低糖高乳酸环境下吉非替尼诱导HeLa细胞EGFR-TKI耐药的研究

发布时间：2019-06-11 06:00

【摘要】：目的探讨低糖高乳酸微环境对表皮生长因子酪氨酸激酶抑制剂(EGFR-TKI)抑制He La细胞的影响和可能机制。方法 He La细胞在常糖(葡萄糖10 mmol/L)和低糖高乳酸(葡萄糖3 mmol/L+乳酸2.5 mmol/L)环境下培养,并分别给予2.67μmol/L吉非替尼干预。采用CCK-8法测定He La细胞生长抑制率,流式细胞术检测细胞周期,荧光定量RT-PCR检测EGFR和m TOR m RNA水平的表达。结果与常糖组比较,低糖乳酸组24 h和72 h细胞抑制率均显著升高(P0.01),48 h细胞抑制率略高于常糖组。与吉非替尼阴性组比较,常糖+吉非替尼组和低糖乳酸+吉非替尼组在24、48、72 h三个时点细胞抑制率均显著上升(P0.01)。与常糖组相比,低糖乳酸组48 h细胞诱导凋亡率显著上升(P0.01),低糖乳酸+吉非替尼组细胞诱导凋亡率较低糖乳酸组显著降低(P0.01)。与常糖组比较,低糖乳酸组存活细胞的EGFR和m TOR m RNA表达水平上调(P0.05)。常糖+吉非替尼组的EGFR和m TOR m RNA水平均上调(P0.05)。与低糖乳酸组比较,低糖乳酸+吉非替尼组的EGFR和m TOR m RNA上调水平有显著差异(P0.01)。结论高乳酸低糖环境下吉非替尼可大幅度上调存活He La细胞EGFR和m TOR表达水平,可能是诱导He La细胞抵抗EGFR-TKI的机制。
[Abstract]:Objective to investigate the effect and possible mechanism of low glucose and high lactic acid microenvironment on the inhibition of He La cells by epidermal growth factor tyrosine kinase inhibitor (EGFR-TKI). Methods He La cells were cultured in normal glucose (glucose 10 mmol/L) and low glucose high lactic acid (glucose 3 mmol/L lactic acid 2.5 mmol/L) environment and treated with 2.67 渭 mol / L gefitinib, respectively. The growth inhibition rate of He La cells was measured by CCK- 8 assay, the cell cycle was detected by flow cytometry, and the expression of EGFR and m TOR m RNA was detected by fluorescence quantitative RT-PCR. Results compared with normal glucose group, the inhibition rate of cells in low glucose lactic acid group increased significantly at 24 h and 72 h (P 0.01), and the cell inhibition rate at 48 h was slightly higher than that in normal glucose group. Compared with gefitinib negative group, the cell inhibition rate of normal glucose gifetene group and low glucose lactic acid gifitinib group increased significantly at 24 h, 48 h and 72 h (P 0.01), and the cell inhibition rate was significantly increased at 24 h, 48 h and 72 h, respectively (P 0.01). Compared with normal glucose group, the apoptosis rate of cells in low glucose lactic acid group increased significantly at 48 h (P 0.01), and the apoptosis rate in low glucose lactic acid group was significantly lower than that in low glucose lactic acid group (P 0.01). Compared with normal glucose group, the expression of EGFR and m TOR m RNA in survival cells of low glucose lactic acid group was up-regulated (P 0.05). The levels of EGFR and m TOR m RNA in giferinib group were up-regulated (P 0.05). Compared with the low glucose lactic acid group, the up-regulation levels of EGFR and m TOR m RNA in the low glucose lactic acid group were significantly different (P 0.01). Conclusion gefitinib can significantly up-regulate the expression of EGFR and m-TOR in survival He La cells in high lactic acid and low glucose environment, which may be the mechanism of inducing He La cells to resist EGFR-TKI.
【作者单位】：福建医科大学附属泉州市第一医院;北京中医药大学;厦门大学;
【基金】：泉州市第一医院青年科研(2014-QN-24) 泉州市科技计划项目(2015241) 国家面上项目(81672094)
【分类号】：R737.33

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