The Anti-cancer Activity Of Triterpenoid From Submerged Fermentation Inototus Obliquus To Ovarian And Lung Cancer Cells

Inonotus obliquus is an important folk traditional medicine,showing antibacterial,antiviral,antifungal,liver preservation,antioxidation,anti-inflammatory and anti-tumor activity.Triterpenoid is an important medicinal ingredient in Inonotus obliquus.The purpose of this paper is to study The inhibitory effect of biological activity of triterpenoids from Inonotus Obliquus under submerged fermentation on the lung cancer cell line?A549,PC9?and ovarian cancer cell lines?ES2,SKOV3?.The MTT results showed that the crude triterpenoids had significant inhibitory activity on these four cell lines,and fraction 9 isolated by silica gel column chromatography showed better effects on ES2,SKOV3 and PC9 cell lines,while fraction 7 showed better effects on A549,PC9 and ES2 cell lines.The cell cycle experiments showed that the blocking action of the crude triterpenoids to ES2,SKOV3,A549 and PC9 occurred in G0/G1 phase.We analyzed the triterpenoid components of samples from the control culture medium by HPLC.Then we tried to optimize the medium of Inonotus obliquus on the triterpenoid production by adding the bark and shell of Eucommia ulmoides.The results of the study are as follows:1.After treatment of triterpenoids with different concentrations for 3 days,the IC₅₀ values of the crude triterpenoids to the ovarian cancer cell line ES2 was 21.04?g/mL,the IC₅₀ values to the cell strain SKOV3 was 33.84?g/mL,the IC₅₀ values of the lung cancer cell strain A549 was 28.46?g/mL,and 21.38?g/mL to the cell strain PC9.After treatment of triterpenoid with different concentrations for 6 days,the IC₅₀values of the crude triterpenoids to the ovarian cancer cell line ES2 was 17.60?g/mL,the IC₅₀ values to the cell strain SKOV3 was 10.94?g/mL,the IC50 values of the lung cancer cell strain A549 was 23.50?g/mL,and 19.67?g/mL to the cell strain PC9,there is a positive correlation with the time.2.the IC₅₀ values of fraction 9 on ES2,SKOV3 and PC9 cell lines were 22.67?45.31 and 28.46?g/mL,respectively,respectively,the IC₅₀ of fraction 7 on ES2,SKOV3 and PC9 cell lines were 31.91?31.01 and 29.09?g/mL,both of fraction 9and 7 showed better anti-cancer activities than the crude triterpenoids possessed in the same expriment.3.The cell cycles of PC9,A549 and ES2,SKOV3 cell lines were blocked in G0/G1 phase,and when treated with 16?g/mL triterpenoids,the proportion of cells in G1 phase of cell cyle of PC9?A549?ES2?SKOV3 increased by 32.7%?41.5%?5.7%?9.5%,respectively..4.Study on apoptosis indicated that triterpenoid can induce apoptosis of PC9,A549 and ES2,SKOV3 cell lines5.HPLC results showed that betulin may be one of the anticancer components,which may show better anticancer activity when it is in combination with inotodiol?trametenolic acid.6.After the addition of Eucommia ulmoides shell,the biomass of Inonotus Obliquus increased by 101.3%,the content of triterpene increased by 7.4%,and the crude triterpene products increased by 87.62%.After the addition of Eucommia ulmoides bark,the biomass of Inonotus Obliquus decreased by 2.14%,the content of triterpene increased by 109%,and the crude triterpene products increased by 80.21%.