Hispidulin induces ER stress-mediated apoptosis in human hepatocellular carcinoma cells in vitro and in vivo by activating AMPK signaling pathway
Authors: Xue-Hong Chen
Yan-Tao Han
Yin-Ping Yuan
Mei Han
Jing Xie
Hui Gao
Zhi-Wu Han
Quan Yuan
Ming-Quan Gao
Kai-Li Liu
Journal: Acta Pharmacologica Sinica (2019)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: AMPK/mTOR; CHOP; Compound C; Z-LEHD-FMK; endoplasmic reticulum stress
Affiliations: Department of Pharmacology, School of Pharmacy, Qingdao University, Qingdao, 266021, China., Department of Pharmacology, School of Pharmacy, Qingdao University, Qingdao, 266021, China. ., Medical College, Qingdao University, Qingdao, 266071, China., Shandong Cancer Hospital Affiliated to Shandong University, Jinan, 250117, China., Shandong Academy of Medical Sciences, Jinan, 250001, China., Qingdao Fifth People's Hospital, Qingdao, 266000, China., The Affiliated Hospital of Qingdao University, Qingdao, 266003, China.

Abstract:  Hispidulin (4',5,7-trihydroxy-6-methoxyflavone) is a phenolic flavonoid isolated from the medicinal plant S. involucrata, which exhibits anti-neoplastic activity against several types of cancer. However, the mechanism underlying its anti-cancer activity against hepatocellular carcinoma (HCC) has not been fully elucidated. In this study, we investigated whether and how hispidulin-induced apoptosis of human HCC cells in vitro and in vivo. We showed that hispidulin (10, 20 μmol/L) dose-dependently inhibited cell growth and promoted apoptosis through mitochondrial apoptosis pathway in human HCC SMMC7721 cells and Huh7 cells. More importantly, we revealed that its pro-apoptotic effects depended on endoplasmic reticulum stress (ERS) and unfolded protein response (UPR), as pretreatment with salubrinal, a selective ERS inhibitor, or shRNA targeting a UPR protein CHOP effectively abrogated hispidulin-induced cell apoptosis. Furthermore, we showed that hispidulin-induced apoptosis was mediated by activation of AMPK/mTOR signaling pathway as pretreatment with Compound C, an AMPK inhibitor, or AMPK-targeting siRNA reversed the pro-apoptotic effect of hispidulin. In HCC xenograft nude mice, administration of hispidulin (25, 50 mg/kg every day, ip, for 27 days) dose-dependently suppressed the tumor growth, accompanied by inducing ERS and apoptosis in tumor tissue. Taken together, our results demonstrate that hispidulin induces ERS-mediated apoptosis in HCC cells via activating the AMPK/mTOR pathway. This study provides new insights into the anti-tumor activity of hispidulin in HCC. 

Retracted: Yes

PubPeer comments:
  Fig. 4 Blocking the ERS signaling pathway reverses hispidulin-induced apoptosis in HCC cells.... d Flow cytometry analysis of apoptotic cells in SMMC7721 and Huh7 cells or CHOP shRNA transfection cells after treatment with hispidulin (20 u03bcM). ![file] Cf. [Yang et al (2019)] ; [Liu et al (2019)] ; etc. ![] , 
  Fig. 6 Hispidulin exhibits anti-tumor activity in vivo through the activation of the ERS signaling pathway. ... b TUNEL assay analysis of apoptotic cell cryostat sections (4 u03bcm sections) of HCC xenograft tumors after treatment with hispidulin. c Immunohistochemistry staining was performed on the cryostat sections (4 u03bcm sections) of HCC xenograft tumors to detect the expression of CHOP, Bcl-2, cleaved caspase-3, and Ki-67 after treatment with hispidulin.  (b) also appears as 7(E),  TUNEL staining showing the number of apoptotic cells in [HCC] tumor sections , from [Pan et al (2018)] *[upper right]*. (c) shares panels with 6(e) of [Zhang et al (2020)] *[lower right]*. ![file] , 
 Retracted [7 September 2021] . ------------------------------------- The Editor-in-Chief has retracted this article. Concerns were raised regarding a number figures, specifically: - Figure 1E has been previously published as Figure 1D in 1. - Figure 1E: the upper left hand panel has been previously published in Figure 7B of 2 The Editor-in-Chief therefore no longer has confidence in the reliability of the data reported in the article. References - [1] Han M, et al. Hispidulin inhibits hepatocellular carcinoma growth and metastasis through AMPK and ERK signaling mediated activation of PPARu03b3. Biomed Pharmacotherapy. 2018;103:272u2013283. https: doi.org 10.1016 j.biopha.2018.04.014 - [2]. Liu K, Gao H, Wang Q, et al. Hispidulin suppresses cell growth and metastasis by targeting PIM1 through JAK2 STAT3 signaling in colorectal cancer. Cancer Sci. 2018;109:1369u20131381. https: doi.org 10.1111 cas.13575 , 
 http: www.most.gov.cn zxgz kycxjs kycxgzdt 202201 t20220127_179206.html Google Translated Date: January 27, 2022 14:44 Source: Ministry of Science and Technology u3000u30009. The paper  Hispidulin induces ER stress-mediated apoptosis in human hepatocellular carcinoma cells in vitro and in vivo by activating AMPK signaling pathway by Gao Hui as the corresponding author and Han Mei as the first author from the School of Pharmacy, Qingdao University, after investigation, the paper There are data fraud and improper signature behavior; Gao Hui is the corresponding author and Han Mei is the first author of the paper  Hispidulin inhibits hepatocellular carcinoma growth and metastasis through AMPK and ERK signaling mediated activation of PPARu03b3 , after investigation, the paper has improper signature behavior . Gao Hui (merged with the other 4 papers) will be dealt with as follows: demerit for 1 year, cancellation of application or application for science and technology projects (special projects, funds, etc.), science and technology awards, and qualifications for science and technology talent titles for 9 years, cancellation as a nomination or recommendation Persons, nominees or recommenders, review experts, etc. have been qualified for 9 years, recorded in the file of scientific research integrity and dishonesty, canceled to participate in this year s evaluation, and the annual evaluation shall not be determined as qualified or above; Han Mei will be dealt with as follows: warning 6 months, cancel the qualifications for applying for scientific research projects, academic awards, honorary titles, etc. for 1 year, and the annual assessment shall not be determined as an excellent grade. u3000u3000(Notification link: https: kjc.qdu.edu.cn info 1023 3176.htm) ,