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目的研究乳腺癌细胞株MDA-MB-231、MDA-MB-468、SK-BR-3、MCF-7中人平衡型核苷载体(hENTs)的表达及其对5-氟尿嘧啶(5-FU)细胞毒性的影响。方法 MDA-MB-231、MDA-MB-468、SK-BR-3、MCF-7细胞常规培养于含10%小牛血清的高糖DMEM培养液中。逆转录-聚合酶链反应(RT-PCR)检测4种细胞株中hENT1 mRNA及hENT2 mRNA的表达。每种细胞分别在含有一定浓度序列(1.28×104 ng/L~2.00×108 ng/L)5-FU的培养液中培养48 h。MTT法检测4种细胞株增殖,并计算其5-FU的半数抑制浓度(IC50)。结果①hENT1及hENT2 mRNA在MDA-MB-231、MDA-MB-468、SK-BR-3细胞中的表达均较其在MCF-7细胞中高(P<0.05),但其表达在MDA-MB-231、MDA-MB-468、SK-BR-3细胞中差异无统计学意义(P>0.05)。hENT2 mRNA表达在4种细胞中均较hENT1 mRNA表达低,差异有统计学意义(P<0.05)。②MTT结果显示,5-FU的IC50在MDAMB-231、MDA-MB-468、SK-BR-3细胞株中均较在MCF-7细胞株中低(P<0.05),在MDA-MB-231、MDA-MB-468、SK-BR-3细胞株之间差异无统计学意义(P>0.05)。③在5-FU的IC50较低的乳腺癌细胞株(MDA-MB-231、MDAMB-468、SK-BR-3)中高表达hENTs,而在5-FU的IC50较高的乳腺癌细胞株(MCF-7)中,低表达hENTs或无表达。结论在乳腺癌细胞株中,细胞膜上hENTs的表达情况能显著影响5-FU的作用效果,其结果提示,在临床上可能需要依据hENTs的表达情况来选择核苷类抗癌药物。
Objective To study the expression of hENTs in breast cancer cell lines MDA-MB-231, MDA-MB-468, SK-BR-3 and MCF-7 and its effect on 5-fluorouracil (5-FU) Cytotoxic effects. Methods MDA-MB-231, MDA-MB-468, SK-BR-3 and MCF-7 cells were cultured in high glucose DMEM containing 10% calf serum. The expression of hENT1 mRNA and hENT2 mRNA in 4 cell lines were detected by reverse transcription-polymerase chain reaction (RT-PCR). Each cell line was cultured for 48 h in a medium containing a concentration of 5-FU (1.28 × 104 ng / L ~ 2.00 × 108 ng / L). The proliferation of 4 cell lines was detected by MTT assay and the half inhibitory concentration (IC50) of 5-FU was calculated. Results ① The expressions of hENT1 and hENT2 mRNA in MDA-MB-231, MDA-MB-468 and SK-BR-3 cells were higher than those in MCF-7 cells 231, MDA-MB-468 and SK-BR-3 cells. There was no significant difference between the two groups (P> 0.05). The expression of hENT2 mRNA was lower than that of hENT1 mRNA in all four cell types (P <0.05). ② MTT results showed that the IC50 of 5-FU was lower (P <0.05) in MDAMB-231, MDA-MB-468 and SK-BR-3 cell lines than in MCF- , MDA-MB-468 and SK-BR-3 cell lines without statistical significance (P> 0.05). ③ High expression of hENTs in 5-FU cells with low IC50 (MDA-MB-231, MDAMB-468, SK-BR-3) MCF-7), low expression of hENTs or no expression. Conclusion The expression of hENTs on the cell membrane in breast cancer cell lines can significantly affect the effect of 5-FU. The results suggest that nucleoside anticancer drugs may need to be selected according to the expression of hENTs in clinic.