title: Multiple effects of Xihuang pill aqueous extract on the Hs578T triple-negative breast cancer cell line.
abstract: The management of triple-negative breast cancer (TNBC) is challenging due to the aggressive behavior, lack of therapeutic options and relatively poor prognosis. Xihuang pill (XHP) is a well-known Traditional Chinese Medicine with anticancer activity. The aim of the present study was to investigate whether the aqueous extract of XHP (AEXHP) has anti-proliferative activity against the Hs578T TNBC cell line, and to elucidate its molecular mechanisms of action. First, an MTT assay was used to evaluate the anti-proliferative activity of AEXHP on the Hs578T cell line; furthermore, the cell cycle distribution, mitochondrial membrane potential and apoptotic rate were determined by flow cytometry, and western blot analysis was used to assess the expression of apoptosis and cell cycle regulatory proteins to investigate the mechanisms of action. The results revealed that the cell viability was significantly inhibited by AEXHP in a dose- and time-dependent manner. Apoptosis and mitochondrial membrane potential loss were detected, and after treatment with 4, 8 and 12 mg/ml AEXHP for 24 h, cleaved caspase-3 was 1.70-, 1.81- and 1.84-fold of that of the control, while procaspase-3, procaspase-8, cleaved caspase-8, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) and the Bcl-2/Bax ratio were not significantly affected. Cell cycle analysis revealed that treatment with AEXHP led to S-phase arrest of Hs578T cells. Furthermore, AEXHP treatment resulted in decreased expression of cyclin A and cyclin dependent kinase 2 (CDK2), and increased expression of cyclin E and p21Cip1, as compared to the control group. In conclusion, the viability of Hs578T cells was significantly inhibited by AEXHP in a dose- and time-dependent manner, the likely mechanisms of which being induction of apoptosis, probably via the intrinsic, Bcl-2-independent pathway, and cell cycle arrest in S phase due to decreased expression of cyclin A and CDK2, and increased expression of cyclin E and p21Cip1.
authors.affiliation: Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China; Department of Integration of Chinese and Western Medicine, Peking University School of Oncology, Beijing 100142, P.R. China.
authors.forename: Wenxian
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China; Department of Integration of Chinese and Western Medicine, Peking University School of Oncology, Beijing 100142, P.R. China.
authors.forename: Shuyan
authors.initials: S
authors.lastname: Han
authors.affiliation: Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China; Department of Integration of Chinese and Western Medicine, Peking University School of Oncology, Beijing 100142, P.R. China.
authors.forename: Shantong
authors.initials: S
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China; Department of Integration of Chinese and Western Medicine, Peking University School of Oncology, Beijing 100142, P.R. China.
authors.forename: Lina
authors.initials: L
authors.lastname: Pang
authors.affiliation: Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China; Department of Integration of Chinese and Western Medicine, Peking University School of Oncology, Beijing 100142, P.R. China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Li
authors.affiliation: Central Laboratory, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.
authors.forename: Xijuan
authors.initials: X
authors.lastname: Liu
authors.affiliation: Central Laboratory, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.
authors.forename: Minhua
authors.initials: M
authors.lastname: Cao
authors.affiliation: Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China; Department of Integration of Chinese and Western Medicine, Peking University School of Oncology, Beijing 100142, P.R. China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Li
keywords: Hs578T; Xihuang pill; anti-proliferative activity; apoptosis; aqueous extract; cell cycle; triple-negative breast cancer