Laboratory Animal and Comparative Medicine ›› 2020, Vol. 40 ›› Issue (5): 403-.DOI: 10.3969/j.issn.1674-5817.2020.05.007

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Effects of Sodium Oxalate on Autophagy and Apoptosis of Cholangiocarcinoma Cells#br#

HE Dawei1, HE Aolin1, ZHOU Minghui1, YU Yaping2, ZHANG Minghua2   

  1. 1. Clinical Experimental Research Center,  2. Department of Hepatobiliary surgery First People’s Hospital of Kunshan, Kunshan 215300, Jiangsu Province, China
  • Received:2019-12-06 Online:2020-10-15 Published:2020-11-19

Abstract:

Objective    To observe the effects of sodium oxalate, an inhibitor of lactate dehydrogenase A (LDHA), on autophagy and apoptosis of cholangiocarcinoma cell line Hucct-1, and to explore the related mechanisms. Methods    Hucct-1 cells were treated with different concentrations of sodium oxalate. And LDHA siRNA was transfected into Hucct-1 cells by LipofectAMINE 2000 and Opti-MEM medium. Then the cell proliferation was detected by CCK8 assay, the apoptosis was detected by flow cytometry, and the cell migration was detected by scratch healing test. Meanwhile, the expressions of autophagy-related mRNAs and proteins were detected respectively by RT-PCR and Western blotting. And the lactic acid detection kit was used to detect lactate content in cells. Results    Sodium oxalate inhibited the proliferation of Hucct-1 cells in a dose-dependent manner, its  half-inhibitory concentration increased was 50 mmol/L. After Hucct-1 cells were treated with 50 mmol/L sodium oxalate or transfected with LDHA siRNA, the mRNA and protein expression levels of LDHA were decreased (both P<0.01), and the intracellular lactic acid content was decreased (P<0.01). In the meantime, the apoptosis rate was increased (P<0.01), and the cell migration distance was decreased (P<0.01). And the expression levels of autophagy-related gene 7 (ATG7), Beclin-1 and microtubule-associated protein 1 light chain 3Ⅱ (LC-3Ⅱ) were increased (all P<0.01). In addition, the expression of P62 protein was significantly decreased (P<0.01). Conclusion    Sodium oxalate can promote autophagy and induce apoptosis by inhibiting the expression of LDHA in Hucct-1 cells.


Key words: Sodium oxalate, Cholangiocarcinoma, Lactate dehydrogenase A, Autophagy, Apoptosis     

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