CD151 promotes cancer cell metastasis via integrins α3β1 and α6β1 in vitro.
MOLECULAR MEDICINE REPORTS(2012)
Abstract
CD151 is a member of the tetraspanin family that is implicated as a promoter of the tumor metastasis of malignant cells. Tetraspanins form membrane complexes with integrins. In the present study, we constructed a CD151-AAA mutant to assess the roles of Rac, cdc42 and phospho-Rac/cdc42 (P-Rac/cdc42) and the effects of CD151-integrin complexes on the proliferation, migration and invasion of HepG2 cells. The pAAV-CD151 and pAAV-CD151-AAA mutant plasmids were constructed and used to transiently transfect HepG2 cells using the Qiagen Attractene transfection reagent. Following transfection, the expression of CD151 was determined by western blotting. A cell proliferation assay was performed using the cell counting kit-8 (CCK-8) method, cell migration was assessed by a cell wound-healing assay and cell invasion was evaluated in microchemotaxis chambers using FBS as the chemotactic stimulus. The potential involvement of various signaling pathways was explored using relevant antibodies. The association between CD151 and integrins was evaluated by immunoblotting analysis. We found that CD151 promoted cell proliferation, migration and chemotaxis and increased P-Rac/cdc42 activity. The CDI51-AAA mutant had reduced cellular proliferation, migration and invasion compared with the CD151 mutant. Moreover, the CD151-AAA mutant abrogated the association between CD151 and integrins. These data suggest that CD151 forms complexes by interacting with integrins, particularly alpha 3 beta 1 and alpha 6 beta 1, and thereby affects the functioning of the HepG2 cells. The mechanism is possibly related to the Rac,cdc42 and P-Rac/cdc42 signaling pathways.
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Key words
tetraspanin,CD151,CD151-AAA,hepatocellular carcinoma,metastasis
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