Effect Of AM On Angiogenic Mimicry Of Ovarian Cancer Cells In Vitro

Objective:Among all types of ovarian cancer,epithelial ovarian cancer（EOC）is the most common.The CAOV₃cell line in EOC was originally isolated and cultured from ovarian cancer in a 54-year-old Caucasian female.This cell line has good stability sex,widely used in in vitro studies of ovarian cancer.Adrenomedullin（AM）is essentially a peptide substance with multiple functions.It was first discovered in pheochromocytoma and has been confirmed to be closely related to the occurrence and development of tumors.Adrenomedullin inhibitors（AM22-52）can play a corresponding role by blocking the specific binding of AM to its receptors.In this study,the CAOV₃line was used as the research object,and the effects of AM and AM22-52 on the angiogenesis mimetic of the cancer cells in vitro were explored through in vitro experiments.Methods:The experiment was divided into four groups:normal control group,AM exposure group,AM22-52 exposure group and AM and AM22-52 co-incubation group.The cck-8 assay was used to detect CAOV₃cells exposed to different concentrations of AM and AM22-52 at different time points,and the optical absorbance（OD）value was measured at 450 nm to detect the effects of AM and AM22-52 on the growth rate of CAOV₃cells.Determine the concentration and time of action of AM and AM22-52;use Matrigel gel to conduct three-dimensional culture experiments,observe the in vitro tube-forming effect of CAOV₃cells under the intervention of AM and AM22-52 drugs;The effects of AM and AM22-52 drug intervention on the migration rate of CAOV₃cells were observed.Results:1.CAOV₃cells were incubated with AM and AM22-52 at four concentrations of 1×10^-9,1×10^-8,1×10^-7and 1×10^-6μmol/L for 24h,48h and 72h,respectively.8 kits were tested,the results showed that compared with the normal control group,AM significantly promoted the proliferation of CAOV₃cells,and there was a statistical difference.The effect of AM22-52 on the proliferation of CAOV₃cells was not obvious,and compared with the normal control group,the difference was not statistically significant;compared with the AM group,the AM and AM22-52co-incubation group could significantly inhibit the induction of AM.The proliferation of CAOV₃cells was statistically different,and it increased with the increase of drug concentration and time,also in a concentration-and time-dependent manner;2.CAOV₃cells were incubated with 1×10^-6μmol/L AM and AM22-52.Afterwards,the results of Matrigel in vitro three-dimensional culture experiments showed that compared with the normal control group,AM could significantly stimulate CAOV₃cells to form tubules.The difference was statistically significant（P<0.05）;AM22-52did not significantly stimulate the ability of CAOV₃cells to form tubes after incubation,and the results were similar to those in the normal control group,with no statistical difference;the co-incubation group could significantly inhibit the formation of tubules and tubules by CAOV₃cells.Compared with the AM exposure group,the number of nodes formed,the number of inter-tubular junctions and the total length of the formed tubules were significantly different（P<0.05）,indicating that AM has a significant effect on the in vitro angiogenesis mimic of ovarian cancer CAOV₃cells.AM22-52can significantly inhibit the promoting effect of AM on tubulogenesis;3.after1×10^-6μmol/L AM and AM22-52 incubated CAOV₃cells for 48h,the cell scratch test results showed that compared with the normal control group,the After AM incubation,the migration ability of CAOV₃cells was significantly improved,while the migration activity of AM22-52 group had little difference compared with the normal control group;Enhance the invasion of CAOV₃cells,AM22-52 can significantly inhibit the invasion of CAOV₃cells.Conclusion:1.AM can significantly promote the proliferation of CAOV₃cells,and AM22-52 can inhibit the proliferation of CAOV₃cells induced by AM;2.AM can significantly promote the angiogenesis mimetic of CAOV₃cells in vitro,and AM22-52can significantly inhibit the proliferation of AM.In vitro angiogenesis mimetic effect;3.AM can significantly improve the cell migration ability of CAOV₃cells,AM22-52can significantly promote the cell migration ability of AM.The above results reveal that AM can significantly stimulate the angiogenesis mimetic effect of CAOV₃cells in vitro,and can also affect their growth and migration ability.Inhibiting AM may be a new idea and method for the potential treatment of epithelial ovarian cancer.