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目的研究应用潘生丁(DP)阻断平衡型核苷转运载体(hENTs)后,5-氟尿嘧啶(5-FU)对胰腺癌细胞株Panc-1凋亡及细胞周期的影响。方法将Panc-1细胞分为hENTs未阻断组和hENTs阻断组,hENTs阻断组再根据DP浓度分为5μmol/L DP组和10μmol/L DP组。各组细胞分别在含有1.5×106ng/L 5-FU或不含5-FU的培养液中培养24 h后,流式细胞仪检测细胞的凋亡率和细胞周期改变。结果①各组细胞凋亡检测结果:在含有1.5×106ng/L 5-FU培养液中培养24 h后,5μmol/L及10μmol/L DP组的细胞凋亡率明显高于未阻断组(P<0.05),10μmol/L DP组又明显高于5μmol/L DP组(P<0.05);在不含5-FU的培养液中培养24 h后,各组之间细胞凋亡率比较差异无统计学意义(P>0.05)。②各组细胞周期检测结果:在含有1.5×106ng/L 5-FU培养液中培养24 h后,未阻断组细胞进入合成期(S期)的比例减少,停滞在合成前期(G1期),5μmol/L DP组及10μmol/L DP组的细胞进入合成期(S期)的比例较未阻断组进一步减少(P<0.05),且随着DP浓度的增加,细胞进入合成期(S期)的比例减少得更多(P<0.05),5μmol/L DP组和10μmol/L DP组进入合成期(S期)的比例分别是未阻断组的87.09%和74.06%。5-FU对细胞合成后期(G2期)的影响较小,除5μmol/L DP组较未阻断组G2期细胞数量增加有统计学意义(P<0.05)外,其余各组之间差异均无统计学意义(P>0.05);在不含5-FU的培养液中培养24 h后,各组细胞周期中各期无明显改变,各组之间比较差异均无统计学意义(P>0.05)。结论在胰腺癌细胞株Panc-1中,DP阻断细胞膜上hENTs后,能显著增强5-FU对胰腺癌细胞促凋亡作用及抑制胰腺癌细胞分裂增殖的作用,这种增强作用可能与阻断hENTs后细胞内5-FU浓度提高有关,而与DP本身作用无关。
Objective To investigate the effects of 5-fluorouracil (5-FU) on the apoptosis and cell cycle of pancreatic cancer cell line Panc-1 after dipyridamole (DP) blockade of balanced nucleoside transporter (hENTs) Methods Panc-1 cells were divided into hENTs non-blocking group and hENTs blocking group. The hENTs blocking group was divided into 5μmol / L DP group and 10μmol / L DP group according to the DP concentration. Each group of cells were cultured in 1.5 × 106ng / L 5-FU or 5-FU-free medium for 24 h, the apoptosis rate and cell cycle changes were detected by flow cytometry. Results ①Apoptosis assay in each group: The apoptosis rates in 5μmol / L and 10μmol / L DP groups were significantly higher than those in non-block group after cultured in 1.5 × 106ng / L 5-FU medium for 24 hours (P <0.05). The DP of 10μmol / L group was significantly higher than that of 5μmol / L of DP group (P <0.05). After incubated for 24h in 5-FU-free culture medium, No statistical significance (P> 0.05). ② The results of cell cycle test in each group: The percentage of non-blocking cells in the synthesis phase (S phase) decreased after 24 h incubation in 1.5 × 106 ng / L 5-FU medium, and stagnated in the pre-synthesis phase (G1 phase) , Cells in 5μmol / L DP group and 10μmol / L DP group went into the synthesis phase (S phase) than those in the non-blocked group (P <0.05), and with the increase of DP concentration, (P <0.05). The proportion of 5μmol / L DP group and 10μmol / L DP group into the synthesis phase (S phase) were 87.09% and 74.06% respectively in the non-block group. 5-FU had little effect on the cell cycle (G2 phase), except for 5μmol / L DP group (P <0.05), the difference between the other groups was statistically significant No significant difference (P> 0.05). After culturing in 5-FU-free medium for 24 h, there was no significant change in each cell cycle in all groups, with no significant difference between the groups (P> 0.05). Conclusions DP can significantly enhance the pro-apoptotic effect of 5-FU on pancreatic cancer cells and inhibit the proliferation and proliferation of pancreatic cancer cells in the pancreatic cancer cell line Panc-1 after blocking hENTs on the cell membrane, Disruption of hENTs after intracellular 5-FU concentration increased, and has nothing to do with the DP itself.