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Am J Physiol Cell Physiol 280: C1440-C1448, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 6, C1440-C1448, June 2001

Collagen gel overlay induces two phases of apoptosis in MDCK cells

Yang-Kao Wang1,*, Hsi-Hui Lin2,*, and Ming-Jer Tang1

1 Department of Physiology, National Cheng-Kung University Medical College, and 2 Department of Nursing, Chang-Jung Christian University, Tainan, Taiwan 70101, Republic of China

We previously demonstrated that collagen gel overlay induced cell remodeling to form lumen and apoptosis in Madin-Darby canine kidney cells. In the present study, we established that collagen gel overlay-induced apoptosis was initiated at areas exclusive of cell remodeling within 24 h (first phase) and extended into areas of cell remodeling within 48 h (second phase). Collagen gel overlay-induced apoptosis was accompanied by selective proteolysis of focal adhesion kinase (FAK), talin, p130cas, and c-src. Upon collagen gel overlay, FAK was initially degraded into a 90-kDa product during the first phase and subsequently into a 80-kDa product during the second phase. Collagen gel overlay-induced apoptosis of focal adhesion complex proteins and apoptosis of the first phase could be blocked only by a protease inhibitor cocktail. In addition, we found that both DEVD-fmk and ZVAD-fmk inhibited secondary proteolysis of FAK, but only ZVAD-fmk blocked collagen gel overlay-induced apoptosis of the second phase. Finally, collagen gel overlay-induced apoptosis and proteolysis of focal adhesion complex proteins were completely inhibited by the combination of protease inhibitor cocktail and ZVAD-fmk. Taken together, collagen gel overlay induces two phases of apoptosis; the first phase is dependent on proteolysis of focal adhesion complex proteins, and the second phase on activation of caspases.

Madin-Darby canine kidney cells; proteolysis; focal adhesion kinase

* Y.-K. Wang and H.-H. Lin contributed equally to this work.




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