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Sodium butyrate induces autophagy in colorectal cancer cells through LKB1/AMPK signaling

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Abstract

Butyrate is produced by the fermentation of undigested dietary fibers and acts as the promising candidate for cancer treatment. However, the mechanism underlying sodium butyrate (NaB)-induced autophagy in colorectal cancer is not yet completely understood. The expressions of LC3-II protein and mRNA were detected by western blot and quantitative RT-PCR in colorectal cancer (CRC) cell lines HCT-116 and HT-29, respectively. Autolysosome formation was observed by transmission electron microscope. AMPK and LKB1 were inhibited by chemical inhibitor or siRNAs and confirmed by western blot. NaB elevated the protein and mRNA expressions of LC3 in a dose-dependent manner. NaB treatment increased the formation of autolysosome and expression of phosphorylated liver kinase B1 (LKB1), AMP-activated protein kinase (AMPK), and acetyl-CoA carboxylase (ACC). Treatment with compound C (an inhibitor of AMPK) and siRNA-mediated knockdown of AMPK and LKB1 significantly attenuated NaB-induced autophagy in CRC cells. Collectively, these findings indicated that LKB1 and AMPK are critical for NaB-mediated autophagy and may act as the novel targets for colorectal cancer therapy in the future.

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Funding

This study was supported by the grants from National Natural Science Foundation of China (No. 81773429 and 81202204), Guangdong National Natural Science Foundation (No. S2012010009467), Project for Excellent Young University Teacher awarded to Southern Medical University.

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Authors and Affiliations

  1. Department of Nutrition and Food Hygiene, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, No.1023 South Sha-Tai Rd, Guangzhou, 510515, People’s Republic of China

    Shunli Luo, Ziyin Li, Lianzhi Mao & Suxia Sun

  2. School of Laboratory Medicine, Hunan University of Medicine, Huaihua, 418000, People’s Republic of China

    Shunli Luo

  3. Guangzhou Customs District, No.66, Huacheng Avenue, Zhujiang Xincheng, Guangzhou, 510623, People’s Republic of China

    Siqiang Chen

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  1. Shunli Luo
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Correspondence to Suxia Sun.

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Shunli Luo and Ziyin Li are co-first authors.

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Supplementary Fig 1

Knockdown the expression of AMPKα. HCT-116 (A) and HT-29 (B) cells were transfected with negative control (NC) or AMPKα specific siRNAs for 72 h. Protein expression of AMPKα was examined by western blot. mRNA levels of AMPKα in HCT-116 (C) and HT-29 (D) cells were determined using real-time RT-qPCR. Expression levels of AMPKα, GAPDH and the ratio of AMPKα to GAPDH were calculated. *P < 0.05, **P < 0.01 compared to the NC group. (PNG 279 kb)

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Supplementary Fig 2

Knockdown the expression of LKB1. HCT-116 (A) and HT-29 (B) cells were transfected with negative control (NC) or LKB1 specific siRNAs for 72 h. Protein expression of LKB1 was examined by western blot. mRNA levels ofLKB1 in HCT-116 (C) and HT-29 (D) cells were determined using real-time RT-qPCR. Expression levels of LKB1, GAPDH and the ratio of LKB1 to GAPDH were calculated. *P < 0.05, **P < 0.01, ***P < 0.001 compared to the NC group. (PNG 281 kb)

High Resolution Image (TIF 3712 kb)

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Luo, S., Li, Z., Mao, L. et al. Sodium butyrate induces autophagy in colorectal cancer cells through LKB1/AMPK signaling. J Physiol Biochem 75, 53–63 (2019). https://doi.org/10.1007/s13105-018-0651-z

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  • DOI: https://doi.org/10.1007/s13105-018-0651-z

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