丙戊酸对体外培养上皮性卵巢癌细癝KOV3细胞增殖的影响

目的:研究丙戊酸(valproic acid,VPA)对人卵巢癌细胞株SKOV3细胞增殖以及血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶2、9(matrix metallo-proteinase-2、9,MMP-2、9)和上皮性钙黏附素(E-cadherin)表达的影响。方法:将VPA以不同浓度(0、1、2、3、4和5mmol/L)、不同作用时间(24、48和72h)作用于SKOV3细胞,在相差显微镜下观察细胞形态的变化;用四甲基偶氮唑蓝(MTT)比色法测定细胞增殖;用流式细胞仪测定细胞周期动力学变化;用免疫细胞化学方法检测VEGF、MMP-2、MMP-9和E-cadherin蛋白表达情况;用流式细胞仪测定SKOV3细胞VEGF、MMP-2、MMP-9和E-cadherin蛋白含量。结果:1)3mmol/L作用48h后SKOV3细胞形态出现明显变化。2)不同浓度(0、1、2、3、4和5mmol/L)VPA作用不同时间(24、48和72h),作用72h时,细胞抑制率分别为1.08%、5.41%、18.92%、32.70%和48.65%,呈现明显的剂量依赖关系,差异有统计学意义,P=0.004;不同浓度(0、1、2、3、4和5mmol/L)VPA作用不同时间(48和72h)后,首先引起细胞周期分布的改变,随剂量加大和用药时间延长,凋亡率逐渐升高。3)3mmol/L VPA作用48h后与对照组相比,SKOV3细胞质中VEGF、MMP-2和MMP-9棕色颗粒减少,E-cadherin蛋白棕色颗粒增多。SK-OV3细胞VEGF、MMP-2和MMP-9蛋白含量明显减少,E-cadherin蛋白含量明显增多。结论:VPA抑制SKOV3细胞生长,其作用机制可能与其下调VEGF、MMP-2、MMP-9表达和上调E-cadherin表达有关。

Effects of valproic acid on proliferation in ovarian cancer cell line SKOV3 cells in vitro

OBJECTIVE:To investigate the effects of valproic acid(VPA)on proliferation and expression of vascular endothelial growth factor(VEGF),matrix metalloproteinase-2,9(MMP-2,MMP-9)and E-cadherin in ovarian cancer cell line SKOV3.METHODS:Cultured SKOV3 was treated by VPA with different concentrations and time.The morphological changes of SKOV3 treated by VPA was observed under an inverted microscope.Methyl thiazolyl tetrazolium(MTT)assay was applied to evaluate the cell proliferation.By using immunohistochemistry,the expressions of VEGF,MMP-2,MMP-9 and E-cadherin were assessed in SKOV3.The levels of expression of VEGF,MMP-2,MMP-9 and E-cadherin were determined by using flowcytometry.RESULTS:There was an obvious change in shape and an obvious alter of cell proliferation on SKOV3 during treated with different concentrations and time.The growth of SKOV3 cells was inhibited evidently in vitro.After using VPA for 72h,the cell inhibition rates were 1.08%,5.41%,18.92%,32.70% and 48.65% respectively.The proliferation of SKOV3 cells was inhibited by VPA in a dose and time dependent fashion(P=0.004).The flowcytometric analysis showed that after using VPA(0,1,2,3,4 and 5 mmol/L),the cell cycle distribution changed at first and as the time prolonged the apoptotic peak went higher.The expressions of VEGF,MMP-2 and MMP-9 proteins decreased and the expression of E-cadherin protein increased with the VPA concentration increasement,and the effect appeared when the concentration was 3 mmmol/L.CONCLUSIONS:VPA inhibits the proliferation of SKOV3 cell line.Downregulation of VEGF,MMP-2,MMP-9 and upregulation of E-cadherin appear to be a mechanism in the growth inhibition of VPA-treated SKOV3 cell line.