Suppression of the Proliferation of Huh7 Hepatoma Cells Involving the Downregulation of Mutant p53 Protein and Inactivation of the STAT 3 Pathway with Ailanthoidol

Tsui-Hwa Tseng; Chau-Jong Wang; Yean-Jang Lee; Yi-Chia Shao; Chien-Heng Shen; Ko-Chao Lee; Shui-Yi Tung; Hsing-Chun Kuo

doi:10.3390/ijms23095102

Suppression of the Proliferation of Huh7 Hepatoma Cells Involving the Downregulation of Mutant p53 Protein and Inactivation of the STAT 3 Pathway with Ailanthoidol

Int J Mol Sci. 2022 May 4;23(9):5102. doi: 10.3390/ijms23095102.

Authors

Tsui-Hwa Tseng^{1

2}, Chau-Jong Wang^{3

4}, Yean-Jang Lee⁵, Yi-Chia Shao¹, Chien-Heng Shen⁶, Ko-Chao Lee⁷, Shui-Yi Tung^{6

8}, Hsing-Chun Kuo^{9

10

11

12}

Affiliations

¹ Department of Medical Applied Chemistry, Chung Shan Medical University, Taichung 40201, Taiwan.
² Department of Medical Education, Chung Shan Medical University Hospital, Taichung 40201, Taiwan.
³ Department of Health Diet and Industry Management, Chung Shan Medical University, Taichung 40201, Taiwan.
⁴ Department of Medical Research, Chung Shan Medical University Hospital, Taichung 40201, Taiwan.
⁵ Department of Chemistry, National Changhua University of Education, Changhua 50007, Taiwan.
⁶ Department of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Chiayi 61363, Taiwan.
⁷ Division of Colorectal Surgery, Department of Surgery, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
⁸ Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Chiayi 61363, Taiwan.
⁹ Division of Basic Medical Sciences, Department of Nursing, Chang Gung University of Science and Technology, Chiayi 61363, Taiwan.
¹⁰ Research Fellow, Chang Gung Memorial Hospital, Chiayi 61363, Taiwan.
¹¹ Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, Taiwan.
¹² Chronic Diseases and Health Promotion Research Center, Chang Gung University of Science and Technology, Chiayi 61363, Taiwan.

Abstract

Ailanthoidol (ATD) has been isolated from the barks of Zanthoxylum ailanthoides and displays anti-inflammatory, antioxidant, antiadipogenic, and antitumor promotion activities. Recently, we found that ATD suppressed TGF-β1-induced migration and invasion of HepG2 cells. In this report, we found that ATD exhibited more potent cytotoxicity in Huh7 hepatoma cells (mutant p53: Y220C) than in HepG2 cells (wild-type p53). A trypan blue dye exclusion assay and colony assay showed ATD inhibited the growth of Huh7 cells. ATD also induced G1 arrest and reduced the expression of cyclin D1 and CDK2. Flow cytometry analysis with Annexin-V/PI staining demonstrated that ATD induced significant apoptosis in Huh7 cells. Moreover, ATD increased the expression of cleaved PARP and Bax and decreased the expression of procaspase 3/8 and Bcl-xL/Bcl-2. In addition, ATD decreased the expression of mutant p53 protein (mutp53), which is associated with cell proliferation with the exploration of p53 siRNA transfection. Furthermore, ATD suppressed the phosphorylation of the signal transducer and activator of transcription 3 (STAT3) and the expression of mevalonate kinase (MVK). Consistent with ATD, the administration of S3I201 (STAT 3 inhibitor) reduced the expression of Bcl-2/Bcl-xL, cyclin D1, mutp53, and MVK. These results demonstrated ATD's selectivity against mutp53 hepatoma cells involving the downregulation of mutp53 and inactivation of STAT3.

Keywords: STAT3; ailanthoidol; apoptosis; cell cycle; hepatoma; mutant p53.

MeSH terms

Aminosalicylic Acids
Apoptosis / physiology
Benzenesulfonates
Benzofurans* / pharmacology
Carcinoma, Hepatocellular* / genetics
Carcinoma, Hepatocellular* / metabolism
Cell Line, Tumor
Cell Proliferation / drug effects
Cyclin D1 / metabolism
Down-Regulation
Humans
Liver Neoplasms* / genetics
Liver Neoplasms* / metabolism
Mutant Proteins / metabolism
Proto-Oncogene Proteins c-bcl-2 / metabolism
STAT3 Transcription Factor / metabolism
Signal Transduction
Tumor Suppressor Protein p53 / metabolism

Substances

Aminosalicylic Acids
Benzenesulfonates
Benzofurans
Mutant Proteins
NSC 74859
Proto-Oncogene Proteins c-bcl-2
STAT3 Transcription Factor
Tumor Suppressor Protein p53
ailanthoidol
Cyclin D1

Grants and funding

This research was supported by the Ministry of Science and Technology Grant (MOST 108-2320-B-040-016-MY3, MOST 110-2320-B-255-005-MY3), Taiwan. Digital Image Analyzer was performed in the Instrument Center of Chung Shan Medical University, which was supported by the Ministry of Science and Technology, Ministry of Education, and Chung Shan Medical University (CSMU 108-OM-A-114), Taiwan.