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The methanolic extract from the branch of Lawsonia inermis was found to inhibit the invasion of HT1080 cells strongly. The extract and its EtOAc- and 1-BuOH-soluble fractions strongly inhibited the enzyme activities and the secretions of MMPs from HT1080 cells. We, therefore, tried to examine and estimate the active constituents among the constituents isolated. However, there are no active compound among the isolated compounds. Next, we tried to identify the selective binding protein of whole constituents of their extracts. As a result, ribosomal protein S18 (RPs S18, human), 60S ribosomal protein L18 isoform 1 (60S RPs L18 i1, human) and eukaryotic elongation factor 2 (eEF2, human) were detected as their selective binding and potential target proteins. To confirm that the proteins are target proteins or not, we examined the effects of their proteins on synthesis of MMPs using siRNA method. With the treatment of siRNA for RPs S18, 60S RPs L18 i1 and eEF2 proteins, the MMP-2 protein synthesis were downregulated in HT1080 cells. MMP-9 productions were also downregulated with the treatment of siRNA for RPs S18 and eEF2. From the results, we reached a conclusion that the extract was considered to have the potential to inhibit the production of MMPs proteins via inhibition of RPs S18 and eEF2 proteins, leading to inhibit the invasion in HT1080 cells.

Anti-invasive Activity of Lawsonia inermis Branch and Its Potential Target Protein