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目的探讨EGFR和HER-3基因在人肺腺癌细胞株A549和培美曲塞二钠(PEM)诱导的耐药细胞株A549/PEM中的表达变化。方法用大剂量PEM诱导、建立耐药细胞株A549/PEM,CCK-8法检测A549和A549/PEM细胞的增殖抑制率,流式细胞术检测细胞的周期分布,RT-PCR检测EGFR与HER-3 mRNA在A549和A549/PEM细胞中的表达量。结果 CCK-8法测得PEM对A549细胞的半数抑制浓度(IC50)为3.43×10-6mol/L,用PEM反复处理细胞4个月,成功诱导出耐药指数(RI)为5.15倍的耐药细胞株A549/PEM。流式细胞术检测结果显示,A549和A549/PEM细胞处于G1/G0期的比例分别为(56.25±0.10)%和(65.12±0.40)%(P<0.05),处于S期的细胞比例分别为(40.87±0.30)%和(31.74±0.40)%(P<0.05)。EGFR mRNA在A549和A549/PEM细胞中的相对表达量分别为0.55±0.08和0.64±0.07,HER-3 mRNA在A549和A549/PEM细胞中的相对表达量分别为1.29±0.03和1.56±0.12,其差异均有统计学意义(P<0.05)。结论与A549细胞相比,A549/PEM中EGFR和HER-3的表达量均增高,提示这两种基因表达量的变化可能与PEM耐药相关。
Objective To investigate the expression changes of EGFR and HER-3 genes in human lung adenocarcinoma cell line A549 and pemetrexed disodium (PEM) -mediated drug-resistant cell line A549 / PEM. Methods The cell line A549 / PEM was induced by large dose of PEM. The proliferation inhibition rate of A549 and A549 / PEM cells was detected by CCK-8 assay. The cell cycle distribution was detected by flow cytometry. The expressions of EGFR and HER- 3 mRNA expression in A549 and A549 / PEM cells. Results The IC50 of PEM to A549 cells was 3.43 × 10-6 mol / L as determined by CCK-8. The cells were repeatedly treated with PEM for 4 months and the resistance index (RI) was 5.15 times Drug cell line A549 / PEM. Flow cytometry showed that the proportion of A549 and A549 / PEM cells in G1 / G0 phase was (56.25 ± 0.10)% and (65.12 ± 0.40)%, respectively (P <0.05). The percentage of cells in S phase were (40.87 ± 0.30)% and (31.74 ± 0.40)%, respectively (P <0.05). The relative expression levels of EGFR mRNA in A549 and A549 / PEM cells were 0.55 ± 0.08 and 0.64 ± 0.07, respectively. The relative expression levels of HER-3 mRNA in A549 and A549 / PEM cells were 1.29 ± 0.03 and 1.56 ± 0.12, The differences were statistically significant (P <0.05). Conclusions Compared with A549 cells, the expression of EGFR and HER-3 in A549 / PEM are both increased, suggesting that the changes of the expression of these two genes may be related to the resistance of PEM.