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. Author manuscript; available in PMC: 2010 May 1.

Published in final edited form as: Stem Cells. 2009 May;27(5):1006–1020. doi: 10.1002/stem.30

CD44 knockdown by lentivirus-mediated shRNA and the significant reduction of the tumorigenic ability of gastric cancer stem cells. (A): Fluorescence-activated cell sorter analysis of CD44 knockdown in MKN-45 cells by lentivirus-mediated human CD44-specific shRNA. Left: scramble shRNA as control; right: human CD44-specific shRNA. CD44 expression in MKN-45 cells was successfully knocked down. CD44 expression in MKN-74 cells was also downregulated to a similar level. (B): Spheroid colony formation of CD44 downregulated MKN-45 and MKN-74 cells. After 4 weeks of culture, CD44 shRNA-infected cells produced significantly lower number of colonies than scramble shRNA virus-infected cells. (C): Skin tumors produced by CD44 downregulated MKN-45 cells. CD44 shRNA-infected MKN-45 cells as well as scramble shRNA virus-infected cells were injected under the skin of SCID mice, respectively, and after 4 weeks, although all mice had tumors, CD44 shRNA virus-infected cells produced much smaller tumors than scramble shRNA virus-infected cells. CD44 downregulated MKN-74 cells also showed similar results. (D): Spheroid colony formation of high CD44-expressing and low CD44-expressing populations in MKN-45 and MKN-74 cells. After 4 weeks of culture, high CD44-expressing cells produced significantly larger number of colonies than low CD44-expressing cells. (E): Skin tumors produced by high CD44-expressing MKN-45 cells. High CD44-expressing as well as low CD44-expressing populations in MKN-45 cells were injected under the skin of SCID mice, respectively, and after 4 weeks, high CD44-expressing cells produced much larger tumors than low CD44-expressing cells. High CD44-expressing MKN-74 cells also showed similar results. Abbreviations: FITC, fluorescein isothiocyanate; PE, phycoerythrin; shRNA, short hairpin RNA.

CD44 knockdown by lentivirus-mediated shRNA and the significant reduction of the tumorigenic ability of gastric cancer stem cells. (A): Fluorescence-activated cell sorter analysis of CD44 knockdown in MKN-45 cells by lentivirus-mediated human CD44-specific shRNA. Left: scramble shRNA as control; right: human CD44-specific shRNA. CD44 expression in MKN-45 cells was successfully knocked down. CD44 expression in MKN-74 cells was also downregulated to a similar level. (B): Spheroid colony formation of CD44 downregulated MKN-45 and MKN-74 cells. After 4 weeks of culture, CD44 shRNA-infected cells produced significantly lower number of colonies than scramble shRNA virus-infected cells. (C): Skin tumors produced by CD44 downregulated MKN-45 cells. CD44 shRNA-infected MKN-45 cells as well as scramble shRNA virus-infected cells were injected under the skin of SCID mice, respectively, and after 4 weeks, although all mice had tumors, CD44 shRNA virus-infected cells produced much smaller tumors than scramble shRNA virus-infected cells. CD44 downregulated MKN-74 cells also showed similar results. (D): Spheroid colony formation of high CD44-expressing and low CD44-expressing populations in MKN-45 and MKN-74 cells. After 4 weeks of culture, high CD44-expressing cells produced significantly larger number of colonies than low CD44-expressing cells. (E): Skin tumors produced by high CD44-expressing MKN-45 cells. High CD44-expressing as well as low CD44-expressing populations in MKN-45 cells were injected under the skin of SCID mice, respectively, and after 4 weeks, high CD44-expressing cells produced much larger tumors than low CD44-expressing cells. High CD44-expressing MKN-74 cells also showed similar results. Abbreviations: FITC, fluorescein isothiocyanate; PE, phycoerythrin; shRNA, short hairpin RNA.