相对表达量分别为1.00±0.00,0.69±0.19,0.56±0.14,0.38±0.15,对照组-1、对照组-2及实验组miR-27a相对表达量均低于模型组(均P<0.05),实验组miR-27a相对表达量低于对照组(P<0.05)。结论 GSPE联合吉西他滨可抑制胰腺癌PANC-1细胞增殖,并促其凋亡,其机制可能与下调miR-27a表达有关。

Objective To observe the effect of grape seed proanthocyanidins extract（ GSPE） combined with gemcitabine on the proliferation and apoptosis of pancreatic cancer PANC-1 cell and its mechanism.Methods PANC-1 cells were divided into control group-1（ 20μg·m L^（-1） gemcitabine）,control group-2（ 75 μg·m L^（-1） GSPE）,test group（ 20 μg · m L^（-1） gemcitabine ＋ 75 μg · m L^（-1） GSPE） and model group（ without any treatment）. The cell proliferation was detected by methylthiazolyldiphenyl-tetrazolium（ MTT） assay after 24 h of drug intervention,flow cytometry was used to detect the cell apoptosis,the relative expression level of microRNA-27 a（ miR-27 a） was detected by reserve transcription-polymerase chain reaction（ RT-PCR）. Results After 24 h intervention,the cell viabilities in model group,control group-1,control group-2 and test group were（ 98. 67 ± 0. 12） %,（ 48. 67 ± 9. 45） %,（ 43. 52 ± 6. 44） %,（ 13. 05 ± 4. 25） %. Compared with model group,the cell viability of control group-1,control group-2 and test group decreased（ all P 0. 05）,the cell viability of test group was lower than those in control group-1 and control group-2（ P 0. 05）. After 48 h intervention,cell proliferation indexes of control group-1 and control group-2 and test group were lower than that in model group,the apoptosis rates were higher than that of model group（ all P 0. 05）; and proliferation index in test group was lower than those in control group-1 and control group-2,the apoptosis rate was significantly higher than those in control group-1 and control group-2（ all P 0. 05）. After 48 h of intervention,the relative expressions of miR-27 a in model group,control group-1,control group-2 and test group were 1. 00 ± 0. 00,0. 69 ± 0. 19,0. 56 ± 0. 14,0. 38 ± 0. 15,the relative expression levels of miR-27 a in control group-1 and control group-2 and test group were lower than that in model group,the relative expression of miR-27 a in test group was lower than those in control group-1 and control group-2（ all P 0. 05）.Conclusion GSPE combined with gemcitabine can inhibit pancreatic cancer PANC-1 cell proliferation and promote apoptosis,the mechanism may be related to down-regulation of miR-27 a expression.