Osteopontin increases hepatocellular carcinoma cell growth in a CD44 dependant manner

Figure 6 CD44 knockdown inhibits osteopontin-a-induced cell growth in Huh-7 and HeLa cells. A panel of 3 Stealth siRNA’s were investigated for their ability to limit CD44 mRNA expression using polymerase chain reaction (A), further validation was achieved by examining protein reduction of CD44 using Western blotting (B). (C) Huh-7 or (D) HeLa cells were seeded at a density of 7 × 10⁴ cells per well in a 12-well plate and a CD44 siRNA pool containing Stealth siRNA1 and 2 was transfected into the cultures 24 h prior to placing conditioned media from osteopontin (OPN)-A Huh-7 stable cell lines or Huh-7 cells alone onto the cells. Cell numbers were monitored daily using trypan blue exclusion. CM: Conditioned media; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase. ^aP < 0.05.