The Study Of The Anti-tumor Effect And Mechanism After Oral Administration Of The Proteins(LFP_91-3) From The Fermentation Liquor Of Lentinus Edodes C_91-3

Objective:Cancer is a major threat to human health.Lentinus edodes (Berk) Sing,also named as champignon, belongs to Eumycophyta, basidion-ycetes sub-division, hymenomycoid range,agaric order,tricholomoid genus. Lentinus is one of the favorite edible and medicinal fungi in the world,which contains a variety of effective medicinal ingredients. Studies have found that Lentinus can improve the body's immunity, prevent cancer cells multiplicating. Currently lentinan as an important biological response modifier, has been widely applied in cancer immunotherapy. In addition to polysaccharides, Lentinus also contains a large number of proteins, amino acids and trace elements. The enhancement or potentiation of host defense mechanisms has been recognized as a possible means of inhibiting tumor growth without harming the host. Studies on the biological activities and the antitumor mechanisim of the proteins from Lentinus edodes has been reported rarely.This seminar had studied the Lentinus edodes for many years and finally got strain C91-3 which has obviously anti-tumor and anti-bacterium functions after fermentation. The components of fermentation liquor and fresh body are basically similar. Besides polysaccharides and amine acids, the fermentation liquor contains kinds of protein components. Especially the protein component of its fermentation liquor (LFP91-3) can kill tumor cells directly. The experiment showed that LFP91-3C has significant anti-tumor effect both in vitro and in vivo. Based on previous experiments, The immuno-mechanism of oral liquid(LFP91-3) from the fermentation liquor of Lentinus edodes C91-3 was studied in this experiment.Methods:After fermentation, centrifugation, filtration, salt precipitation, dialysis, extraction and concentration with vacuum dryer, do the following experiments.(1) Effect of oral LFP91-3 liquid on life span in mice bearing S180 tumors60 male and female KunMing mice bearing S180 tumors were divided into 6 groups equally and randomly at 10 mice per group:distilled water control group, CTX (ctxclophosphamide) treatment group, LNT(Lentinan) treatment group, LFP91-3Ⅰ,ⅡandⅢtreatment groups.24 hours after implanted with tumor, distilled water control group received an intragastric dose of distilled water 0.5ml per mouse every another day for 8 days. At the same time, CTX group, LNT group, LFP91.3Ⅰ,ⅡandⅢgroups received one intragastric dose of CTX 3mg, LNT 0.5mg, LFP91-3250μg,500μg,1000μg. The therapeutic effect was determined by life span, which discuss the feasible dose.(2) Effect of oral LFP91-3 liquid on spleen lymphoctxte multiplying rate within mice bearing tumor20 male and female KunMing mice bearing S180 tumors were divided into 2 groups equally and randomly at 10 mice per group:distilled water control group, LFP91-3 treatment groups. Distilled water 0.5ml per mouse every another day for 8 days, LFP91-3500μg respectively in the same way. All mice were killed and excised the eyeballs to draw blood separately on the 16th day after bearing tumor. Spleen lymphoctxte multiplying rates:got the cell of mice spleen to concentration 5×106/ml. The experimental groups consist of the cell of mice spleen in the same concentration (per hole 200μl) and ConA liquid (50μg/ml) 20μl. The spleen cell control group, spleen cell and RPMI-1640 liquid.They were cultivated in incubator for 68 hours. Then the liquid was centrifuged and dumped. All the holes were added by MTT liquid 10μl and continued cultivating for 4 hours then added DMSO liquid 100μl to measure OD by enzyme sign instrument.(3)Effect of oral LFP91-3 liquid on serum concentration of IL-2 and TNF-αwith mice bearing tumorBlood serum was got from the mice eyeballs in the same way mentioned above. Use the Kit to get the serum concentration of IL-2 and TNF-αwithin mice bearing tumor.Results:1. Oral LFP91.3 liquid could prolong the life span of mice bearing S180 tumor, whose life span was 24.80d,25.10d,25.60d longer than that of distilled water group(17.80d) (P<0.05),which is the similar as that of CTX group (24.70d) and LNT group (24.30d) (P>0.05).2. Oral LFP91-3 liquid has higher inhibition rate than distilled water group (P<0.05) within mice bearing tumor. The spleen lymphoctxte multiplying rate of LFP91-3 groups was 0.44±0.14,which was higher than that of distilled water group 0.18±0.09.3. Oral LFP91.3 liquid could promote serum concentration of IL-2 and TNF-αwith mice bearing tumor. Serum concentration of IL-2 and TNF-αof LFP91-3 groups was 35.05±17.29 pg/ml,84.27±16.88 pg/ml, which was higher than that of distilled water group 30.5±3.58 pg/ml,48.33±16.70 pg/ml (P<0 .01).Conclusion:Oral LFP91.3 liquid has significant anti-tumor effect in vivo. Oral LFP91.3 liquid could prolong the life span of mice bearing tumors and inhibit solid tumor cells, which has the same role as CTX and LNT. Oral LFP91-3 liquid could also enhance immunity system to kill cancer cell.This research may provide new theory bases for developing anti-tumor drugs. It is believed in the near future, the LFP91.3C certainly can benefit the human.