ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of serum containing Chinese medicine (CM) Sanpi Pingwei (, SPPW) formula on the proliferation and apoptosis of human SGC-7901 cells and the possible mechanism.</p><p><b>METHODS</b>Serum containing CM SPPW formula (SPPW serum) was prepared by a serum pharmacology method. Human SGC-7901 cells were incubated with SPPW serum at three different concentrations and with the anticancer drug 5-fluorouracil (5-FU), respectively. Cell proliferation was assessed by MTT assay, and cell apoptosis was detected by flow cytometry assay. Real-time quantitative polymerase chain reaction (RT-PCR) and Western blot assay were employed to confirm the expressions of Bcl-2, Bax and p53 in SGC-7901 cells at mRNA and protein levels, respectively.</p><p><b>RESULTS</b>SPPW serum suppressed the proliferation of SGC-7901 cells in a time- and dose-dependent manner. The colony forming rate of negative control was 48.2%, while those in the three SPPW serum groups and the 5-FU group decreased significantly (P<0.01). The number of colony forming units in the SPPW high dosage group was significantly smaller than that in the 5-FU group (P<0.01). MTT assay showed that SPPW serum restrained the proliferation of SGC-7901 cells, and the inhibition rate increased significantly in a dose-dependent manner. Annexin V/PI Assay suggested that SPPW serum induced the apoptosis of SGC-7901 cells significantly. RT-PCR and western blot assay indicated that SPPW serum upregulated the protein and mRNA expression levels of Bax and p53 in SGC-7901 cells, but downregulated the protein and mRNA expressions of Bcl-2.</p><p><b>CONCLUSIONS</b>SPPW formula inhibits the proliferation of SGC-7901 cells in vitro and induces the cell apoptosis. It plays an anticancer role by regulating the expressions of Bax, p53 and Bcl-2 in SGC-7901 cells.</p>
Subject(s)
Animals , Humans , Rats , Apoptosis , Blotting, Western , Cell Line, Tumor , Cell Proliferation , Drugs, Chinese Herbal , Pharmacology , Fluorouracil , Pharmacology , Gene Expression Regulation, Neoplastic , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Serum , Chemistry , Tumor Stem Cell Assay , Tumor Suppressor Protein p53 , Genetics , Metabolism , bcl-2-Associated X Protein , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To clarify the action and possible mechanisms of SPPW, a Chinese herbal preparation consisting of Herba Scutellariae Barbatae, Radix Astragalus, Radix Glycyrrhizae, etc., in suppressing the metastasis of human gastric cancer, by way of observing its effect on the invasive and metastatic capacities of gastric cancer cells.</p><p><b>METHODS</b>In vitro serial sub-cultured human gastric cancer cell line SGC-7901 at the logarithmic growth phase were randomly divided into 5 groups, i.e. the negative control group, the 4 treatment groups intervened respectively with SPPW at three different doses (high, middle, and low), and 5-FU. The adhesion capacities of gastric cancer cells to matrigel were detected by MTT assay 48 h after intervention. The invasive and migratory capacities of gastric cancer cells were determined by Transwell assay. The mRNA and protein expressions of metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF) in gastric cancer cells were detected by Western blot and Real-time polymerase chain reaction (RT-PCR) respectively.</p><p><b>RESULTS</b>Compared with the negative control group, the adhesive, invasive and migratory capacities of gastric cancer cells were all significantly inhibited in the four treatment groups (P<0.05, P<0.01). In addition, the protein and mRNA expressions of MMP-9 and VEGF were down-regulated (P<0.01). Significant dose-dependent relation existed in the three SPPW treatment groups (P<0.01). Compared with the 5-FU treatment group, the high dose SPPW treatment group showed significant difference in inhibiting the adhesive and metastatic capacities of gastric cancer cells, lowering VEGF protein expression, and mRNA expressions of MMP-9 and VEGF (P<0.01).</p><p><b>CONCLUSIONS</b>SPPW could lower the adhesion of gastric cancer cells to matrigel, and lower the invasion and migration of gastric cancer cells. Meanwhile, it could down-regulate the mRNA and protein expressions of MMP-9 and VEGF, which may possibly be one of its mechanisms for influencing the invasion and metastasis of gastric cancer cells.</p>