The Effect Of GnRH-a On Human Endometrial Carcinoma HEC-1B Cells And PTEN/C-myc Genes

ObjectiveRecently, epidemiological studies have found that the application of gonadotropin releasing hormone analogues(GnRH-a)is related to the degression of morbidity and mortality of many hormone-dependent tumors. But most studies have focused on the aspect of endometriosis, uterinemyoma, ovarian carcinoma,breast carcinoma and prostate,carcinoma etc .Compared with few about endometrial cancer.In this study,we observe the proliferative activity of different concentration triptorelin acetate and the expression of C-myc/PTEN mRNAfor the target of human endometrial carcinoma cell line HEC-1B.So,The result may be helpful to cure early endometrial cancer.Materials and Methods1. Materials:human endometrial carcinoma cell line HEC-1B;triptorelin acetate2. Methods:adenocarcinoma endometrium cell HEC-1B is cultured in the culture solution with triptorelin acetate of different concentration(0mol/L,10-9mol/L,10-8mol/L,10-7mol/L,10-6mol/L,10-5mol/L) at 24hours,48hours,72hours,96hours to test.Main observation1. At the same time,to detect the different group inhibition ratio with different concentration of triptorelin acetate;at the same concentration of triptorelin acetate, to detect the different group inhibition ratio at different time.Using MTT.2. To detect the expression of C-myc protein and PTEN protein in HEC-1B; Using immunohistochemistry ABC.3. Observe the morphology changes of HEC-1-B cells .Using inverted phase contrast microscope.4. To detect the expression of C-myc mRNA and PTEN mRNA in HEC-1B cells in different concentration of drug at the same time point.Using RT-PCR.Results1. As the concentration of drug goes high, the inhibition ratio raises up at the same time point, there is significantly difference among groups(P<0.05);as time lasts ,the inhibition ratio raises up at the same concentration of triptorelin acetate,there is significantly difference among groups(P<0.05).2. The expression of C-myc protein exists in HEC-1B cells and the PTEN protein is contrary.3. After the impact of triptorelin acetate,the cell density steps down obviously,cell becomes small,cell space becomes larger and vacuolus were noticed in cells;4. As the concentration of triptorelin acetate goes high,the expression of C-myc mRNA becomes lower and the PTEN mRNA becomes higher,there was a negative correlation between the two at the same point.Conclusion1. Triptorelin acetate can inhibit cell multiplication.This effect is time and dose dependent.It proves that the effectiveness of GnRH-a to resist endometrial cancer.2. The expression of C-myc protein exists in HEC-1B cells;and the PTEN protein is mssing. It clues that C-myc and PTEN plays an important role in endometrial cancer development .3. Triptorelin acetate can change HEC-1B cell morphology significantly.4. Triptorelin acetate can change the expression of C-myc and PTEN,and there is a negative correlation between the two.It clues that triptorelin acetate can reduced the expression of C-myc and increase the expression of PTEN to inhibit the endometrial cancer cell proliferation.