Nguyen Thai Hoang Tam, Nguyen Thuy Vy, Ho Huynh Thuy Duong
Department of Genetics, Faculty of Biology and Biotechnology, University of Science, VNUHCM, 227 Nguyen Van Cu Street, District 5, Ho Chi Minh City, Vietnam.
Evid Based Complement Alternat Med. 2022 Mar 5;2022:2358290. doi: 10.1155/2022/2358290. eCollection 2022.
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.
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.
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 -H2AX, increased protein levels of phosphorylated CHK1, TP53, and phosphorylated TP53 in a time-dependent manner, and a downregulated expression of ATR and MDM2.
These results suggested DNA damaging effect and ATR-CHK1-mediated cell cycle arrest of THMGT on MCF-7 cells resulting in apoptosis induction.
青蒿鳖甲汤（越南语为Thanh Hao Miet Giap Thang，简称THMGT）是一种由五味药材组成的传统方剂，在越南用于接受传统癌症治疗患者的辅助护理。为了扩大THMGT的临床应用并探索其新功能，本研究旨在探讨THMGT对人乳腺癌细胞MCF-7的抗增殖活性和选择性细胞毒性作用。
采用磺酰罗丹明B（SRB）法研究THMGT对人乳腺癌细胞MCF-7和健康供体原代成纤维细胞的细胞毒性。还进行了流式细胞术分析、免疫荧光和蛋白质印迹，以阐明THMGT作用的潜在机制。
对处理后的MCF-7细胞和健康供体原代成纤维细胞进行的SRB分析表明，THMGT具有选择性细胞毒性，选择性指数为3.92。对染色的MCF-7细胞进行膜联蛋白V/碘化丙啶染色和流式细胞术分析显示，经THMGT处理的细胞停滞在S期，随后发生凋亡。蛋白质印迹分析显示，γ-H2AX上调，磷酸化CHK1、TP53和磷酸化TP53的蛋白质水平呈时间依赖性增加，ATR和MDM2的表达下调。
这些结果表明，THMGT对MCF-7细胞具有DNA损伤作用以及ATR-CHK1介导的细胞周期阻滞，从而诱导细胞凋亡。
