N-acetylcysteine reduces artesunate-induced pancreatic carcinoma cell death by activating protective autophagy via the AMPK/mTOR pathway
- VernacularTitle:N-乙酰-L-半胱氨酸通过AMPK/mTOR途径激活保护性自噬降低青蒿琥酯诱导的胰腺癌细胞死亡
- Author:
Zijing REN
1
;
Hongxia XU
1
;
Xingyue LI
2
;
Yue WANG
2
;
Jiajia MA
2
;
Peiyang ZHOU
1
Author Information
- Publication Type:Journal Article
- Keywords: artesunate; pancreatic carcinoma; reactive oxygen species （ROS）; autophagy; AMPK/mTOR
- From: Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(3):354-360
- CountryChina
- Language:Chinese
- Abstract: 【Objective】 In this study， reactive oxygen species （ROS） scavenger N-acetyl-L-cysteine （NAC） was used to explore the inhibitory effect and mechanism of artesunate （ART） on pancreatic carcinoma （PC） cells. 【Methods】 Different concentrations of ART interfered with 3 PC cell lines CFPAC-1， Capan-2 and BxPC3. Cell viability was measured by CCK8； cell migration ability was measured by Transwell method， and the expressions of migration-related proteins E-cadherin， N-cadherin and Vimentin were measured by Western blotting. ROS probe DCFH-DA was used to measure intracellular ROS； LC3 cell immunofluorescence （IF） was used to detect the formation of intracellular autophagosomes. After adding NAC or autophagy inhibitor 3-MA， the cell viability was tested again by CCK8， and the expressions of p-AMPK/ AMPK， p-mTOR/mTOR， p62 and LC3Ⅱ/Ⅰ were detected by Western blotting. 【Results】 ART inhibited the growth of CFPAC-1 and Capan-2 in a time- and dose-dependent manner. After treatment of CFPAC-1 and Capan-2 cells with 200 μmol/L of ART for 48 h， the expression of E-cadherin was upregulated， while N-cadherin and Vimentin were downregulated， and the cell migration ability was significantly reduced. ART significantly upregulated intracellular ROS level and promoted the formation of autophagosomes. NAC could reduce the inhibitory effect of ART on CFPAC-1 and Capan-2 cells， upregulate p-AMPK/AMPK， P62 and LC3Ⅱ/Ⅰ， downregulate the expression of p-mTOR/mTOR， and intensify autophagy. 3-MA could not reverse the inhibitory effect of ART on PC cells. 【Conclusion】 ART is dependent on ROS， but not on autophagy， in exerting an anti-pancreatic carcinoma effect. NAC attenuates the inhibitory effect of ART on PC cells by activating protective autophagy through AMPK/mTOR signaling pathway.
