8-bromocyclic-gmp and Cell-Transformation--Viral

A caprine luteal cell line (tsCLC-D) that synthesizes progesterone (P4) was established following by transformation with a temperature-sensitive A209 (tsA209) mutant of simian virus 40 (SV40). The transformed cells have temperature-sensitive for morphology, cell propagation and progesterone steroidogenesis. At the permissive temperature of 34 degrees C, these cells were spindle-shaped and grew with a similar rapidity as tumor cells. However, at the nonpermissive temperature of 40 degrees C, the cells have exhibited a round shape and ceased to proliferate because the gene for maintenance of transformation was not expressed. The tsCLC-D cell line responds to 8-Br-cyclic AMP, 22-hydroxycholesterol and pregnenolone treatment with an increase in progesterone biosynthesis. This cell line still express StAR protein, 3beta-HSD and P450scc enzyme of three kinds of steroidogenic protein and enzymes, this characteristic is similar to normal luteal cell. However, the addition of any doses oLH did not increase progesterone secretion. We speculate that tsCLC-D might lose the responsiveness to gonadotropins during the immortalization process, while retaining steroidogenic enzyme activity and progesterone production. To our knowledge, this is the first report of a stable cell line derived from corpus luteum of ruminant. The tsCLC-D retains steroidogenic capacity, which will make this cell line useful for the studies of regulation of steroidogenesis.

Topics: Animals; Cell Line, Transformed; Cell Separation; Cell Transformation, Viral; Cyclic GMP; Female; Goats; Hormones; Luteal Cells; Simian virus 40; Steroids; Temperature