Anticancer Effect of Isoliquiritigenin on Human Clear Cell Renal Cell Carcinoma 786-O Cells and Molecular Mechanism
10.13422/j.cnki.syfjx.20191122
- VernacularTitle: 异甘草素对人肾透明细胞癌786-O细胞抗癌作用及分子机制
- Author:
Hong XIN
1
;
Peng-ran SUN
1
;
Peng SONG
1
;
Wei XU
1
Author Information
1. The First Affiliated Hospital of Harbin Medical University, Harbin 150001, China
- Publication Type:Research Article
- Keywords:
isoliquiritigenin (ISL);
clear cell renal cell carcinoma;
786-O;
autophagy;
phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(12):83-89
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the anticancer effect of isoliquiritigenin (ISL) on human clear cell renal cell carcinoma 786-O cells, and explore its possible molecular mechanism. Method: Thiazolyl blue tetrazolium bromide (MTT) assay was used to detect effect of ISL (0, 10, 25,50, 75, 100 μmol·L-1) on proliferation of 786-O cells. The effect of ISL on migration and invasion of 786-O cells was detected by cell scratch test and Transwell assay. The autophagy was observed under the fluorescence microscope through acridine orange staining and Ad-GFP-LC3 transfection experiment. Western blot was used to detect the expression of autophagy related protein and analyze the changes of phosphatidylinositol-3-kinase (PI3K)/protein kinase B(Akt)/mammalian target of rapamycin (mTOR) signaling pathway to explore the possible mechanism. Result: MTT results showed that ISL could significantly inhibit the proliferation of 786-O cells in a time-dose dependent manner (P<0.05). The results of scratch and Transwell experiments indicated that ISL could inhibit the migration and invasion of 786-O cells (P<0.05). Acridine orange staining and ad-GFP-LC3 transfection showed that ISL can induce autophagy in 786-O cells. Besides, ISL can induce LC3-Ⅱ, Beclin1, Atg5 protein expressions (P<0.05), and significantly reduce p62 protein expression (P<0.05); autophagy inhibitor 3-methyl adenine (3-MA) can be added to reverse the above phenomena (P<0.05). Meanwhile, ISL can inhibit phosphorylation of Akt, mTOR levels (P<0.05), inhibitor LY294002 and rapamycin (Rap) can be added to significantly re-regulate LC3-Ⅱ, Beclin1, Atg5 protein expressions (P<0.05) and significantly down-regulate p62 protein expression (P<0.05). Conclusion: ISL can inhibit the proliferation, migration and invasion of clear cell renal carcinoma 786-O cells, and induce autophagy by inhibiting the PI3K/Akt/mTOR signaling pathway.
