Regulation of p21, MMP-1, and MDR-1 Expression in Human Colon Carcinoma HT29 Cells by Tian Xian Liquid, a Chinese Medicinal Formula, In Vitro and In Vivo

Abstract

Ethnopharmacological relevance. Tian-Xian liquid (TXL), a commercially available Chinese medicine decoction, has been used as an anticancer dietary agent for more than 10 years without reported side effects. Aim of the study. The safety and quality consistency of TXL and its mechanisms of action on antiproliferation, antimetastasis, and reversion of multidrug resistance (MDR) regimens were explored. Materials and methods. In this study, an atomic absorption spectrophotometer and reversed phase high performance liquid chromatography with photodiode array detection (HPLC-DAD) were used to evaluate the main toxic elements and the quality consistency among different batches of TXL extracts, respectively. HT29 human colon cancer cell line and tumor-bearing nude mice were used. TXL was provided by China-Japan Feida Union Company Limited. The effect of TXL on in vitro proliferation of HT29 human colon cancer cell line was examined. The percentages of treated cells distributed in different phases of the cell cycles were analyzed by flow cytometry. Antiproliferative effect after treatment with TXL was assessed by determination of the protein levels of p21, cyclinD1, PCNA, and cdk-2, which are the key regulators for cell cycle progression. Meanwhile, the protein levels of MMP-1 and MDR-1 (multidrug resistance protein-1) were also determined to assess the effect of TXL on antimetastasis and reversion of MDR regimen, respectively. Results. The contents of main toxic elements were lower in TXL extract compared with the standard set by the Department of Health of the Government of Hong Kong Special Administrative Region (SAR). Our HPLC results showed that the relative standard deviations of the amount of the 5 standards were less than 5% in different batches of TXL. Immunoblotting analysis revealed a dramatic induction of cyclin kinase inhibitor p21 as well as an inhibition of cyclinD1, PCNA, and cdk-2 in the TXL-treated in vitro models, thereby, impeding cell progression from G1/S phase. Results obtained from the in vivo study also demonstrated that TXL upregulated the protein level of p21 and downregulated the protein levels of MMP-1 and MDR-1. Conclusions. Results obtained from the present investigation not only demonstrate the safety and quality of TXL extract but also demonstrate that TXL possesses antiproliferative and antimetastatic activities and brings about reversion of MDR on HT29 cell and on xenografted tissue in tumor-implanted nude mice.

Differential effects of anti-metastatic mechanism of Tian-Xian liquid (TXL) and its bioactive fractions on human colorectal cancer models

“…1). The inhibitory effect of TXL on HT-29 cells was consistent with our previous publications Sze et al, 2011). This provides evidence that the Chinese medicinal formulation, TXL and its bioactive fractions could specifically kill human cancer cells.…”

“…The aqueous, ethyl acetate and butanol fractions were evaporated to yield 60, 8 and 30 g, respectively for subsequent experiments. The quality control of the fractions was performed as described in our previous publication (Sze et al, 2011).…”

“…These herbal medications exhibit anti-neoplastic properties and have long been applied for cancer treatment (Jonas, 1998;Yuan and Lin, 2000;Gao et al, 2007;et al, 2011). Here we further investigate and compare the antimetastatic effects of TXL and its fractions in human colorectal carcinoma cells in vitro and in vivo in order to demonstrate the multi-targeting characteristics of TXL in colorectal cancer.…”

An In Vitro and In Vivo Investigation of the Antimetastatic Effects of a Chinese Medicinal Decoction, Erxian Decoction, on Human Ovarian Cancer Models

“…25 The proteins were extracted from EXDtreated OVCA429 cells, untreated, and EXD-treated excised xenografts. Fifteen micrograms of the denatured protein from each sample was separated and transferred to a polyvinylene difluoride membrane (Pall, Bio-Gene, Pensacola, FL).…”

Beta-catenin mediates the apoptosis induction effect of celastrol in HT29 cells

“…In addition, synchronous colorectal cancers have also risen globally, challenging the current treatment strategies and adversely affecting the overall prognosis of colorectal cancer (Yang et al, 2011b). A human colorectal cancer cell line, HT29 has been commonly used as an in vitro model system to investigate the molecular mechanisms of apoptosis (Lan et al, 2010;Reyes-Zurita et al, 2009Singh et al, 2009;Sze et al, 2010). The process of apoptosis is controlled by a diverse range of cell signals.…”