Experimental Research On Divalent Cation Modulate SKOV3 Ovarian Cancer Cell Adhesion, Invasion And Metastasis

Objective:To investigate the effects of divalent cation modulate SKOV3 ovarian cancer cell adhesion, invasion and metastasis.Methods:Divide different concentrations of divalent cations (Ca2+, Zn2+, Mn2+) into:(1) Control group, (2) Mn2+(0.25mM) group, (3) Ca2+(2.5,5.0, 10.0mM) group, (4) Mn2+(0.25mM)+Ca2+(2.5,5.0, 10.0mM) group, (5) Zn2+(1.0, 10.0mM) group, (6) Mn2+(0.25mM)+Zn2+(1.0, 10.0mM) group, and roled in the SKOV3 ovarian cancer cells, respectively carryed on the adhesion, invasion and metastasis experiments.1. Adhesion experiment:96 well plate joined in Matrigel, after inoculated cells, being roled in 30min by divalent cations, cell morphological changes were observed with optical microscope, cell counting, and calculated cell adhesion rate.2. Invasion experiment:Transwell joined in Matrigel, after inoculated cells, being roled in 24h by divalent cations, ELIASA detected the OD value of invasion cells in 570 nm, and calculated cell invasion rate.3. Mmetastasis experiment:Cells inoculateed in Transwell, after being roled in 24h by divalent cations, ELIASA detected the OD value of metastasis cells in 570 nm, and calculated cell metastasis rate.Results:1. Adhesion experiment:SKOV3 ovarian cancer cells cultured 30 minutes under the action of divalent cations, evenly distributed in 96 well plate, the same size, form into a round or short spindle, stained nuclear deeply, poor light transmission. At Ca2+ (2.5-10mM) group, adherent cells decreased with Ca2+ concentration increasing, adherence inhibition rate increased from 10.2±7.3% to 62.2±4.7% range (n=3, P<0.05). At Mn2+ (0.25mM)+Ca2+ (2.5-10mM) group, compared with the Mn2+ (0.25mM) control group, adherent cells were significantly reduced, the maximal inhibition concentration was lOmM (65.3±3.8%, n=3, P<0.05). Zn2+ concentration at 1mM, adherent cells increased slightly; at lOmM, the number of adherent cells significantly decreased, adherence inhibition was 76.1±2.3%. At Mn2+ (0.25mM)+Zn2+(1-10mM) group, compared with the Mn2+(0.25mM) control group, Zn2+ concentration at lOmM, adherent cells was significantly reduced, the maximal inhibition concentration was 10mM (71.6±1.9%, n=3, P<0.05).2. Invasion experiment:SKOV3 ovarian cancer cells cultured 24 hours under the action of divalent cations, adhered to the lower chamber surface of transwell after through the matrigel, evenly distribution, the same size, form into a round or irregularity, stained nuclear deeply, poor light transmission. At Ca2+(2.5-10mM) group, invasive cells decreased with Ca2+ concentration increasing, ELIASA detected the absorbance decreased gradually, invasive inhibition rate increased from 24.8±3.8% to 63.2±1.2% range (n=3, P<0.05). At Mn2+(0.25mM)+Ca2+(2.5-10mM) group, compared with the Mn2+(0.25mM) control group, invasive cells were significantly reduced, the maximal inhibition concentration is 10mM (72.8±1.1%, n=3, P<0.05). At Zn2+(1-lOmM) group, with Zn2+ concentration increasing, invasive cells decreased gradually, ELIASA detected the absorbance decreased gradually, invasive inhibition rate increased from 66.0±2.2% to 87.9±1.2% range (n=3, P<0.05). At Mn2+(0.25mM)+Zn2+(1-10mM) group, compared with the Mn2+(0.25mM) control group, invasive cells were significantly reduced, the maximal inhibition concentration was 10mM (90.8±0.9%, n=3, P<0.05).3. Mmetastasis experiment:SKOV3 ovarian cancer cells cultured 24 hours under the action of divalent cations, adhered to the lower chamber surface of transwell, evenly distribution, the same size, form into a round or irregularity, stained nuclear deeply, poor light transmission. At Ca2+(2.5-10mM) group, metastatic cells decreased with Ca2+ concentration increasing, ELIASA detected the absorbance decreased gradually, metastatic inhibition rate increased from 22.4±3.3% to 55.5±5.5% range (n=3, P<0.05). At Mn2+(0.25mM)+Ca2+(2.5-10mM) group, compared with the Mn2+(0.25mM) control group, metastatic cells were significantly reduced, the maximal inhibition concentration is lOmM (75.6±3.3%, n=3, P<0.05). At Zn2+ (1-10mM) group, with Zn2+ concentration increasing, metastatic cells decreased gradually, ELIASA detected the absorbance decreased gradually, metastasis inhibition rate increased from 73.2±1.3% to 91.7±0.2% range (n=3, P<0.05). At Mn2+ (0.25mM)+Zn2+(1-10mM) group, compared with the Mn2+(0.25mM) control group, metastatic cells were significantly reduced, the maximal inhibition concentration is 10mM (93.4±0.9%, n=3, P<0.05).Conclusion:SKOV3 ovarian cancer cells were affected by divalent cation concentration in vitro, Ca2+ and Zn2+ could inhibit cell adhesion, invasion and metastasis functions, meanwhile could significantly reduce the Mn2+ promoting functions of cell adhesion, invasion and metastasis. These results suggest that wash the surgical site with proper divalent cation solutions might diminish perioperative ovarian cancer cell implantation.