女贞子多糖通过EGFR/MAPK信号通路对卵巢癌细胞增殖、凋亡的影响

Influences of Ligustrum lucidum polysaccharide on proliferation and apoptosis of ovarian cancer cellsthrough EGFR/MAPK signaling pathway

目的探究女贞子多糖通过表皮生长因子受体(epidermal growth factor receptor,EGFR)/有丝分裂原活性蛋白激酶(mitogen-activated protein kinase,MAPK)通路对卵巢癌细胞增殖、凋亡的影响。方法取对数生长期的卵巢癌HO8910细胞,将其分为女贞子多糖低剂量(1μg/mL)、中剂量(5μg/mL)、高剂量(10μg/mL)组、pcDNA-EGFR组(转染pcDNA-EGFR)、女贞子多糖+pcDNA组(转染pcDNA和10μg/mL女贞子多糖共同处理)、女贞子多糖+pcDNA-EGFR组(转染pcDNA-EGFR和10μg/mL女贞子多糖共同处理)。甲基噻唑蓝(methyl thiazolyl tetrazolium,MTT)法检测细胞增殖;流式细胞术检测细胞凋亡;Western blot法检测EGFR/MAPK通路蛋白磷酸化水平、B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)表达。结果与对照组相比,女贞子多糖低、中、高剂量组光密度(optical density,OD)值、Bcl-2蛋白、EGFR/MAPK通路蛋白磷酸化水平降低,细胞凋亡率、Bax蛋白表达升高,且呈剂量依赖性(P<0.05);与对照组相比,pcDNA-EGFR组OD值、Bcl-2蛋白、EGFR/MAPK通路蛋白磷酸化水平升高,细胞凋亡率、Bax蛋白表达量显著降低(P<0.05);与女贞子多糖+pcDNA组相比,女贞子多糖+pcDNA-EGFR组OD值、Bcl-2蛋白、EGFR/MAPK通路蛋白磷酸化水平升高,细胞凋亡率、Bax蛋白表达量显著降低(P<0.05)。结论女贞子多糖可能通过抑制EGFR/MAPK信号通路,进而抑制HO8910细胞增殖,促进细胞凋亡。

Objective To explore the effect of Ligustrum lucidum polysaccharide on the proliferation and apoptosis of ovarian cancer cells through the epidermal growth factor receptor(EGFR)/mitogen-active protein kinase(MAPK)pathway.Methods Ovarian cancer HO8910 cells in logarithmic growth phase were taken and treated with different doses of Ligustrum lucidum polysaccharide(1μg/mL,5μg/mL,10μg/mL)for 48 h,and cell proliferation was detected by MTT method;cell apoptosis was detected by flow cytometry;the phosphorylation level of EGFR/MAPK pathway protein,B-cell lymphoma-2(Bcl-2)and Bcl-2-related X protein(Bax)expression were detected by Western blot.The pcDNA-EGFR was transfected into HO8910 cells,and the effect of overexpression of EGFR on the proliferation and apoptosis of HO8910 cells was observed;HO8910 cells were treated with 10μg/mL Ligustrum lucidum polysaccharide and pcDNA-EGFR was added to observe the changes in the proliferation and apoptosis of HO8910 cells.Results Compared with the control group,the OD value,Bcl-2 protein,EGFR/MAPK pathway protein phosphorylation levels,cell apoptosis rate were decreased in the Ligustrum lucidum polysaccharide group(1μg/mL,5μg/mL,10μg/mL),the Bax protein expression was increased,and it was dose-dependent(P<0.05);compared with the control group,the OD value,Bcl-2 protein and phosphorylation level of EGFR/MAPK pathway protein were increased,and the apoptosis rate and Bax protein expression were significantly decreased in the pcDNA-EGFR group(P<0.05);compared with the Ligustrum lucidum polysaccharide+pcDNA group,the OD value,Bcl-2 protein and phosphorylation level of EGFR/MAPK pathway protein were increased,and the apoptosis rate and Bax protein expression were significantly decreased in the Ligustrum lucidum polysaccharide+pcDNA-EGFR group(P<0.05).Conclusion Ligustrum lucidum polysaccharide may inhibit the proliferation and promote cell apoptosis of HO8910 cells by inhibiting the EGFR/MAPK signaling pathway.