To investigate the role of integrin and G protein pathways in the mechanotransduction process within MCCs and explore the possible crosstalk between the two traditional signal pathways, in vitro-cultured rabbit MCCs were treated with pressure. The mRNA level of alpha5beta1 integrin was determined by in situ hybridization and the distributions of vinculin, Galphaq/11 protein, F-actin and intracellular calcium were studied with a laser scanning confocal microscope. Increased integrin alpha5beta1 expression, enhanced stress fiber assembly, elevated G protein and vinculin level and up-regulated IP(3) channel sensitivity were found in the mechanotransduction process of MCCs under pressure. Furthermore, the vinculin and the Galphaq/11 were observed co-localized with each other, and the F-actin reassembly and stress fibers formation could be inhibited by intracellular calcium channel blocking, which gave direct evidence that the traditional integrin-mediated or G protein-mediated signaling pathways coordinately regulate the function of MCCs under mechanical stimulation.