Echinacoside inhibits hepatocellular carcinoma progression by targeting the miR-30c-5p/FOXD1/KLF12 axis

Article type: Research Article

Authors: Wang, Guoyu^{a; 1} | Han, Yang^{a; 1} | Zhuang, Juhua^a | Mai, Zhongchao^a | Xia, Wei^{b; *} | Ye, Ying^{c; *}

Affiliations: [a] Department of Nuclear Medicine, The Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, China | [b] Department of Nuclear Medicine, Pudong New Area Gongli Hospital, Shanghai, China | [c] Central Laboratory, The Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, China

Correspondence: [*] Corresponding authors: Wei Xia, Department of Nuclear Medicine, Pudong New Area Gongli Hospital, NO. 219, Miaopu Road, Pudong New Area, Shanghai 200135, China. E-mail: [email protected]. Ying Ye, Central Laboratory, The Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, 358 Datong Rd, Pudong New Area, Shanghai 200137, China. E-mail: [email protected].

Note: [1] Contributed equally to this work.

Abstract: BACKGROUND: Hepatocellular carcinoma (HCC) is the third leading cause of cancer-attributed mortality and the primary liver malignancy in the world. Echinacoside is a phenylethanoid glycoside derived from traditional Chinese medicinal herbs which possessed multiple health benefits on humans, including anti-tumor effects. OBJECTIVE: This study aimed to demonstrate the function of echinacoside in HCC progression and the involvement of miR-30c-5p/FOXD1/KLF12 axis. METHODS: The HepG2 cells were treated by different dose of echinacoside, miR-30c-5p mimic, miR-30c-5p inhibitor, and FOXD1 overexpression lentiviruses or siRNA individually or simultaneously. The cell invasion and migration were measured by transwell assay. RNA and protein levels were tested by RT-PCR and western blot, respectively. The regulatory function of miR-30c-5p on Forkhead box D1 (FOXD1), FOXD1 on Krüppel-like factor 12 (KLF12) was tested by luciferase reporter assay or/and ChIP assay. Meanwhile, a liver cancer lung metastasis mice model was used to examine the functions of echinacoside and miR-30c-5p on HCC metastasis in vivo. Moreover, the correlations among miR-30c-5p, FOXD1, KLF12, and HCC prognosis was analyzed using clinical sample and TCGA database. RESULTS: Based on both in vitro and in vivo investigations, we found that echinacoside could inhibit HCC cell migration, invasiveness, and tumor metastasis, and associated with the enhanced miR-30c-5p/FOXD1/KLF12 axis. Furthermore, through analyzing the interactions among intermediate molecules, we revealed that miR-30c-5p, FOXD1, and KLF12üere clinically relevant with each other in HCC patients, correlated with HCC prognosis, and regulated by echinacoside to contribute in the inhibition of HCC progression. CONCLUSIONS: These findings suggest that echinacoside could inhibit HCC progression, and the mechanism related to the enhanced miR-30c-5p/FOXD1/KLF12 axis. Moreover, the abovementioned intermediate molecules might serve as prospective biomarkers for HCC prognosis.

Keywords: Echinacoside, microRNA, FOXD1, KLF12, hepatocellular carcinoma

DOI: 10.3233/THC-241449

Journal: Technology and Health Care, vol. Pre-press, no. Pre-press, pp. 1-16, 2024