Cell cycle control in isoproterenol-induced murine salivary acinar cell proliferation.
The eukaryotic cell cycle is a summary of a complex network of signal transduction pathways resulting in both DNA replication and cell division. Cyclin-dependent kinases (CDKs) control the cell cycle in all eukaryotes, whereas other proteins, known as cyclins, act as their regulatory subunits. Chronic injection with isoproterenol (ISO) can induce acinar cell proliferation in rodent salivary glands. Cyclins and CDK proteins from control and ISO-treated murine parotid acinar cells were detected by using Western blotting techniques. By comparing the expression of these cell cycle regulatory kinases in the parotid acinar cell transition from a quiescent state to a hypertrophic state, we found rapid increases in the protein levels of all CDKs, cyclin D and proliferating cell nuclear antigen (PCNA). The highest protein levels for CDKs and cyclins appeared at about 72 hr of ISO stimulation and were coincident with the highest rate of increase in gland wet weight. After 72 hr, the increase of both cell cycle protein and gland wet weight began to subside. By using a co-immunoprecipitation method, the following cell cycle regulators (CDK-cyclin complexes) were detected, CDK4-cyclin D, CDK2-cyclin E, CDK2-cyclin A, and cdc2-cyclin B, along with an increase in kinase activity over control untreated animals. Additionally, we detected significant decreases in the newly isolated CDK inhibitor (CKI) p27kip but not Wee 1 kinase. The increased levels of CKI correlated with a decrease in kinase activity of CDK/cyclin complexes by 144 hr of chronic isoproterenol treatment. Our data suggest that the holoenzymes for cell cycle control (cyclin-CDK complexes) function as a final regulatory mechanism leading to salivary gland acinar cell proliferation. The gradual decline in protein levels of the CDKs and cyclins after 3 days of chronic treatment further indicates that ISO-induced proliferation of parotid acinar cells is self-limiting and non-tumorigenic.
Zeng
T
T
Department of Oral Biology, University of Florida, Gainesville 32601, USA.
Yamamoto
H
H
Bowen
E
E
Broverman
R L
RL
Nguyen
K H
KH
Humphreys-Beher
M G
MG
D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D018844:Cyclin-Dependent Kinases / Q000032:analysis*; D016213:Cyclins / Q000032:analysis*; D004261:DNA Replication; D005260:Female; D006657:Histones / Q000737:chemistry; D007545:Isoproterenol; D051379:Mice; D008807:Mice, Inbred BALB C; D009929:Organ Size / Q000187:drug effects; D010306:Parotid Gland / Q000166:cytology / Q000187:drug effects*; D010766:Phosphorylation; D011233:Precipitin Tests; D015398:Signal Transduction / Q000502:physiology
D016213:Cyclins; D006657:Histones; D018844:Cyclin-Dependent Kinases; D007545:Isoproterenol