Polysaccharides Of Ganoderma Lucidum Selenium And Its Anti-tumor, And Induce Tumor Cell Apoptosis

Both polyssacharide and selenium have the function of prevention and treatment of many diseases. The researches on the activities of polyssacharide and selenium have been performed deeply, but it still stays on the primary stage in the world to combine the polyssacharide and selenium into selenium containing polyssacharide organically. Selenium containing polyssacharide in Ganoderma Lucidum (SeGLP) is a kind of half natural and half synthesized polyssacharide of Fungi enriched selenium, whose structure differs from other ordinary polyssacharides —— it contains a special bond between selenium atom and oxygen atom. SeGLP has many physiological functions, including that it can resist heavy mantal poisoning and active oxygen injury, can influence cell cycle and inhibit the synthesis of DNA of the cancer cell, and induce the apoptosis process of tumor cell.Selenium containing mycelium of Ganoderma Lucidum could be obtained by biotransformation. The mycelium was exstracted with hot water and alkali, and raw selenium containing polyssacharide (SeGLP) was obtained. Then the raw SeGLP was handled with protein and pigment removal and precipitation, and was purified and isolated by DEAE-cellulose column chromatography, the sub-fractions of raw SeGLP were obtained. Hca-f cells and S180 cells were inoculated hypodermically at the concentration of 1×106 cells per mouse and the SeGLP were resolved into aseptically distilled water to make injection liquid. After continuous 15 days injection, then calculating the inhibition rate of tumor, testing the activities of GPx and SOD and the content of MDA both in serum and liver in mice in order to observe the relationship between antioxidation and antitumor of SeGLP.The induction of tumor cell apoptosis of SeGLP could be detected by morphological observation, DNA ladder technology, MTT colorimetric estimation, Flow cytometer detection, immunity histochemistry and PCR-SSCP technology. The results showed that Ganoderma Lucidum could grow normally in the improved PDA culture added with Na2SeO3 at the concentration of 400μg/g, and the harvest rate of mycelium was about 1%. The harvest rate of SeGLP more than 30%. SeGLP-1A,SeGLP-1B,SeGLP-2A,SeGLP-2B,SeGLP-3A and SeGLP-3B six SeGLPs could be isolated and purified, and the content of selenium of seriese SeGLPs were different. SeGLP had strong inhibition effect on Hca-f cells and S180 cells, and the inhibition rate could reach 32-60% and 42-67% respectively. SeGLP could enhance the activities of GPx and SOD and decrease the content of MDA both in serum and liver in mice, which indicated that SeGLP had obviously antioxidation function, and it could inhibit the growth of tumor cell by enhancing the antioxidant ability in body.SeGLP could induce the apoptosis of K562 cells significantly, cell membrane was damagede, cell transmuted or produced gemmae, constricted region and apoptotic bodies, and the transparency and brightness of cells were declined. The proliferation function and the number of active cells were both decreased. Cell crenation, condensation and margination of nuclear chromatin by cytoplasimic hypervacuolization could be seen by morphological ovservation. Typical DNA ladder could be seen by agar-gel electrophoresis. The results of flow cytometer show that significant apoptotic peak and SeGLP can stop the cell cycle at the sub-G0/G1 stage and can reduce the mutant rate of the P53 gene. The results of PCR-SSPC show that SeGLP can lead the mutation of the sixth extron of the P53 gene, thus induce the apoptosis procedure of tumor cells.