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. Author manuscript; available in PMC: 2008 Sep 15.
Published in final edited form as: Hepatology. 2008 Apr;47(4):1211–1222. doi: 10.1002/hep.22202

Fig. 3. Knockdown of SULF2 decreases HCC cell proliferation and migration.

Fig. 3

(A) Plasmid constructs expressing an shRNA targeting the SULF2 mRNA or the empty vector plasmid pSS-H1p were stably transfected into Huh7 cells, which expressed high levels of SULF2 mRNA. SULF2 mRNA levels were quantitated by real-time RT-PCR in Huh7 Vector (Huh7 Vec) and the two SULF2 shRNA clones, designated Huh7 SULF2shRNA-3 (Huh7 sh-3) and Huh7 SULF2shRNA-4 (Huh7 sh-4). SULF2 mRNA levels were profoundly suppressed by the shRNA plasmid. (B) Huh7 Vec, Huh7 sh-3, and Huh7 sh-4 cells were plated into 6-well plates at 1 × 105 cells per well for the indicated time periods, and the total number of viable cells from each well was counted after trypan blue staining (P < 0.05 at days 4 and 5). (C) Huh7 Vec, Huh7 sh-3, and Huh7 sh-4 cells were plated into 96-well plates at 3000 cells per well. After 24 hours, cell proliferation was evaluated by the BrdU incorporation assay. Knockdown of SULF2 mRNA significantly decreased BrdU incorporation of both shRNA clones (P < 0.05). (D,E) Scratch wounds were made in confluent cell culture monolayers with a 20-µL pipette tip; photomicrographs of the wounds were taken at 0 and 24 hours thereafter. Relative migration of the wound edge was quantitated and showed a significantly decreased rate of migration in Huh7 sh-3 and Huh7 sh-4 cells when compared to Huh7 Vec cells (P < 0.05).