AKT抑制剂Capivasertib协同增强KRAS G12C抑制剂的抗肿瘤活性

AKT Inhibitor Capivasertib Synergically Enhances The Antitumor Activity of KRAS G12C Inhibitor

关键词:  KRAS G12C  AKT  Capivasertib  抗肿瘤活性  协同作用

Key words:  KRAS G12C inhibitor  AKT inhibitor Capivasertib  Antitumor activity  synergistic effect

摘要:

目的 探索KRAS G12C抑制剂和AKT抑制剂卡帕塞替尼联用对KRAS G12C突变肿瘤细胞的抗肿瘤活性及作用机制。方法 以含有KRAS G12C突变的人胰腺癌细胞MIA-PACA2和人非小细胞肺癌NCI-H358等肿瘤细胞为模型,采用CCK8法、流式细胞术、Western blotting、克隆形成实验等分别考察KRAS G12C抑制剂和卡帕塞替尼单用或联用对细胞增殖抑制、细胞周期凋亡、胞内相关蛋白表达变化以及细胞克隆形成能力的影响。结果 KRAS G12C抑制剂和卡帕塞替尼联用显著增强抑制肿瘤细胞增殖,显著抑制细胞克隆形成,浓度依赖性地下调AKT 通路和ERK通路相关蛋白的活化或表达,进而引起肿瘤细胞发生细胞周期阻滞和凋亡。结论 KRAS G12C抑制剂和卡帕塞替尼联用对KRAS G12C突变的肿瘤细胞具有协同抑制作用,通过诱导细胞周期阻滞和细胞凋亡,发挥对肿瘤细胞显著的抗肿瘤活性。

Abstract:

OBJECTIVE To investigate the antitumor activities and mechanisms of the combination of KRAS G12C inhibitors and AKT inhibitor Capivasertib against KRAS G12- mutant tumor cells. METHODS Taking KRAS G12C mutation cell lines, including human pancreatic cancer cell line MIA-PACA2 and human non-small cell lung cancer NCI-H358 as in vitro model, the CCK8 assay, flow cytometry, Western blotting, colony formation assay were performed to investigate the effects of KRAS G12C inhibitor in combination with capmatinib or alone on cell proliferation inhibition, cell cycle apoptosis, the expression of intracellular related proteins and the ability of colony formation, respectively. RESULTS The combination of KRAS G12C inhibitor and Capivasertib significantly enhanced the inhibition of tumor cell proliferation, cell migration, and colony formation, and down-regulated the activation or expression of AKT pathway and ERK pathway-related proteins in a concentration-dependent manner, thereby inducing cell cycle arrest and apoptosis of tumor cells. CONCLUSION The combination of KRAS G12C inhibitor and Capivasertib has synergistic effects on KRAS G12C mutant tumor cells, and exerts significant antitumor activities through inducing cell cycle arrest and apoptosis.