摘要
目的:研究caveolin-1脚手架结构域(caveolin-1 scaffolding domain,CSD)多肽对卵巢癌SKOV3细胞迁移和侵袭能力的影响,并探讨其分子机制。方法:用细胞免疫荧光法检测CSD多肽在SKOV3细胞中的渗透情况,划痕实验检测细胞的迁移能力,Transwell细胞侵袭实验检测细胞的侵袭能力,用细胞免疫荧光和蛋白免疫印迹法检测E-cadherin和Vimentin蛋白的表达,蛋白免疫印迹法检测SKOV3细胞中EGFR、PI3K、Akt蛋白的相对磷酸化水平。结果:细胞免疫荧光显示CSD多肽能够顺利进入SKOV3细胞内,与空白对照组相比,CSD多肽能明显抑制SKOV3细胞的迁移能力(P<0.01)和侵袭能力(P<0.01),并且CSD多肽能增加细胞中E-cadherin蛋白水平(P<0.01)和下调Vimentin蛋白的表达(P<0.01);SKOV3细胞用CSD多肽处理后,EGFR、PI3K、Akt蛋白的相对磷酸化水平明显低于空白对照组(P<0.01,P<0.05,P<0.05)。结论:CSD多肽在体外能抑制SKOV3细胞的侵袭和迁移能力,逆转上皮-间质转化表型,EGFR/PI3K/Akt信号通路活性水平的下降可能是其重要的分子机制。
Objective:To study the effect of caveolin-1 scaffolding domain(CSD)polypeptide on the ability of migration and invasion in SKOV3 cell,and to explore its molecular mechanism.Methods:Immunocytofluorescence was applied to observe the infiltrability of CSD polypeptide in SKOV3 cell.The scratch test was applied to test the migratory ability,and Transwell in vitro invasion assay was used for the invasive ability of SKOV3 cell.E-cadherin and Vimentin protein expression were evaluated by Western blot and Immunocytofluorescence.EGFR,PI3K and Akt protein expresion and their relative phosphorylation levels were detected by Western blot.Results:Immunocytofluorescence results showed CSD polypeptide can be detected in cytoplasm of SKOV3 cell.Treated with CSD polypeptide,the ability of invasion and migration were significantly inhibited compare with blank control group(P<0.01).Compared with blank control group,CSD polypeptide can upregulate E-cadherin protein level(P<0.01)and inhibite Vimentin protein expression(P<0.01)in SKOV3 cell.According to Western blot results,the relative phosphorylation levels of EGFR,PI3K and Akt in CSD polypeptide group were significantly lower than that in blank control group(P<0.01,P<0.05,P<0.05).Conclusion:caveolin-1 scaffolding domain polypeptide is able to inhibit the migratory and invasive ability of SKOV3 cell and reverse the epithelial-mesenchymal transition,EGFR/PI3K/Akt signaling pathway may be involved in its molecular mechanisms.
作者
郑庆玲
张婷
张明梅
顾栋桦
ZHENG Qingling;ZHANG Ting;ZHANG Mingmei;GU Donghua(Medicine School of Huzhou University,Zhejiang Huzhou 313000,China;Department of Pathology,Affiliated Suzhou Science & Technology Town Hospital of Nanjing Medical University,Jiangsu Suzhou 215153,China)
出处
《现代肿瘤医学》
CAS
北大核心
2022年第2期201-206,共6页
Journal of Modern Oncology
基金
浙江省湖州市自然科学资金(编号:2020YZ03)
湖州师范学院校级科研项目(编号:2020XJKJ10)。
