The incidence and mortality of lung cancer are the highest among cancer-related deaths. However, the long-term use of currently available cytotoxic drugs can increase genetic alterations in cancer cells and cause drug-resistance, which significantly limits their usage. Since current systemic treatment options are limited, effective chemotherapeutic agents are urgently needed for non-small cell lung cancer (NSCLC) treatment. In this study, we demonstrated that ganoderic acid DM (GA-DM) could increase apoptosis in A549 and NCI-H460 NSCLC cells. GA-DM treatment decreased the protein expression levels of Bcl-2 and increased the expression levels of Bax, cleaved caspase-3 and cleaved PRAP. Furthermore, GA-DM could promote autophagic flux, and the cytotoxic effect against cancer cells of GA-DM was significantly inhibited by targeted suppression of autophagy, suggesting that autophagy contributed to GA-DM-induced cell death in NSCLC. Moreover, GA-DM clearly induced autophagy by inactivating the PI3K/Akt/mTOR pathway. When overexpression of Akt reactivated Akt/mTOR pathway in A549 or NCI-H460 cells, the increase of autophagy related marker LC3B-II and apoptosis related protein cleaved PARP and cleaved caspase 3 and the ration of apoptotic cells by GA-DM was reversed, suggesting that GA-DM promoted autophagy and apoptosis by inhibiting Akt/mTOR pathway-mediated autophagy induction. In conclusion, our study indicated that GA-DM can induce autophagic apoptosis in NSCLC by inhibiting Akt/mTOR activity. (209 words).

中文翻译：

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灵芝酸DM通过抑制PI3K / Akt / mTOR活性诱导非小细胞肺癌细胞自噬凋亡。

在与癌症相关的死亡中，肺癌的发病率和死亡率最高。但是，长期使用目前可用的细胞毒性药物会增加癌细胞的遗传改变并引起耐药性，从而极大地限制了它们的使用。由于当前的全身治疗选择有限，非小细胞肺癌（NSCLC）治疗迫切需要有效的化学治疗剂。在这项研究中，我们证明了灵芝酸DM（GA-DM）可以增加A549和NCI-H460 NSCLC细胞的凋亡。GA-DM处理降低了Bcl-2的蛋白表达水平，并增加了Bax，caspase-3切割和PRAP切割的表达水平。此外，GA-DM可以促进自噬通量，自噬的靶向抑制作用显着抑制了GA-DM对癌细胞的细胞毒作用，提示自噬促进了GA-DM诱导的NSCLC细胞死亡。此外，GA-DM通过使PI3K / Akt / mTOR通路失活而明显诱导自噬。当A549或NCI-H460细胞中Akt的过量表达重新激活了Akt / mTOR途径时，自噬相关标记LC3B-II和凋亡相关蛋白的表达被PARP和caspase 3裂解，GA-DM导致凋亡细胞的比例被逆转，提示GA-DM通过抑制Akt / mTOR途径介导的自噬诱导而促进自噬和细胞凋亡。总之，我们的研究表明，GA-DM可以通过抑制Akt / mTOR活性来诱导NSCLC自噬细胞凋亡。（209个字）。提示自噬促进了GA-DM诱导的NSCLC细胞死亡。此外，GA-DM通过使PI3K / Akt / mTOR通路失活而明显诱导自噬。当A549或NCI-H460细胞中Akt的过量表达重新激活了Akt / mTOR通路时，自噬相关标记LC3B-II和凋亡相关蛋白的表达被PARP和caspase 3裂解，GA-DM导致凋亡细胞的比例被逆转，提示GA-DM通过抑制Akt / mTOR途径介导的自噬诱导而促进自噬和细胞凋亡。总之，我们的研究表明，GA-DM可以通过抑制Akt / mTOR活性来诱导NSCLC自噬细胞凋亡。（209个字）。提示自噬促进了GA-DM诱导的NSCLC细胞死亡。此外，GA-DM通过使PI3K / Akt / mTOR通路失活而明显诱导自噬。当A549或NCI-H460细胞中Akt的过量表达重新激活了Akt / mTOR通路时，自噬相关标记LC3B-II和凋亡相关蛋白的表达被PARP和caspase 3裂解，GA-DM导致凋亡细胞的比例被逆转，提示GA-DM通过抑制Akt / mTOR途径介导的自噬诱导而促进自噬和细胞凋亡。总之，我们的研究表明，GA-DM可以通过抑制Akt / mTOR活性来诱导NSCLC自噬细胞凋亡。（209个字）。GA-DM逆转了自噬相关标志物LC3B-II和凋亡相关蛋白裂解的PARP和caspase 3的裂解以及凋亡细胞的比率，这表明GA-DM通过抑制Akt / mTOR途径介导的自噬促进了自噬和凋亡就职。总之，我们的研究表明，GA-DM可以通过抑制Akt / mTOR活性来诱导NSCLC自噬细胞凋亡。（209个字）。GA-DM逆转了自噬相关标志物LC3B-II和凋亡相关蛋白裂解的PARP和caspase 3的裂解以及凋亡细胞的比率，这表明GA-DM通过抑制Akt / mTOR途径介导的自噬促进了自噬和凋亡就职。总之，我们的研究表明，GA-DM可以通过抑制Akt / mTOR活性来诱导NSCLC自噬细胞凋亡。（209个字）。