桔皮素对卵巢癌SKOV3细胞的体外作用及作用机制

Effect of Tangeretin on Ovarian Cancer SKOV3 Cells in Vitro and Its Mechanism

目的探讨桔皮素对卵巢癌SKOV3细胞的体外作用,以及对Ras/丝裂原活化的细胞外信号调节激酶(MEK)/细胞外信号调节激酶(ERK)信号通路的影响。方法将卵巢癌SKOV3细胞分为空白对照组(A组,0μmol/L),阳性对照组(B组,75 nmol/L阿霉素),桔皮素低、中、高剂量组(分别为C_(1)组、C_(2)组、C_(3)组,桔皮素2,5,10μmol/L)。取对数生长期的SKOV3细胞,采用CCK-8法检测细胞存活率,采用改良Matrigel Boyden室测定法测定细胞的侵袭性,采用BD FACSCantoⅡ型流式细胞仪检测细胞凋亡率,采用蛋白免疫印迹法检测细胞Ras,MEK,ERK的蛋白表达水平。结果与A组比较,B组、C_(1)组、C_(2)组、C_(3)组卵巢癌SKOV3细胞存活率及Ras,MEK,ERK蛋白表达水平均显著降低,细胞凋亡率均显著升高,侵袭细胞数均显著减少,且C_(1)组、C_(2)组、C_(3)组效应均呈剂量依赖性(P<0.05);但C_(3)组和B组卵巢癌SKOV3细胞上述指标均无显著差异(P>0.05)。结论桔皮素可有效抑制卵巢癌SKOV3细胞的增殖与侵袭,并促进其凋亡,作用机制可能与抑制Ras/MEK/ERK信号通路的蛋白表达有关。

Objective To investigate the effect of tangeretin on ovarian cancer SKOV3 cells in vitro and the Ras/mitogen-activated protein kinase kinase(MEK)/extracellular-signal regulated kinase(ERK)pathway.Methods Ovarian cancer SKOV3 cells were divided into the blank control group(group A,0μmol/L),positive control group(group B,75 nmol/L of doxorubicin),hesperidin low-,medium-and high-dose groups(groups C_(1),C_(2)and C_(3),2,5 and 10μmol/L of tangeretin).CCK-8 assay was used to detect cell survival rate,Matrigel Boyden chamber assay was used to detect cell invasion,and BD FacscantoⅡflow cytometry was used to detect cell apoptosis rate,and Western blot was used to detect the expression levels of Ras,MEK and ERK protein.Results Compared with those in group A,the survival rate of ovarian cancer SKOV3 cells and the expression levels of Ras,MEK and ERK protein of ovarian cancer SKOV3 cells decreased significantly,the apoptosis rate of ovarian cancer SKOV3 cells increased significantly,while the number of invasion cells decreased significantly in groups B,C_(1),C_(2)and C_(3),and the effects in groups C_(1),C_(2)and C_(3)were dose-dependent(P<0.05).However,there was no significant difference in the above indexes of ovarian cancer SKOV3 cells between group C_(3)and group B(P>0.05).Conclusion Tangeretin can effectively inhibit the proliferation and invasion of ovarian cancer SKOV3 cells and promote cell apoptosis.The mechanism may be related to the inhibition of Ras/MEK/ERK signal pathway-related protein expression by tangeretin.