白藜芦醇对人胰腺癌PANC-1细胞增殖与侵袭的影响

目的 探讨白藜芦醇对人胰腺癌PANC-1细胞增殖与侵袭能力的影响.方法 实验分为空白对照组,0.1%DMSO组,白藜芦醇组(50、100、200 μmol/L).MTT法检测白藜芦醇对细胞增殖的影响;流式细胞仪检测细胞凋亡及周期变化;Transwell侵袭小室检测白藜芦醇对细胞侵袭的影响;荧光实时定量PCR和Western blot检测白藜芦醇对细胞Bax、Bcl-2、MMP-2、MMP-9表达的影响.数据以-x±s表示,多组比较采用方差分析.结果 (1)空白对照组抑制率为0;0.1%DMSO组细胞抑制率为3.25%±0.42%;白藜芦醇50 μmol/L组细胞抑制率为13.23%±1.68%;白藜芦醇100μmol/L组细胞抑制率为42.25%±3.20%;白藜芦醇200μmol/L组细胞抑制率为56.94%±5.31%.各组比较,差异有统计学意义(F=460.10,P<0.05).(2)空白对照组细胞凋亡率为0.05%±0.03%;0.1%DMSO组细胞为凋亡率为3.39%±1.77%;白藜芦醇50 μmol/L组细胞凋亡率为6.92%±1.85%;白藜芦醇100 μmol/L组细胞凋亡率为19.05%±2.01%;白藜芦醇200 μmol/L组细胞凋亡率为27.17%±6.43%.各组比较,差异有统计学意义(F=38.84,P<0.05).(3)0.1%DMSO组对细胞周期无显著影响.白藜芦醇引起PANC-1细胞G0/G1期和S期阻滞,G2/M期细胞减少.(4)空白对照组平均穿膜细胞数为61±13;0.1%DMSO组为54±13;白藜芦醇50 μmol/L组为48±15;白藜芦醇100 μmol/L组为23±6;白藜芦醇200 μmol/L组为18±7.各组比较,差异有统计学意义(F=69.08,P<0.05).(5)白藜芦醇可使PANC-1细胞Bax表达升高,Bcl-2表达下调.MMP-2、MMP-9的表达明显受到抑制,mRNA和蛋白水平变化一致.结论 白藜芦醇可明显抑制胰腺癌PANC-1细胞增殖,诱导细胞凋亡并抑制其侵袭能力.

Effect of resveratrol on the proliferation and invasion of human pancreatic cancer PANC-1 cells

Objective To investigate the effect of resveratrol on the proliferation and invasion of human pancreatic cancer PANC-1 cells. Methods Five groups including blank control group, 0. 1% dimethylsulfoxide (DMSO) group and resveratrol groups (50, 100, 200 μmol/L) were established. The proliferation of PANC-1 cells was detected by MTT assay. The apoptosis and cell cycle change were analyzed by flow cytometry. The invasive ability of PANC-1 cells was observed with a Transwell cell culture chamber. The expressions of Bax, Bcl-2,matrix metalloproteinases 2 (MMP-2) and 9 (MMP-9) of the PANC-1 cells were assayed by real-time quantitative PCR and Western blot. All data were analyzed using the analysis of variance. Results ( 1 ) The inhibition rate of resveratrol on the proliferation of PANC-1 cells was 0 in the blank control group, 3.25% ±0.42% in the 0. 1% DMSO group, 13.23% ± 1.68% in the 50 μmol/L of resveratrol group, 42.25% ± 3.20% in the 100 μmol/L of resveratrol group, and 56.94% ±5.31% in the 200 μmol/L of resveratrol group. There was a significant difference in the inhibition rate among the five groups (F=460. 10, P<0.05). (2) The apoptosis rate was 0.05% ±0.03% in the blank control group, 3.39% ± 1.77% in the 0. 1% DMSO group, 6.92% ± 1.85% in the 50 μmol/L of resveratrol group, 19.05% ± 2.01% in the 100 μmol/L of resveratrol group, and 27. 17% ±6.43% in the 200 μmol/L of resveratrol group. There was a significant difference in the apoptosis rate among the five groups (F = 38.84, P < 0.05). (3) There was no significant effect of 0. 1% DMSO on the cell cycle of PANC-1 cells. The number of PANC-1 cells in the G0/G1 and S phase was increased. (4) The average number of invading PANC-1 cells was 61 ± 13 in the blank control group, 54 ± 13 in the 0. 1% DMSO group, 48 ± 15 in the 50 μmol/L of resveratrol group, 23 ±6 in the 100 μ mol/L of resveratrol group and 18 ±7 in the 200 μmol/L of resveratrol group. There was a significant difference in the number of invading PANC-1 cells among the five groups (F = 69.08, P < 0.05 ). (5) There were up-regulated mRNA and protein expressions of Bax and down-regulated mRNA and protein expressions of Bcl-2, and the expressions of MMP-2 and MMP-9 of the PANC-1 cells were inhibited in the resveratrol groups. The changes of the protein expressions of Bax, Bcl-2, MMP-2, MMP-9 were consistent with the changes of the mRNA expressions of the four indexes. Conclusion Resveratrol can significantly inhibit the proliferation and invasion, as well as induce apoptosis of PANC-1 cells in vitro.