cytochrome-c-t and resazurin

To investigate the inhibition effect of Ganoderma applanatum extract (GAEAE) on human gastric cancer cell lines and apoptosis mechanism, Alamar Blue assay was used to assess the inhibition and apoptosis-inducing effect of the GAEAE on proliferation of SGC-7901 cells, the DNA ladders of the apoptosis cells was done, the mRNA expression of p53, Bax, Bcl-2 and JNK were analysed by reverse transcription-polymerase chain reaction. The levels of the Cyt C and the p53, Bax/Bcl-2, JNK proteins and the caspase-3 activity in the SGC-7901 cells were measured with ELISA kits. Our data showed that the GAEAE markedly inhibited the proliferation of SGC-7901 cells in a dose-dependent manner. The expression of Bcl-2 protein was down-regulated and Bax, c-jun, p53 protein expressions were up-regulated by the GAEAE treatment in SGC-7901 cells. The activity of caspase-3 was markedly increased and the Cty C was markedly released into cytoplasm from the mitochondria after GAEAE action. In conclusion, our results indicated that the GAEAE could enhance the sensitivity of SGC-7901 cells to the c-jun, p53, Bax and Bcl-2 induced apoptosis and provided a promising approach to anti-human gastric cancer therapy with Ganoderma applanatum.

Topics: Apoptosis; bcl-2-Associated X Protein; Caspase 3; Cell Line, Tumor; Cell Proliferation; Cytochromes c; DNA Fragmentation; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Ganoderma; Gene Expression Regulation, Neoplastic; Humans; MAP Kinase Kinase 4; Mitochondria; Oxazines; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-jun; Reishi; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stomach Neoplasms; Tumor Suppressor Protein p53; Xanthenes