阿司匹林通过 PI3K/Akt/Raptor 通路对 PIK3CA 突变型结直肠癌细胞有更好的作用。
Aspirin has a better effect on PIK3CA mutant colorectal cancer cells by PI3K/Akt/Raptor pathway.
机构信息
Department of Colorectal and Anal Surgery, the First Affiliated Hospital, Guangxi Medical University, Nanning, 530021, China.
Department of Gastroenterology, Zhongnan Hospital of Wuhan University, Wuhan, 430000, China.
出版信息
Mol Med. 2020 Jan 30;26(1):14. doi: 10.1186/s10020-020-0139-5.
BACKGROUND
Aspirin, as a non-steroidal anti-inflammatory drug, can improve the survival rate of patients with colorectal cancer, while aspirin is effective in patients with PIK3CA mutant colorectal cancer (CRC). However, the mechanism of aspirin in the treatment of PIK3CA mutated CRC patients remains unclear.
METHODS
In this study, immunohistochemistry was used to detect the expression levels of PI3K and Raptor in colorectal cancer patients with PIK3CA mutation and PIK3CA wild-type patients. To demonstrate that aspirin has a better effect on the CRC of PIK3CA mutations in association with the PI3K/Akt/Raptor pathway, we used aspirin to treat PIK3CA mutant CRC cells (HCT-116 and RKO). Subsequently, the CCK8 assay and flow cytometry assay were used to detect the apoptosis of PIK3CA mutant CRC cells before and after aspirin use. Western blot was used to detect the changes of PI3K/Akt/Raptor-associated protein, autophagy protein microtubule associated protein 1 light chain 3 alpha (MAP1LC3A, LC3), beclin 1 (BECN1) and apoptosis protein BCL2-associated X protein/ BCL2 apoptosis regulator (Bax/Bcl2), Caspase 3 after treatment of CRC cells with PIK3CA mutation by aspirin.
RESULTS
Phosphoinositide-3-kinase (PI3K) and regulatory associated protein of MTOR complex 1 (Raptor) protein expression levels were higher in PIK3CA-mutant patients than in IK3CA wild-type patients. The expression of Bax/Bcl2 increased after treatment indicates that aspirin can induce apoptosis of PIK3CA-mutant CRC cells. The expression level of MAP1LC3 (LC3) in cells increases with the concentration of aspirin demonstrates that aspirin can induce autophagy in CRC cells. After 48 h of treatment with aspirin, the phosphorylation of eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1) and ribosomal protein S6 kinase B1 (S6K1) was reduced, cell proliferation has been inhibited. After treatment with aspirin, as phosphorylation of PI3K and Protein kinase B (PKB, Akt) was decreased, Raptor expression was also decreased.
CONCLUSION
Aspirin can regulate the proliferation, apoptosis and autophagy of CRC cells through the PI3K/Akt/Raptor pathway, affecting PIK3CA-mutant CRC.
背景
阿司匹林作为一种非甾体抗炎药，可以提高结直肠癌患者的生存率，而阿司匹林对 PIK3CA 突变的结直肠癌（CRC）患者有效。然而，阿司匹林治疗 PIK3CA 突变 CRC 患者的机制尚不清楚。
方法
本研究采用免疫组织化学法检测 PIK3CA 突变和 PIK3CA 野生型结直肠癌患者的 PI3K 和 Raptor 表达水平。为了证明阿司匹林与 PI3K/Akt/Raptor 通路相关，对 PIK3CA 突变的 CRC 具有更好的治疗效果，我们用阿司匹林处理 PIK3CA 突变 CRC 细胞（HCT-116 和 RKO）。随后，用 CCK8 法和流式细胞术检测阿司匹林处理前后 PIK3CA 突变 CRC 细胞的凋亡情况。用 Western blot 检测 PI3K/Akt/Raptor 相关蛋白、自噬蛋白微管相关蛋白 1 轻链 3α（MAP1LC3A，LC3）、自噬相关蛋白 Beclin 1（BECN1）和凋亡蛋白 B 细胞淋巴瘤 2 相关 X 蛋白/BCL2 凋亡调节因子（Bax/Bcl2）、Caspase 3 的变化。
结果
PI3K 和 MTOR 复合物 1 调节相关蛋白（Raptor）的磷酸化水平在 PIK3CA 突变患者中高于 PIK3CA 野生型患者。Bax/Bcl2 表达增加表明阿司匹林可以诱导 PIK3CA 突变 CRC 细胞凋亡。细胞中 MAP1LC3（LC3）的表达水平随着阿司匹林浓度的增加而增加，表明阿司匹林可以诱导 CRC 细胞自噬。用阿司匹林处理 48 小时后，真核翻译起始因子 4E 结合蛋白 1（4E-BP1）和核糖体蛋白 S6 激酶 B1（S6K1）的磷酸化减少，细胞增殖受到抑制。用阿司匹林处理后，PI3K 和蛋白激酶 B（PKB，Akt）的磷酸化减少，Raptor 表达也减少。
结论
阿司匹林可以通过 PI3K/Akt/Raptor 通路调节 CRC 细胞的增殖、凋亡和自噬，影响 PIK3CA 突变的 CRC。
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