TKH-03在体内外逆转A2780T细胞对紫杉醇的耐药性研究

TKH-03 reverses paclitaxel resistance in A2780T cells in vitro and in vivo

目的:体内外研究TKH-03对ABCB1介导的卵巢癌耐药的影响及作用机制,探讨TKH-03能否作为耐药逆转剂。方法:采用细胞克隆实验检测TKH-03在体外对A2780T细胞的克隆、增殖影响。通过Ki-67免疫组化染色观察TKH-03在体内对A2780T细胞的影响,同时进行初步安全性评价。分子对接分析,探究与维拉帕米相比,TKH-03与ABCB1分子模型的相互作用情况。结果:细胞克隆实验显示,TKH-03与紫杉醇联用对A2780T细胞的体外增殖能力具有较强的抑制作用。体内抗肿瘤研究显示,TKH-03能增强紫杉醇介导的肿瘤组织细胞凋亡,差异有统计学意义(P<0.01)。TKH-03联合紫杉醇对荷瘤鼠的肝肾功能无明显影响,未发生明显毒副作用。分子对接结果显示,TKH-03与ABCB1通过氢键和疏水作用紧密结合。与ABCB1底物维拉帕米相比,TKH-03与ABCB1的结合更稳定。结论:TKH-03在体内外逆转A2780T细胞对紫杉醇的耐药性,且与紫杉醇联用具有良好的安全性,其逆转耐药机制与ABCB1靶点有关。

Objective:To study the effect of TKH-03 on ABCB1-mediated ovarian cancer resistance and its mechanism in vitro and in vivo,and to explore whether TKH-03 can be used as a drug resistance reversal agent.Methods:Cell cloning experiment was conducted to investigate the effect of TKH-03 on the cloning and proliferation of A2780T in vitro.Ki-67 immunohistochemical staining was used to observe the effect of TKH-03 on A2780T cells in vivo,and preliminary safety evaluation was conducted at the same time.Molecular docking analysis was conducted to explore the interaction between TKH-03 and ABCB1 molecular model compared with commercially available verapamil.Results:The cell cloning experiment showed that the combination of TKH-03 and paclitaxel had a strong inhibitory effect on the proliferation of A2780T cells in vitro.In vivo anti-tumor studies showed that TKH-03 could enhance the apoptosis of tumor tissue cells mediated by paclitaxel,and there was a significant difference.The indexes of liver and kidney function were all within the normal range,and there was no significant difference.No obvious toxic or side effects occurred.Molecular docking results showed that TKH-03 was closely bound to ABCB1 through hydrogen bonding and hydrophobic interaction.Compared with verapamil,the substrate of ABCB1,the binding of TKH-03 to ABCB1 is more stable.Conclusion:TKH-03 can reverse paclitaxel resistance in A2780T cells in vitro and in vivo,and has good safety when combined with paclitaxel.The mechanism of reversing resistance is related to ABCB1.