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Soichiro Nakamura,Masahiro Ogawa,Hao Jing,David D. Kitts,Shuryo Nakai 한국식품영양과학회 2003 Journal of medicinal food Vol.6 No.4
Caco-2 and HCT-116 cells were used to access growth-inhibition and anti-invasion activity of recombinantcystatin C expressed in Pichia pastorisX33, G12W/H86V. The mutant G12W/H86V prepared by a pilot plant productionsystem showed more than 10% growth inhibition of Caco-2 cells at 0.56 56 n M concentrations. Growth-inhibited cells hadlower cathepsin L activity than the control cells that were not treated with the inhibitor. Conversely, the cathepsin B activitywas not changed by treatment with G12W/H86V. The in vitroanti-invasion test using HCT-116 cells showed that G12W/H86Vsuppressed the cell invasion by 15%, while its wild-type cystatin, aspartic protease inhibitor pepstatin A, and matrix metal-loproteinase (MMP) inhibitor MMP-2/MMP-9 inhibitor III did not supress cell invasion. These results indicate that the re-combinant cystatin C with higher protease inhibitory activity effectively retards the growth and invasiveness of human coloncarcinoma cells.