Blockade Of ONECUT2 Gene Abrogates Cell Proliferation, Metastasis, And Angiogenesis In Ovarian Cancer

Object: Transcript factor ONECUT2 is involved in proliferation, metastasis and angiogenesis as well as EMT of cancer cells recently. ONECUT2 dysregulation is implicated in ovarian cancer. Purpose of this research is to clarify the effect of ONECUT2 silencing on proliferation, migration, invasion and angiogenesis in ovarian cancer cells.Methods: Detected the expression of OC-2 genes in cell lines by q-PCR and ovarian cancer tissues by western blot. Silenced OC-2’ expression to validate its effect on proliferation, metastasis, invasion and angiogenesis of ovarian cancer cells mechanically by CCK-8, transwell/wound healing, and tube formation assay respectively. Constructed ovarian cancer nude mice model to detect OC-2 silencing’s effect on tumor growth and angiogenesis.Results : 1. OC-2 dysregulation was implicated in ovarian carcinoma cell lines. OC-2’s expression of COV-504 and SKOV3 were relatively high both at m RNA transcription and protein translation level than other ovarian cancer cell lines. We detected the expression of OC-2 of ovarian carcinoma samples from 10 patients and 2 ovarian tissues from normal people. Six of ten ovarian cancer samples presented positive of OC-2 while both two normal ovarian tissue samples presented negative. 2. The mean inhibition rate of si RNA on OC-2 protein translation in COV-504 and SKOV3 cell were 85% and 86%; CCK-8 assay indicated that the highest mean inhibition rate of si RNA on metastasis of COV-504 and SKOV3 cell were 11% and 10% at 48h; Transwell assay proved that the mean inhibition rate of si RNA on metastasis of COV-504 and SKOV3 cell were 87% and 88%;Invasion assay demonstrated that the mean inhibition rate of si RNA on invasion of COV-504 and SKOV3 cell were 71% and 77%; Tube formation assay certified that the mean inhibition rate of si RNA on HUVEC tube formation induced by COV-504 and SKOV3 cell were 61% and 60%; The mean inhibition rate of si RNA on p-Akt and p-Erk were 81% and 47% in COV-504 cell and 91% and 95% in SKOV3 cell; Si RNA silenced OC-2 gene inhibited N-cadherin to 63% and increased E-cadherin to 309% in COV-504 cell and inhibited N-cadherin to 14% and increased E-cadherin to 368% in SKOV3 cell. 3. The inhibition rate of si RNA on tumor weights and MVD were 93% and 78% in ovarian cancer xenograft nude mice model, and tumor volume was significantly less than negative control.Conclusion: OC-2’s dysregulation was implicated in different ovarian cancer cell lines and ovarian carcinoma samples of patients.CCK-8 assay proved that downregulation of OC-2inhibited proliferation of ovarian cancer cell; Transwell assay and wound healing assay indicated that si RNA silenced OC-2 gene to inhibit ovarian cancer cell metastasis; Invasion assay and tube formation assay demonstrated that invasion and angiogenesis inhibited after OC-2 were silenced; Downregulation of OC-2 played a tumor suppressive role partially through a functional AKT/ERK signaling pathway verified by western blot assay. Si RNA silencing OC-2 gene resulted in E-cadherin upregulation and N-cadherin downregulation demonstrated that OC-2 was an EMT related transfector. Si RNA silencing OC-2 gene inhibited tumor growth and angiogenesis in ovarian cancer xenograft nude model.In a conclusion, si RNA silenced OC-2 to inhibit cell proliferation, metastasis, and angiogenesis.