Pilot Study On Antitumor Substances Produced By The Marine Fungus Penicillium Glaucum HUC-5

The research of marine natural products has been a hot field in china and abroad. Screening bioactive marine natural products is another important work in natural products research. We screened marine microbes producing antitumor material from the26marine fungi stored in our lab by MTT assay. The results showed that fermental broth from nine of the twenty-six strains had antitumor activity, six strains showed strong cytotoxic activity against HeLa, five strains against PC-12among those nine microbes. Two of the nine strains had both anti-HeLa and PC-12cell lines activity. The antitumor activities of acetic ether extracts from marine fungus HUC-5broth were studied.The HUC-5was identified as Penicillium glaucum according to its colony morphological characteristics and conidial fructificationSome properties of the HUC-5extract were studied. The result showed that antitumor activity of the extract was well maintained at40-50℃compared to room temperature25℃While its inhibit rate started to decrease at more than60℃, the activity decreased about66%at80℃, and only22.6%activity was preserved. The antitumor substances could dissolved in organic reagent such as ethyl acetate, methanol et al, the best reagent was ethyl acetateThe effects of HUC-5extract on inhibiting tumor cells were studied. The results showed that HUC-5extract had broad-spectrum anti-tumor activity. It could be against HeLa, PC-12, BT474, B-16, Hep-2cell lines in vitro. The inhibition rate to Hep-2was the highest among those cells. The HUC-5extract inhibited proliferation of Hep-2cells in a concentration-dependent and time-dependent manner, and its IC50value was0.12mg/mL. Hep-2cells treated by HUC-5extract appeared chromatin condensation, cell shrinkage and theformation of apoptotic bodies. Comet assay result suggested that Hep-2cells were severely damaged by marine fungus HUC-5extract. Agarose gel electrophoresis showed obvious DNA ladder on the gel. Results of flow cytometry assay indicated that Hep-2appeared obvious apoptosis at16h and32h, whose apoptosis rate was45.85%and86%respectively. The HUC-5extract could inhibit proliferation of Hep-2cells by inducing apoptosis.Five components were isolated by Sephadex LH-20from HUC-2extract with15.65%10.69%14.48%16.99%18.11%activity restectively.