白杨素联合氯喹抑制卵巢癌细胞增殖促进凋亡的研究  

Chrysin inhibits proliferation and promotes apoptosisof ovarian cancer cells by inducing autophagy

作　　者：史于传 何宇 杨阳 范懿隽[1] 杨芳[1] 詹磊[1] 卫兵[1] Shi Yuchuan;He Yu;Yang Yang;Fan Yijun;Yang Fang;Zhan Lei;Wei Bing(Dept of Obstetrics and Gynecology,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601;Dept of Pharmacy,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601)

机构地区：[1]安徽医科大学第二附属医院妇产科,合肥230601 [2]安徽医科大学第二附属医院药剂科,合肥230601

出　　处：《安徽医科大学学报》2022年第4期510-514,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81802586)。

摘　　要：目的探究白杨素联合氯喹对卵巢癌细胞增殖和凋亡的作用及机制。方法选取手术切除后卵巢癌标本和正常卵巢标本,使用免疫组化法检测卵巢癌和正常卵巢组织自噬相关蛋白LC3的表达水平。使用CCK-8法检测卵巢癌细胞SKOV3的增殖。使用Western blot和免疫荧光法检测SKOV3细胞中LC3的表达。使用电镜检测SKOV3细胞中自噬小体的数目。使用流式细胞术检测SKOV3细胞凋亡情况。结果LC3在卵巢癌中的表达量高于其在正常卵巢组织中的表达。分别用0、20、40、60、80、100μmol/L浓度的白杨素处理SKOV3细胞24 h,随着浓度增加,细胞增殖能力降低。40μmol/L白杨素处理SKOV3细胞24 h后,与对照组相比,白杨素上调了LC3的表达(P<0.05)。与对照组比较,白杨素处理24 h的SKOV3细胞后,透射电镜下可以观察到其中间出现较多的自噬小体(P<0.05),在对照组则未观察到自噬小体的存在。当白杨素与CQ联合使用24 h后,联合处理组的LC3蛋白表达量高于白杨素和氯喹单独处理组(P<0.05)。白杨素可以诱导SKOV3细胞的凋亡,且白杨素+氯喹组凋亡比例最高(P<0.05)。结论白杨素能够诱导自噬进而抑制卵巢癌细胞增殖和促进凋亡。Objective To explore the effect and mechanism of chrysin and chloroquine(CQ)on the proliferation and apoptosis of ovarian cancer cells.Methods Ovarian cancer specimens and normal ovarian specimens were selected after surgical resection and immunohistochemical experimental methods was used to detect the expression level of autophagy-related protein LC3 in ovarian cancer and normal ovarian tissues.The CCK-8 method was used to detect the proliferation of ovarian cancer cell SKOV3.Western blot and immunofluorescence were used to detect the expression of LC3 in SKOV3 cells.An electron microscope was used to detect the number of autophagosomes in SKOV3 cells.Flow cytometry was used to detect SKOV3 cell apoptosis.Results The expression level of LC3 in ovarian cancer was higher than that in normal ovarian tissues.SKOV3 cells were treated with chrysin at concentrations of 0,20,40,60,80,100μmol/L for 24 hours.As the concentration increased,cell proliferation decreased significantly.Compared with the control group,40μmol/L,chrysin significantly up-regulated the expression of LC3 after 24 hours(P<0.05).Compared with the control group,after 24 hours of SKOV3 cells treated with chrysanthemum,more autophagosomes were observed under transmission electron microscope(P<0.05),but no autophagosome was observed in the control group.When chrysin and chloroquine were used in combination for 24 hours,the expression of LC3 protein in the combined treatment group was higher than that in the chrysin and CQ treatment group(P<0.05).Chrysin could obviously induce apoptosis of SKOV3 cells,and the group of Chrysin+CQ showed the highest proportion of apoptosis(P<0.05).Conclusion Chrysin can induce autophagy to inhibit the proliferation of ovarian cancer cells and promote apoptosis.

关 键 词：白杨素 SKOV3细胞 自噬 增殖 凋亡 

分 类 号：R711.75[医药卫生—妇产科学]