Yin Yang Gong Ji pill is an ancient formula with antitumor activity against hepatoma cells.

Abstract

Yin Yang Gong Ji pill (YYGJ) is a formula that was used in the Ming Dynasty. This study investigated the effects of YYGJ on HepG2 and MHCC97H hepatoma cells. The effects of YYGJ drug-containing rat serum (YYGJ serum) on cell proliferation and the cell cycle were investigated by a tetrazolium dye-based MTS assay and flow cytometry. Apoptosis was assayed by TUNEL and flow cytometry. E-cadherin, vimentin, c-Myc, Smad4, and MMP2 expression were assayed by real-time quantitative PCR and Western blot assays. The effects on cell invasiveness and migration were evaluated by wound healing and transwell assays. The antitumor activity of 10% YYGJ serum was compared to that of blank control, 10% rat serum control and 5-fluorouracil(FU). HepG2 and MHCC97H cell proliferation was inhibited by YYGJ serum in a time- and concentration-dependent manner. Cells accumulated in G0/G1 and apoptosis was increased in both cell lines by 10% YYGJ serum. The effects of apoptosis in 10% YYGJ serum were weaker than those in response to 5-FU. E-cadherin and Smad4 expression were upregulated by 10% YYGJ serum, but c-Myc, vimentin and MMP2 expression were downregulated in both hepatoma cell lines. The protein expression of Smad4 in HepG2, and mRNA expression of MMP2 and E-cadherin in both cell lines had no difference between 10% YYGJ serum and 5-FU treated groups. Cell invasion and migration were decreased by 10%YYGJ serum while cell cytotoxicity was shown in 5-FU treated group. YYGJ drug-containing serum inhibited HepG2 and MHCC97H cell proliferation, induced apoptosis, and regulated the expression of tumor-related genes and proteins. It reduced tumor cell invasion and migration. Further study to investigate the antitumor activity of YYGJ is warranted.