Abstract

Context

Yang-Yin-Jie-Du Decoction (YYJDD) was used to improve gefitinib efficacy in our clinical practice, but its mechanism remains unclear.

Objective

This study explored if YYJDD could reverse gefitinib resistance.

Materials and methods

H1975 cells were exposed to control, 10 μM gefitinib, 3.2 mg/mL YYJDD or combination treatment. Cell viability was detected by MTT during 0–96 h. Apoptosis and the PI3K/Akt proteins were tested by flow cytometry and western-blot at 24 h. LY294002 was applied to further determine the role of the PI3K/Akt. 23 BALB/c nude xenograft mice received normal saline (n = 5), 80 mg/kg gefitinib (n = 6), 2.35 g/kg lyophilised powder of YYJDD (n = 6) or combination treatment (n = 6) by gavage for 4 weeks and submitted to TUNEL, immunohistochemistry, and western-blot.

Results

In vitro, gefitinib (IC₅₀: 20.68 ± 2.06 μM) and YYJDD (IC₅₀: 6.6 ± 0.21 mg/mL) acted in a moderate synergistic way. Combination treatment inhibited cell viability from 100% to 25.66%. Compared to gefitinb (33.23 ± 3.99%), cell apoptosis was increased with combination treatment (54.11 ± 7.32%), accompanied by down-regulation of the PI3K/Akt. LY294002 further inhibited cell viability, increased apoptosis, and down-regulated p-Akt/Akt. In vivo, the tumour sizes in the combination group (1165.13 ± 157.79 mm³) were smaller than gefitinib alone (1630.66 ± 208.30 mm³). The positive rate of TUNEL staining was increased by combination treatment (22.33 ± 2.75%) versus gefitinib (7.37 ± 0.87%), while the PI3K/Akt was down-regulated.

Discussion and conclusion

YYJDD has potential to overcome gefitinib resistance. Future investigations should be focussed on its specific targets.

中文翻译：

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养阴解毒汤通过下调PI3K/Akt信号通路克服非小细胞肺癌吉非替尼耐药

摘要

语境

在我们的临床实践中，养阴解毒汤（YYJDD）被用来提高吉非替尼的疗效，但其作用机制尚不清楚。

客观的

本研究探讨了 YYJDD 是否可以逆转吉非替尼耐药性。

材料和方法

将 H1975 细胞暴露于对照、10 μM 吉非替尼、3.2 mg/mL YYJDD 或联合治疗。MTT 在 0-96 小时内检测细胞活力。在 24 小时通过流式细胞术和蛋白质印迹检测细胞凋亡和 PI3K/Akt 蛋白。LY294002 用于进一步确定 PI3K/Akt 的作用。23只BALB/c裸鼠移植接受生理盐水（n  = 5）、80 mg/kg吉非替尼（n  = 6）、2.35 g/kg YYJDD冻干粉（n  = 6）或联合治疗（n  = 6）灌胃4周并提交给TUNEL、免疫组化和western-blot。

结果

在体外，吉非替尼 (IC ₅₀ : 20.68 ± 2.06 μM) 和 YYJDD (IC ₅₀ : 6.6 ± 0.21 mg/mL) 以适度的协同方式发挥作用。联合治疗将细胞活力从 100% 抑制到 25.66%。与吉非替尼 (33.23 ± 3.99%) 相比，联合治疗增加细胞凋亡 (54.11 ± 7.32%)，同时下调 PI3K/Akt。LY294002 进一步抑制细胞活力，增加细胞凋亡，并下调 p-Akt/Akt。在体内，联合组的肿瘤大小（1165.13 ± 157.79 mm ³）小于单独的吉非替尼（1630.66 ± 208.30 mm ^{3 ）}）。与吉非替尼（7.37±0.87%）相比，联合治疗增加了TUNEL染色的阳性率（22.33±2.75%），同时下调了PI3K/Akt。

讨论与结论

YYJDD 具有克服吉非替尼耐药性的潜力。未来的调查应该集中在它的具体目标上。