KB 사람 구강 편평 암세포에서 Lyophyllum shimeji 로부터 분리한 단백질 추출물의 항암활성

Abstract

There are various mushroom proteins with interesting biological activities, such as flavonoid, lectins, ribosome inactivating proteins and other proteins, which have become popular sources of natural anti-tumor agents. Many mushrooms have been reported to have cancer-preventing properties. However, little research has been conducted verifying the anti-tumor activities of Lyophyllum shiemeji mushrooms. This study examined the effect of protein extract fromLyophyllum shiemeji on anti-cancer effect and molecular mechanism of KB human oral squamous cell carcinoma. To understand of molecular mechanism whether its cell death is related with apoptosis pathway, performed DNA fragmentation, western blot and LIVE-DEAD cell staining. to explore the anti-cancer activity of the protein extract from Lyophyluum shimeji, we performed MTT assay. KB cells were treated with protein extract from Lyophyllum shiemeji at 100 ㎍/㎖, 125 ㎍/㎖, 150 ㎍/㎖, 175 ㎍/㎖ and 200 ㎍/㎖ for 24 hours. The protein extract from Lyophyllum shiemeji displayed the inhibitory activity against KB cells with an IC50 value of 150 ㎍/㎖ and the strongest inhibition percent was 91.66% at the concentration of 200 ㎍/㎖. It was observed that the protein extract from Lyophyllum shiemeji induced genomic DNA ladder formation. In addition, it was confirmed that protein extract fromLyophyllum shiemeji induced significantly increase cleaved caspase-7 activation and PARP cleavage in KB cells. Activation of cleaved caspase-9 induced by protein extract from Lyophyllum shiemeji suggested that protein extract from Lyophyllum shiemeji -induced signaling was mediated through a mitochondrial related pathway. Furthermore, protein extract from Lyophyllum shiemeji treatment KB cells dramatically increase the red color (apoptotic cell) by fluorescence microscopy. These findings demonstrated that protein extract fromLyophyllum shiemeji -induced apoptosis in KB human oral squamous cell carcinoma involves the mitochondrial intrinsic apoptotic signaling pathway. It also suggest that protein extract from Lyophyllum shiemeji be developed as a chemotherapeutic agent and adjvant for the management of oral squamous cell carcinoma.