摘要
目的:探讨7-脱氢胆固醇还原酶(DHCR7)在上皮性卵巢癌细胞系SKOV3中的表达及其作用机制。方法:聚合酶链反应(PCR)检测DHCR7抑制剂(AY 9944)是否诱导增加SKOV3细胞中DHCR7基因的表达水平。细胞计数试剂盒(CCK-8)、细胞划痕试验检测AY 9944对SKOV3细胞活性及迁移能力的影响。通过蛋白质印迹法(Western blot)探究AY 9944对SKOV3细胞磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/细胞周期蛋白D1(Cyclin D1)通路的影响。Graph Pad Prism 8.0.1统计软件进行统计处理。结果:1.AY 9944能够诱导DHCR7基因表达上调(t=8.485,P<0.05)。2.AY 9944使SKOV3细胞活性明显低于对照组(12 h F=35.22,24 h F=20.34,48 h F=37.56,P<0.05),细胞迁移能力低于对照组(对照组24 h划痕恢复64.50%,48 h为81.14%,加药组24 h划痕恢复30.08%,48 h时为32.79%,P<0.05)。3.SKOV3细胞中DHCR7表达上调后细胞Akt蛋白水平无明显变化(t=0.527,P>0.05),磷酸化Akt(p-Akt)水平低于对照组(t=4.297,P<0.05);PI3K水平低于对照组(t=4.779,P<0.05),CCND1 mRNA表达水平降低(t=9.095,P<0.05),表达低于对照组。结论:AY 9944抑制SKOV3细胞增殖和迁移能力。AY 9944抑制DHCR7酶活性,同时诱导DHCR7基因的表达,其作用机制可能是AY 9944抑制PI3K/Akt/Cyclin D1信号通路,减少CCND1的表达。
Objective To study the expression and mechanism of 7-dehydrocholesterolΔ7-reductase(DHCR7)in epithelial ovarian cancer cell line SKOV3.Methods qPCR was used to detect whether DHCR7 inhibitor(AY 9944)could induce the increase of DHCR7 gene expression in SKOV3 cells.Cell counting kit-8(CCK-8)and cell scratch test were used to detect the effect of AY 9944 on SKOV3 cell activity and migration ability.The effect of AY 9944 on phosphoinositide 3 kinase(PI3K)/protein kinase B(Akt)/Cyclin D1 pathway of SKOV3 cells was investigated by Western blot.The data are expressed as mean±standard error(±s)and statistically processed by the GraphPad Prism 8.0.1 statistical software.Results AY 9944 can induce up regulation of DHCR7 gene expression(t=8.485,P<0.05).AY 9944 make the SKOV3 cell activity significantly lower than that of the control group(12 h F=35.22,24 h F=20.34,48 h F=37.56,P<0.05),and the cell migration ability lower than that of the control group(in the control group,scratch recovery 64.50%for 24 h,and 81.14%for 48 h,and in experience group,scratch recovery is 30.08%for 24 h,and 32.79%for 48 h,P<0.05).After the up regulation of DHCR7 expression in SKOV3 cells,the protein level of Akt did not change significantly(t=0.527,P>0.05),and the level of phosphorylated Akt(p-Akt)is lower than that of the control group(t=4.297,P<0.05);The level of PI3K is lower than that of the control group(t=4.779,P<0.05),the expression of CCND1 mRNA is lower(t=9.095,P<0.05),and the expression of Cyclin D1 is lower than that of the control group.Conclusion AY 9944 inhibits the proliferation and migration of SKOV3 cells.AY 9944 inhibits DHCR7 enzyme activity and simultaneously induces DHCR7 gene expression.Its mechanism may be that AY 9944 inhibits PI3K/Akt/Cyclin D1 signal pathway and reduces the expression of CCND1.
作者
刘利芬
仲璐婷
朱维培
Liu Lifen;Zhong Luting;Zhu Weipei(Department of Gynaecology,the Second Affiliated Hospital of Soochow University,Suzhou 215004,China;Huai’an Maternal and Child Health Hospital,Huai’an 223002,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第12期2421-2424,共4页
Chinese Journal of Experimental Surgery
基金
江苏省妇幼健康科研项目(F202127)。
