Role Of GPER And Its DNA Hydroxymethylation Regulation In Insulin Induced Abnormal Endometrial Proliferation

Objective: To explore the effect of Insulin in the expression level of GPER protein in endometrial cancer cell line Hec- 1 a and Ishikawa, and to determine the influence of interference GPER protein expression in endometrial cancer cell proliferation ability. Methods: The effect of different concentrations of insulin in endometrial cancer cells Hec- 1 a and Ishikawa proliferation was detected by Western blotting and SRB method; rt-PCR was used for screening the corresponding receptor of insulin influence in endometrial cancer cells; Western blotting and confocal immunofluorescence test was used to confirm the influence of Insulin in GPER expression; GPER-si RNA was designed, synthesized and transiently transfected to Hec-1a and Ishikawa cells; Western blotting was also used for detect the efficacy of transfection and the influence of GPER down regulation to downstream pathway; endometrial cancer cell proliferation ability changes was detected by SRB test. Results: Insulin promoted endometrial cancer cells Hec- 1 a and Ishikawa proliferation ability by regulating AKT/PI3 K pathway phosphorylation level; insulin also increase endometrial cancer cells sensitivity to estrogen by raising GPER expression level; GPER was involved in the process of insulin regulating endometrial cancer cell Hec- 1 a and Ishikawa proliferation. Conclusion: Insulin could increase endometrial cancer cells sensitivity to estrogen by increasing the expression of GPER protein, therefore regulating AKT/PI3 K pathway phosphorylation level, promoting endometrial cancer cell proliferation. In conclusion, insulin plays crucial role in the occurrence and development of endometrial carcinoma.Objective: To explore the effect of Insulin in the expression level of TET1 protein in endometrial cancer cell line Hec- 1 a and Ishikawa, and to determine the influence of interference TET1 protein expression in endometrial cancer cell proliferation ability. Methods: Western blotting and confocal immunofluorescence test was used to confirm the influence of Insulin in TET1 expression; TET1-si RNA was designed, synthesized and transiently transfected to Hec-1a and Ishikawa cells; Western blotting was also used for detect the efficacy of transfection and the influence of TET1 down regulation to downstream pathway; endometrial cancer cell proliferation ability changes was detected by SRB test. Results: Insulin could increase the expression of TET1 protein, therefore regulating AKT/PI3 K pathway phosphorylation level, promoting endometrial cancer cell proliferation. Conclusion: TET1 was involved in the progression of insulin regulating endometrial cancer cell Hec- 1 a and Ishikawa proliferation. In conclusion, insulin plays crucial role in the occurrence and development of endometrial carcinoma.Objective: To investigate TET1(Ten-Eleven Translocation 1) expression in different kinds of endometrial hyperplasia and endometrial cancer and the effects of TET1 gene expression on biological behavior of endometrial cancer Hec-1a and Ishikawa cells. Methods: The expression of TET1 was determined by immunohistochemistry; TET1-si RNA was designed, synthesized and transiently transfected to Hec-1a and Ishikawa cells; the efficacy of transfection was tested by Western-blotting; the influence of TET1 knock-down on proliferation, migration, invasion and cell cycle was detected by SRB, wound scratch assay, transwell assay and flow cytometry. Results: The expression of TET1 protein increased significantly along with the development of endometrial hyperplasia and peaked in endometrial cancer tissues. The down-regulation of TET1 inhibited the proliferation, migration and invasion of endometrial cancer cell lines. Cells with TET1 gene knock-down showed cell cycle arrest at G1/S phase. Conclusion: The expression of TET1 is associated with the development of endometrial hyperplasia and knock-down TET1 in endometrial cancer cell lines affects proliferation, migration and invasion.Objective: Explore the influence of insulin in GPER promoter regions hydroxylmethylation level in endometrial cancer cells and the role of TET1 under this regulation. Methods: The expression of GPER and TET1 in different types of endometrial tissue and their correlation was determined by serial section of tissue microarray and immunohistochemical stain; Western blotting, luciferase reporter gene test and Hme Dip was used to detect the regulation of TET1 to GPER promoter regions activity. Results: TET1 and GPER expression in different pathological type of endometrial tissue were positively correlated and TET1 can raise the activity of GPER promoter regions through hydroxymethylation to increase the expression of GPER in endometrial cancer cells. Conclusion: TET1 regulates the expression of GPER through adjusting the hydroxylmethylation level of its promoter zone therefore affects endometrial cancer cell proliferation.