白藜芦醇联合SB203580对宫颈癌细胞凋亡迁移的影响及机制

Effect and mechanism of resveratrol combined with SB203580 on the apoptosis and migration of cervical cancer cells

目的探讨白藜芦醇联合SB203580对宫颈癌细胞凋亡迁移的影响及机制。方法培养人宫颈癌HeLa细胞,CCK8实验检测不同浓度白藜芦醇作用于细胞48 h后对细胞增殖影响,挑选最佳作用浓度。将细胞分为四组,即对照组(A组):不做处理;SB203580组(B组),加10μmol/L SB203580;白藜芦醇组(C组),加60μmol/L白藜芦醇;白藜芦醇组+SB203580组(D组),加60μmol/L白藜芦醇和10μmol/L SB203580。流式细胞仪、Transwell小室检测四组细胞的凋亡率和迁移数,Western blot检测p38、p-p38、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved-caspase3)、B型白细胞/2型淋巴细胞样蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达。结果白藜芦醇能显著抑制人宫颈癌HeLa细胞增殖,60μmol/L白藜芦醇抑制了一半的细胞增殖(P<0.001);B组(14.83±1.23)%细胞凋亡率显著低于A组(3.42±0.78)%,细胞迁移数(189.20±7.93)显著高于A组(135.40±7.23);C组(35.57±1.86)%和D组(23.45±1.45)%细胞凋亡率显著高于A组,C组(51.30±5.49)和D组(97.60±6.88)细胞迁移数显著低于A组;D组细胞凋亡率显著高于B组,低于C组,细胞迁移数显著低于B组,高于C组(P<0.001);B组p-p38[(0.087±0.011比0.152±0.014)]、Cleaved-caspase3 [(0.162±0.011比0.093±0.009)]、Bax蛋白表达[(0.191±0.013)比(0.154±0.012)]显著低于A组,Bcl-2蛋白表达[(0.208±0.014)比(0.347±0.015)]显著高于A组;C组和D组p-p38[(0.282±0.016)、(0.201±0.015)]、Cleaved-caspase3[(0.315±0.013)、(0.223±0.012)]、Bax蛋白表达[(0.441±0.015)、(0.294±0.014)]显著高于A组,C组和D组Bcl-2蛋白表达[(0.071±0.010)、(0.156±0.012)]显著低于A组;D组p-p38、Cleaved-caspase3、Bax蛋白表达显著高于B组,低于C组,Bcl-2蛋白表达显著低于B组,高于C组(P<0.001)。p38蛋白表达在各组之间差异无统计学意义(P>0.05)。结论白藜芦醇通过调控p38MAPK信号通路抑制人宫颈癌HeLa细胞增殖和迁移,并通过调节Cleaved-caspase3、Bax、Bcl-2促进细胞凋亡。

Objective To investigate the effect and mechanism of resveratrol combined with SB203580 on the apoptosis and migration of cervical cancer cells.Methods Human cervical carcinoma HeLa cells were cultured,and the effect of different concentrations of resveratrol on cell proliferation was detected by CCK8 assay,and optimal concentration was selected.The cells were assigned into four groups:control group(group A),no treatment;SB203580 group(group B),with 10μmol/L SB203580;resveratrol group(group C),with 60μmol/L resveratrol;resveratrol and SB203580 group(group D),with 60μmol/L resveratrol and 10μmol/L SB203580.Cell apoptosis and migration number were detected in four groups by flow cytometry and Transwell assay separately;expressions of p38,p-p38,Cleaved cysteinyl aspartate specific proteinase 3(Cleaved-caspase 3),B-cell leukemia/lymphoma 2-like proteins(Bcl-2),Bcl-2 associated X protein(Bax)was detected by Western blot.Results Resveratrol could significantly inhibit the proliferation of human cervical carcinoma HeLa cells,60μmol/L resveratrol inhibited half cell proliferation(P<0.001).Apoptosis rate(14.83±1.23)%in group B was significantly lower than(3.42±0.78)%of group A,the number of cell migration(189.20±7.93)was significantly higher than(135.40±7.23)of group A.Apoptosis rates in group C(35.57±1.86)%and group D(23.45±1.45)%were significantly higher than group A.Cell migration numbers in group C(51.30±5.49)and group D(97.60±6.88)were significantly lower than group A.The apoptosis rate in group D was significantly higher than group B,and lower than group C,and its number of cell migration was significantly lower than group B,and higher than group C(P<0.001).The expressions of Cleaved-Caspase 3,Bax protein and p-p38 in group B were significantly lower than group A[(0.162±0.011)vs.(0.093±0.009),(0.191±0.013)vs.(0.154±0.012),(0.087±0.011)vs.(0.152±0.014),respectively],and the expression of Bcl-2 protein was significantly higher than group A[(0.208±0.014)vs.(0.347±0.015)].The expressions of p-p38,Cleaved-caspase 3,Bax protein in group C[(0.282±0.016),(0.315±0.013)and(0.441±0.015),respectively]and group D[(0.201±0.015),(0.223±0.012)and(0.294±0.014),respectively]were significantly higher than group A,and the expressions of Bcl-2 protein in group C(0.071±0.010)and group D(0.156±0.012)were significantly lower than group A.The expressions of p-p38,Cleaved-caspase 3,Bax protein in group D were significantly higher than group B,but lower than group C.The expression of Bcl-2 protein in group D was significantly lower than group B,but higher than group C(P<0.001).There was no significant difference in p38 protein expression among the groups(P >0.05).Conclusions Resveratrol can inhibit the proliferation and migration of human cervical cancer HeLa cells through regulating the p38MAPK signaling pathway,and promote cell apoptosis through regulation of Cleaved-caspase 3,Bax and Bcl-2.