Sweet Wormwood and Tortoise Shell Decoction (Thanh Hao Miet Giap Thang) Induces DNA Damage, S-Phase Arrest, and Apoptosis in MCF-7 Cells via ATR-CHK1 Signaling Pathway.

Hoang Tam Nguyen Thai,Thuy Vy Nguyen,Thuy Duong Ho Huynh,Jun Yan Tao

doi:10.1155/2022/2358290

Abstract

Introduction Sweet wormwood and tortoise shell decoction, Thanh Hao Miet Giap Thang (THMGT) in Vietnamese, a traditional formula composed of five ingredients, is used in complementary care in Vietnam for patients who underwent conventional cancer treatment. To expand the clinical use and explore novel functions of THMGT, this study was conducted to investigate the effect of THMGT in terms of antiproliferative activity and selective cytotoxicity toward human breast cancer cells MCF-7. Methods Cytotoxicity of THMGT against human breast cancer cells MCF-7 and primary fibroblasts from a heathy donor were studied using sulforhodamine B (SRB) assay. Flow cytometry analysis, immunofluorescence, and western blotting were also performed to elucidate underlying mechanisms of THMGT action. Results The SRB assay on treated MCF-7 cells and primary fibroblasts from a heathy donor indicated selective cytotoxicity of THMGT with a selective index of 3.92. Annexin V/PI staining and flow cytometric analysis on stained MCF-7 cells showed that the THMGT-treated cells were arrested at the S phase and subsequently underwent apoptosis. Western blot analysis showed an upregulation of γ-H2AX, increased protein levels of phosphorylated CHK1, TP53, and phosphorylated TP53 in a time-dependent manner, and a downregulated expression of ATR and MDM2. Conclusion These results suggested DNA damaging effect and ATR-CHK1-mediated cell cycle arrest of THMGT on MCF-7 cells resulting in apoptosis induction.

Highlights

Sweet wormwood and tortoise shell decoction, anh Hao Miet Giap ang (THMGT) in Vietnamese, a traditional formula composed of five ingredients, is used in complementary care in Vietnam for patients who underwent conventional cancer treatment
Our results showed that High-Performance Liquid Chromatography (HPLC) chromatograms of THMGT decoctions have similar patterns, suggesting reproducibility in the preparation technology and the quality of the medicinal materials (Figure 1(c))
We assessed the selective cytotoxicity of THMGT through calculation of its selective index (SI). e SI value was determined as the ratio between IC50 values of the formula for human primary fibroblasts and for human breast cancer cells MCF-7. e fibroblasts and MCF-7 cells were treated with the same serial dilution of THMGT and subjected to sulforhodamine B (SRB) assay to determine IC50 values (Figure 2)

Summary

Introduction

Sweet wormwood and tortoise shell decoction, anh Hao Miet Giap ang (THMGT) in Vietnamese, a traditional formula composed of five ingredients, is used in complementary care in Vietnam for patients who underwent conventional cancer treatment. Cytotoxicity of THMGTagainst human breast cancer cells MCF-7 and primary fibroblasts from a heathy donor were studied using sulforhodamine B (SRB) assay. E SRB assay on treated MCF-7 cells and primary fibroblasts from a heathy donor indicated selective cytotoxicity of THMGT with a selective index of 3.92. Annexin V/PI staining and flow cytometric analysis on stained MCF-7 cells showed that the THMGT-treated cells were arrested at the S phase and subsequently underwent apoptosis. E sweet wormwood and tortoise shell decoction (Qing Hao Bie Jia Tang in Chinese, anh Hao Miet Giap ang (THMGT) in Vietnamese) is a traditional Chinese medicine (TCM) formula, traditionally used to treat late-stage chronic fever diseases. Carapax Trionycis, the tortoise shell, was obtained from Tryonix sinensis Wiegmann. e substance alone, or combined into multi-ingredient formulae, was used for the treatment of endometriosis; it inhibits the proliferation and induces apoptosis in hepatic stellate cells

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Conclusion