Antitumor effects of Xi Huang pills on MDA‑MB‑231 cells in vitro and in vivo

Zheng,Wenxian; Han,Shuyan; Jiang,Shantong; He,Xiran; Li,Xiaohong; Ding,Huirong; Cao,Minhua; Li,Pingping

doi:10.3892/mmr.2018.9203

Antitumor effects of Xi Huang pills on MDA‑MB‑231 cells in vitro and in vivo

Authors:
- Wenxian Zheng
- Shuyan Han
- Shantong Jiang
- Xiran He
- Xiaohong Li
- Huirong Ding
- Minhua Cao
- Pingping Li
View Affiliations

Affiliations: Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Central Laboratory, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China
Published online on: June 21, 2018 https://doi.org/10.3892/mmr.2018.9203
Pages: 2068-2078
Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The management of patients with triple‑negative breast cancer is challenging due to the lack of effective therapeutic options, aggressive behavior and relatively poor prognosis. Xi Huang pills (XHP) are a well‑known traditional Chinese medicine that demonstrate anticancer activities. The aim of the present study was to investigate the antitumor effects of XHP on MDA‑MB‑231 cells in vitro and in vivo, and its potential underlying molecular mechanisms. In the present study, an MTT assay was used to evaluate the antiproliferative activity of XHP on MDA‑MB‑231 cells. In order to investigate the effects further, cell cycle distribution, apoptosis and mitochondrial membrane potential assays were performed, as well as western blot analyses. In addition, a tumor xenograft model was employed to investigate the effects of XHP in vivo. The results of the MTT assay demonstrated that the viability of MDA‑MB‑231 cells was markedly inhibited by XHP in a dose‑ and time‑dependent manner. The inhibitory effect of XHP on the viability of MDA‑MB‑231 cells was greater when compared with MCF‑10A cells. An increase in apoptosis and loss of mitochondrial membrane potential was observed following 4, 8 and 12 mg/ml XHP treatment of MDA‑MB‑231 cells. The protein expression levels of cleaved caspase‑3 were increased by 1.62‑, 2.13‑ and 2.19‑fold, respectively, when compared with the untreated controls, whereas no effects on the expression of B‑cell lymphoma 2 (Bcl‑2) or Bcl‑2‑associated X protein (Bax) were observed. The results of the cell cycle distribution assay analysis demonstrated that XHP treatment arrested cells at the G2/M phase. In addition, XHP treatment decreased the expression of cyclin A and increased the expression of p21Cip1. In vivo experiments revealed that XHP inhibited the growth of MDA‑MB‑231 xenograft tumors without body weight loss, and demonstrated similar effects on the protein expression levels of cleaved caspase 3, cyclin A and p21Cip1 as observed in vitro. In conclusion, the viability of MDA‑MB‑231 cells was inhibited by XHP in a dose‑dependent, time‑dependent and cell‑selective manner in vitro, and the potential underlying mechanisms may involve apoptosis and cell cycle arrest at the G2/M phase. XHP may induce apoptosis in MDA‑MB‑231 cells via the intrinsic pathway, which does not involve the Bcl‑2/Bax ratio. G2/M phase arrest may have been due to the integrated action of decreased cyclin A expression and increased p21Cip1 expression. In addition, XHP inhibited the growth of xenograft tumors in the absence of body weight loss in vivo.

View Figures

View References

1	Kolak A, Kamińska M, Sygit K, Budny A, Surdyka D, Kukiełka-Budny B and Burdan F: Primary and secondary prevention of breast cancer. Ann Agric Environ Med. 24:549–553. 2017.PubMed/NCBI
2	Liedtke C, Bernemann C, Kiesel L and Rody A: Genomic profiling in triple-negative breast cancer. Breast Care (Basel). 8:408–413. 2013. View Article : Google Scholar : PubMed/NCBI
3	Bosch A, Eroles P, Zaragoza R, Viña JR and Lluch A: Triple-negative breast cancer: Molecular features, pathogenesis, treatment and current lines of research. Cancer Treat Rev. 36:206–215. 2010. View Article : Google Scholar : PubMed/NCBI
4	Yadav BS, Sharma SC, Chanana P and Jhamb S: Systemic treatment strategies for triple-negative breast cancer. World J Clin Oncol. 5:125–133. 2014. View Article : Google Scholar : PubMed/NCBI
5	Jiao Q, Wu A, Shao G, Peng H, Wang M, Ji S, Liu P and Zhang J: The latest progress in research on triple negative breast cancer (TNBC): Risk factors, possible therapeutic targets and prognostic markers. J Thorac Dis. 6:1329–1335. 2014.PubMed/NCBI
6	Meng Z, Garrett CR, Shen Y, Liu L, Yang P, Huo Y, Zhao Q, Spelman AR, Ng CS, Chang DZ and Cohen L: Prospective randomised evaluation of traditional Chinese medicine combined with chemotherapy: A randomised phase II study of wild toad extract plus gemcitabine in patients with advanced pancreatic adenocarcinomas. Br J Cancer. 107:411–416. 2012. View Article : Google Scholar : PubMed/NCBI
7	Li YC: A Concise Dictionary of Traditional Chinese Medicine. People's Medical Publishing Press; Beijing: pp. 9001979
8	Jin SR, Zhu BD and Qin XH: Comparative study of anti-tumors effects of Xi huang pellet by different processing methods. Shi Zhen Guo Yi Guo Yao. 19:1735–1737. 2008.
9	Pan G, Wang W, Wang L, Zhang F, Yin X, Wang J and Liang R: Anti-breast cancer effects and mechanisms of Xihuang pill on human breast cancer cell lines. J Tradit Chin Med. 33:770–778. 2013. View Article : Google Scholar : PubMed/NCBI
10	You S, Li W and Guan Y: Tunicamycin inhibits colon carcinoma growth and aggressiveness via modulation of the ERK-JNK-mediated AKT/mTOR signaling pathway. Mol Med Rep. 17:4203–4212. 2018.PubMed/NCBI
11	Wang W, Li N, Luo M, Zu Y and Efferth T: Antibacterial activity and anticancer activity of Rosmarinus officinalis L. essential oil compared to that of its main components. Molecules. 17:2704–2713. 2012. View Article : Google Scholar : PubMed/NCBI
12	Fulda S and Debatin KM: Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene. 25:4798–4811. 2006. View Article : Google Scholar : PubMed/NCBI
13	Budihardjo I, Oliver H, Lutter M, Luo X and Wang X: Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol. 15:269–290. 1999. View Article : Google Scholar : PubMed/NCBI
14	Nicholson DW: Caspase structure, proteolytic substrates, and function during apoptotic cell death. Cell Death Differ. 6:1028–1042. 1999. View Article : Google Scholar : PubMed/NCBI
15	Strasser A, Huang DC and Vaux DL: The role of the bcl-2/ced-9 gene family in cancer and general implications of defects in cell death control for tumourigenesis and resistance to chemotherapy. Biochim Biophys Acta. 1333:F151–F178. 1997.PubMed/NCBI
16	Gao N, Budhraja A, Cheng S, Yao H, Zhang Z and Shi X: Induction of apoptosis in human leukemia cells by grape seed extract occurs via activation of c-Jun NH2-terminal kinase. Clin Cancer Res. 15:140–149. 2009. View Article : Google Scholar : PubMed/NCBI
17	Shapiro GI and Harper JW: Anticancer drug targets: Cell cycle and checkpoint control. J Clin Invest. 104:1645–1653. 1999. View Article : Google Scholar : PubMed/NCBI
18	Choi EJ, Oh HM, Wee H, Choi CS, Choi SC, Kim KH, Han WC, Oh TY, Kim SH and Jun CD: Eupatilin exhibits a novel anti-tumor activity through the induction of cell cycle arrest and differentiation of gastric carcinoma AGS cells. Differentiation. 77:412–423. 2009. View Article : Google Scholar : PubMed/NCBI
19	Zheng W, Han S, Jiang S, Pang L, Li X, Liu X, Cao M and Li P: Multiple effects of Xihuang pill aqueous extract on the Hs578T triple-negative breast cancer cell line. Biomed Rep. 5:559–566. 2016. View Article : Google Scholar : PubMed/NCBI
20	Shen JR, Zhu BD, Qin XH and Zhang XS: Antitumous effects of Xi Huang Pellet on diverse human malignant tumor cell strains (MDA-MB-231, SMMC7721, T24, HL-60, A549). J Sichuan Traditional Chin Med. 24:10–13. 2006.
21	Wang M, Meng JY and He SF: Xihuang Pill () induces mesenchymal-epithelial transition and inhibits loss of apicalbasal polarity in colorectal cancer cell through regulating ZEB1-SCRIB loop. Chin J Integr Med. 20:751–757. 2014. View Article : Google Scholar : PubMed/NCBI
22	Xiao H, Qin XH, Lai Y, Shen JR and Lai L: Containing Xihuang Pill drug serum regulates growth of lung cancer stem cells by controlling cyclin D1 of Wnt signaling pathway. Chin J Exp Trad Med Form. 20:172–176. 2014.
23	Xu H, Cui LR and Liu JC: Study on the effects of Xihuang Pill on the expression of Bcl-2 mRNA of mice bearing H22. Modern Preventive Med. 38:2120–2121. 2011.
24	Li LF, Chen RS, Liu XM, Jin QW and Deng XG: Mechanism of Xihuang Pill on inducing liver cancer cell apoptosis. Chin Arch Trad Chin Med. 22:125–126. 2004.
25	Li-na SUN, Jing-yan MENG, Wei WANG, Sen-lin YING, Dan LI, Zuo-ying MA and Yan-tao JIA: Effect of Xihuang Pills on protein expressions of MMP-2 and MMP-9 in human colorectal carcinoma LoVo cell. Tianjin J Traditional Chin Med. 29:378–380. 2012.
26	Wang YY, Ren YZ, Jiao Z, Zeng CQ, Gao WB and Liang WB: The influence of Xihuang Pill on the formation of con tumor-bearing mice. Pharmacol Clin Chin Materia Med. 30:11–13. 2014.
27	Si-feng WANG, Ke-chun LIU, Xi-min WANG, Qiu-xia HE, Xue WANG, Xi-qiang CHEN and Yan-qiang YUAN: Effect of Xihuang Pill on angiogenesis in Zebrafish embryo. Chin J Hospital Pharm. 30:821–823. 2010.
28	Ma Jie, Guan Shuo, Yang Wei, Hu Junxia, Zeng Changqian, Gao Wenbin and Liang Wengbo: Experimental study on the effect of Xihuang Pill ethanol extract on immune function of tumorbearing rats. Pharmacol Clinics of Chin Materia Medica. 29:124–126. 2013.
29	Guan S, Yang W, Hu JX, Ma J, Gao WB and Liang WB: Effect of chloroform extract of Xihuang Pill on the immune clearance function of tumor-bearing rats. Chin J Modern Applied Pharm. 31:144–148. 2014.
30	Yang Wei, Guan Shuo, Hu Junxia, Zeng Changqian, Liang Wenbo, Ma Jie and Gao Wenbin: Experimental study on antitumor effect of volatile oil of Xihuang Pill and its immune mechanism. World Sci Technol-Modern of Trad Chin Med. 16:68–72. 2014.
31	Rebecca SY and Wong: Apoptosis in cancer: From pathogenesis to treatment. J Exp Clin Cancer Res. 30:872011. View Article : Google Scholar : PubMed/NCBI
32	Diaz-Moralli S, Tarrado-Castellarnau M, Miranda A and Cascante M: Targeting cell cycle regulation in cancer therapy. Pharmacol Ther. 138:255–271. 2013. View Article : Google Scholar : PubMed/NCBI
33	Sherr CJ and Roberts JM: CDK inhibitors: Positive and negative regulators of G1-phase progression. Genes Dev. 13:1501–1512. 1999. View Article : Google Scholar : PubMed/NCBI
34	Guo Q, Lin J, Liu R, Gao Y, He S, Xu X, Hua B, Li C, Hou W, Zheng H and Bao Y: Review on the applications and molecular mechanisms of xihuang pill in tumor treatment. Evid Based Complement Alternat Med. 2015:8543072015. View Article : Google Scholar : PubMed/NCBI
35	Yu D and An GY: Clinical effects of xihuang pill combined with chemotherapy in patients with advanced colorectal cancer. Evid Based Complement Alternat Med. 2017:59360862017. View Article : Google Scholar : PubMed/NCBI