血糖对EGFR突变肺腺癌患者靶向治疗耐药的影响及其机制

doi:10.3969/j.issn.1004-616x.2024.05.005

摘要/Abstract

摘要： 目的： 探讨血糖因素对表皮生长因子受体(EGFR)突变肺腺癌患者靶向治疗耐药的影响及其机制。方法： 以2018—2019年于首都医科大学附属北京胸科医院接受靶向治疗的277例肺腺癌患者为研究对象，分析患者靶向治疗耐药时的空腹血糖值(FBG)及其与进展时间的关系。另外，利用高糖和改良普通培养基模拟体外血糖环境对肺腺癌细胞HCC827及其衍生的耐药细胞株AZDR (对EGFR抑制剂AED9291不敏感)进行培养，观察两种细胞的生长情况。再根据细胞培养环境的葡萄糖含量以及是否添加AZD7291药物，设置高糖组、普通组、高糖+AZD处理组和普通+AZD处理组共4组，分别利用细胞集落染色和CCK-8技术检测两种细胞的增殖情况；采用流式细胞术检测细胞凋亡情况；使用Western blot检测细胞凋亡因子caspase-3和代谢相关蛋白mTOR的表达变化；采用RNA测序技术分析AZDR细胞获得耐药后的胰岛素受体及葡萄糖代谢相关基因表达变化。结果： 临床样本结果显示，在EGFR-TKI治疗发生耐药时，除了糖尿病合并肺癌患者(37人) FBG升高之外，单纯肺腺癌患者(240人)的FBG也升高，17.4%的肺腺癌患者超过健康人FBG标准范围的上限(6.1 mmol/L)；对肺腺癌患者的生存分析发现，FBG值大于6.1 mmol/L的患者比FBG值在4.0~6.0 mmol/L的患者更晚发生耐药(P<0.05)。体外细胞培养发现，耐药细胞株AZDR在高糖培养环境下生长缓慢，而亲本细胞HCC827生长不受高糖影响。相比亲本细胞HCC827，耐药细胞AZDR在高糖环境下凋亡发生率升高(P<0.05)，凋亡相关蛋白caspase-3表达升高(P<0.05)，胰岛素受体IGFBP7、IGFBP2、IGF1R和IGF2R表达升高；葡萄糖代谢相关基因SLC16A2、SLC2A13和HK2降低(P<0.05)，而mTOR和HK1基因表达水平升高(P<0.05)。结论： 肺腺癌患者血糖水平变化可能是影响患者预后和耐药性进展的重要因素。高血糖环境不利于耐药肿瘤细胞亚群的生存，可能缘于其胰岛素受体、葡萄糖代谢基因的表达变化。在肺腺癌靶向治疗过程中需要辩证地看待患者的血糖变化。

关键词: 肺腺癌, 空腹血糖, 靶向治疗, 酪氨酸激酶抑制剂, 肿瘤耐药

Abstract: OBJECTIVE： To investigate the control of blood glucose factors on efficacy of targeted therapy in patients with epidermal growth factor receptor(EGFR)- mutated lung adenocarcinoma. METHODS： From 2018 to 2019，a total of 277 lung adenocarcinoma patients who received the targeted therapy at the Beijing Chest Hospital，Capital Medical University，were recruited for our study. The relationships among fasting blood glucose (FBG) levels，resistance to targeted therapy and progression-free survival were analyzed. In addition，a culture medium with different glucose concentrations was used to simulate the in vitro glucose environment，and the effects of glucose on the growth of lung adenocarcinoma cell line HCC827 and its derived resistant cell line AZDR were investigated using cell biology methods. According to the glucose content of the cell culture environment，and whether AZD drugs were added or not，four in vitro groups were set up：high glucose，ordinary，high glucose +AZD treatment，and ordinary +AZD treatment groups. Cell proliferation was assessed using colony staining and CCK-8 assays. Cell apoptosis was detected using flow cytometry. Changes in expression of apoptosis-related factors (caspase-3 and metabolism-related protein mTOR) were evaluated using Western blot. RNA sequencing was performed to analyze the gene expression changes in AZDR cells after acquiring resistance. RESULTS： Among patients who showed EGFR-TKI treatment resistance，in addition to the increase of FBG in patients with diabetes combined with lung cancer (37 patients)，FBG in patients with simple lung adenocarcinoma (240 patients) also increased，and 17.4% of lung adenocarcinoma patients exceeded the upper limit of the standard range of FBG in healthy people (6.1 mmol/L). Survival analysis revealed that patients with FBG levels greater than 6.1 mmol/L experienced resistance to targeted therapy later，compared to patients with FBG levels in the range of 4.0-6.0 mmol/L (P<0.05). The cell culture study revealed that the resistant cell line AZDR exhibited slow growth in a high-glucose environment，while the parental cell line HCC827 was not affected. Compared to the parental cell line HCC827，the resistant cell line AZDR showed increased apoptosis rate (P<0.05)，upregulation of apoptosis-related protein caspase-3 (P<0.05)，increased expression of insulin receptor IGFBP7，IGFBP2，IGF1R，and IGF2R，decreased expression of glucose metabolism-related genes SLC16A2，SLC2A13，and HK2，and upregulation of mTOR and HK1 in a high-glucose environment (P<0.05). CONCLUSION： Changes of blood glucose level in patients with lung adenocarcinoma may be an important factor affecting the prognosis and the progression of drug resistance. In vitro studies showed that hyperglycemic environment is not conducive to the survival of drug resistant tumor cell subpopulations，which may be caused by changes in the expression of insulin receptors and glucose metabolism genes. In the process of targeted therapy for lung adenocarcinoma，it may be necessary to dialectically control changes of blood glucose in patients.

Key words: lung adenocarcinoma, fasting blood glucose, targeted therapy, tyrosine kinase inhibitors, tumor resistance

中图分类号:

R734.2

张鑫桐, 顾勐, 王子宇, 韦攀健, 王敬慧, 谭金晶. 血糖对EGFR突变肺腺癌患者靶向治疗耐药的影响及其机制[J]. 癌变·畸变·突变, 2024, 36(5): 365-372,383.

ZHANG Xintong, GU Meng, WANG Ziyu, WEI Panjian, WANG Jinghui, TAN Jinjing. Control of blood glucose on targeted therapy in EGFR-mutated lung adenocarcinoma[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(5): 365-372,383.

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