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Hybridoma monoclonal antibody 4F11 recognizes CD90+ liver cancer stem cells and inhibits their invasion
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Abstract:
Objective:To screen the monoclonal antibody (McAb) recognizing liver cancer stem cells and study its anticancer effects in vitro so as to provide antibody drug candidates for liver cancer stem cell-targeted therapy. Methods: Serum-free suspension culture and PKH26 staining was used to determine whether there were liver cancer stem cells in human hepatoma cell line MHCC97H. Expressions of 7 cancer stem cell biomarkers in MHCC97H cells and MHCC97H sphere cells, and co-expressions of antigens recognized by CD90 and different McAbs (3G7, 4F11, 11C9, 15B7, 15D2) in MHCC97H cells were examined by flow cytometry assay. The effects of McAb 4F11 on self-renewal and proliferation of MHCC97H cells and MHCC97H sphere cells were identified by serum-free suspension culture and CCK-8 assay. The effects of McAb 4F11 on in vitro invasion and migration of MHCC97 cells was examined by Transwell assay. Results: PKH26 staining results demonstrated that each MHCC97H sphere cell was proliferated and differentiated from one single liver cancer stem cell. The positive rate of CD90 in MHCC97H sphere cells were significantly higher than that in parent MHCC97H cells（\[18.0?7.5\]% vs \[2.3?1.0\]%，P<0.05). McAbs 3G7, 4F11, 11C9, 15B7 and 15D2 all CD90+MHCC97H cells in MHCC97H cells with the recognition rate of McAb 4F11 in CD90+MHCC97H cells was (47.2?44)%. The inhibitory rate of McAb 4F11 on MHCC97H sphere cells was significantly higher than that on MHCC97H cells（\[29.4?3.8\]% vs \[12.0?2.2\]%，P<0.05), and McAb 4F11 extremely suppressed the sphere formation ability of MHCC97H cells and inhibitory rate was (58.0?20.8)%. McAb 4F11 also significantly inhibited invasion and migration of MHCC97H cells in vitro (inhibitory rate: \[48.6?5.1\]% and \[47.6?3.6\]%). Conclusion: Hybridoma McAb 4F11 can specifically recognize CD90+ liver cancer stem cells, and inhibit invasion and migration of liver cancer cells. McAb 4F11 could be a antibody drug candidate for liver cancer stem cell targeted therapy.
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Project Supported:
Project supported by the National Science and Technology Major Project of China (No. 2011ZX09102-010-02, No. 2011ZX09401-030-4), and the National Key Basic Research Development Program (973 Program) of China (No. 2009CB521804)
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