In order to find out the regulatory function of hormone release of malignant choriocarcinoma, we cultured BeWo cell lines in vitro. The control was cultured in cAMP and insulin free medium and for the experimental group, we add 0.1, 0.5 and 1.0mM
of
cAMP〈Group 1〉, 1, 10 and 20μM of insulin 〈GroupⅡ〉and, 0.1 mM and 1 μM, 0.5mM and 10 1.0mM and 20μM of cAMP nd insulin〈GroupⅢ〉, respectively to the medium, then cultured for 3 days in vitro. Release of β-hCG, estradiol and progesterone
was
measured by radioimmunoassay.
In order to find out the effect of cAMP and insulin on cell proliferation, we measured the number of cells by using hemocytometer after 3-day culture.
@ES The results were as follow.
@EN 1. β-hCG release was increased in control and all experimental groups as progressing the days of culture. Release of β-hCG was significantly increased in proportion to the amount of cAMP and decreased in proportion to the amount of insulin
as
compared with those of control in all groups. In GroupⅢ, inhibitory effects of insulin was more predominant than cAMP of low concentration, and no inhibitory effect of insulin was found in the cases of stimulatory effect of high concentration of
cAMP
in spite of high concentraion of insulin.
2. Estradiol release was relatively increased in control and Group Ⅱ and GoupⅢ, but decreased in Group I with low concentration of cAMP as progressing the days of culture. In GroupⅠandⅡ, release of estradiol was decreased as compared with
that
of
control. In GroupⅢ, there was no statistical significance as compared with control.
3. Progesterone release was relatively increased in control and experimental groups as Progressing the days of culture, In Group I(except low concentration group), release of progesterone was increased on the 1st day of culture as compared with
that of
control. Howevere, in high cAMP concentration group, release was decreased on the 2nd day of culture as compared with control. In Group Ⅲ, there was a tendency toward reduction of progesterone release as compared with control.
4. Number of cells was significantly increased after 3-day culture. In Group I with high concentration of cAMP, number of cells was decreased, which suggested inhibition of cell proliferation, In Group Ⅱ with high concetration of insulin, number
of
cells was incerased, which suggested stimulatory effect of insulin on cell proliferation. In Group Ⅲ, neither stimulatory nor inhibitory effect was found.
From the above results, it is thought that cAMP and insulin are related to synthesis and release of hCG and estradiol in BeWo cell line which is one of malignant trophoblastic cell and are insignificantly related to release of progesterone as
compared
withβ-hCG or estradiol. It was also found that cAMP inhibited and insulin stimulated BeWo cell-proliferation.
|