Acetylshikonin exerts anti-tumor effects on non-small cell lung cancer through dual inhibition of STAT3 and EGFR

Yemeng Tang; Yanmao Wang; Xian Wang; Zhucheng Zhao; Haijian Cai; Mengyao Xie; Xintong Jiang; Luyao Zhang; Jiayun Cheng; Lehe Yang; Liangxing Wang; Chengguang Zhao; Xiaoying Huang

doi:10.1016/j.phymed.2022.154109

Acetylshikonin exerts anti-tumor effects on non-small cell lung cancer through dual inhibition of STAT3 and EGFR

Phytomedicine. 2022 Jul:101:154109. doi: 10.1016/j.phymed.2022.154109. Epub 2022 Apr 30.

Authors

Affiliations

¹ Key Laboratory of Heart and Lung, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
² Key Laboratory of Heart and Lung, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China. Electronic address: wangliangxing@wzhospital.cn.
³ Key Laboratory of Heart and Lung, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; School of Pharmaceutical Sciences, Wenzhou Medical University, Building 11, Chashan Street, University Town, Wenzhou, Zhejiang 325035, China. Electronic address: zhaochengguang@wmu.edu.cn.
⁴ Key Laboratory of Heart and Lung, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China. Electronic address: huangxiaoying@wzhospital.cn.

PMID: 35526322
DOI: 10.1016/j.phymed.2022.154109

Abstract

Background: Lung cancer is one of the most common types of malignant tumor. It has one of the highest morbidity and mortality rates worldwide, and approximately 85% of cases are non-small cell lung cancer (NSCLC). Clinically, several EGFR inhibitors have been used to treat NSCLC, but resistance can develop. Studies have shown that cross talk between signal transducer and activator of transcription 3 (STAT3) and epidermal growth factor receptor (EGFR) can mediate drug resistance. Acetylshikonin has obvious antitumor effects, but the mechanism of action is still unclear.

Purpose: To analyze the antitumor activity of acetylshikonin in lung cancer and clarify its molecular mechanism.

Methods: Methyl thiazolyl tetrazolium (MTT), colony formation and 5-ethynyl-2'-deoxyuridine (EDU) assays were performed to examine the effects of acetylshikonin in inhibiting the proliferation of NSCLC cells (PC-9, H1975 and A549). Scratch wound and transwell assays were used to evaluate the migration and invasion of NSCLC cells. Flow cytometry was employed to determine whether acetylshikonin could induce apoptosis. Proteome sequencing was used to identify the targets of acetylshikonin. Immunofluorescence staining and western blotting were utilized to verify the inhibition of STAT3 and EGFR phosphorylation. A xenotransplantation model was established to evaluate the efficacy of acetylshikonin in nude mice.

Results: Our data demonstrated that acetylshikonin significantly decreased the survival rate of human NSCLC cells, increased the apoptotic rate and inhibited cell migration dose-dependently. Immunofluorescence staining and western blotting analyses revealed that acetylshikonin inhibited EGFR and STAT3 pathways. Acetylshikonin also inhibited tumor growth in a xenograft model better than inhibitors of EGFR and STAT3.

Conclusion: Acetylshikonin has anti-cancer effects on NSCLC cells by inhibiting EGFR and STAT3, indicating that acetylshikonin may be a new antitumor drug to treat NSCLC.

Keywords: Acetylshikonin; Double target; EGFR; Non-small cell lung cancer; STAT3.

MeSH terms

Animals
Anthraquinones
Apoptosis
Carcinoma, Non-Small-Cell Lung* / pathology
Cell Line, Tumor
Cell Proliferation
ErbB Receptors / metabolism
Humans
Lung Neoplasms* / pathology
Mice
Mice, Nude
STAT3 Transcription Factor / metabolism
Xenograft Model Antitumor Assays

Substances

Anthraquinones
STAT3 Transcription Factor
STAT3 protein, human
EGFR protein, human
ErbB Receptors
acetylshikonin