CDC28蛋白激酶调节亚基2对卵巢癌A2780细胞丝足形成的影响及其机制研究  

CKS2 affects filopodia formation of A2780 cells through regulating CDC42 alternative splicing

作　　者：孙倩玲 董梦梦 易祎[1] 胡锦辉[1] 张米妮 朱琳[1] 黄石榴 孙贤青[1] 袁成福[1] 

机构地区：[1]三峡大学医学院生物化学与分子生物学教研室,湖北宜昌443002

出　　处：《解放军医学杂志》2016年第8期618-623,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81550029;30800410);三峡大学人才启动基金(KJ2014B064);湖北省自然科学基金(2015CFB198);肿瘤微环境与免疫治疗湖北省重点实验室(三峡大学)开放基金课题(2015KZL02);宜昌市科学技术局项目(A16-301-31)~~

摘　　要：目的探讨CDC28蛋白激酶调节亚基2(CKS2)对A2780细胞丝足形成的影响及其可能机制。方法采用慢病毒介导的sh RNA敲除A2780细胞的CKS2基因,显微镜下观察细胞丝足的变化;采用细胞划痕实验检测A2780细胞迁移能力的变化;采用real time PCR检测CKS2敲除对CDC42两种剪接变异体(CDC42-V1和CDC42-V2)表达的影响,以及不同卵巢癌标本中CKS2、CDC42-V1和CDC42-V2剪接变异体的表达情况。结果 CKS2表达抑制后,A2780细胞丝足明显减少,细胞迁移能力明显减弱(P<0.05),CDC42-V1 m RNA表达降低,而CDC42-V2 m RNA表达升高(P<0.05)。Real time PCR结果显示,卵巢癌组织标本中CKS2及CDC42-V1的表达高于对应的正常卵巢组织,而CDC42-V2的表达低于对应的正常卵巢组织(P<0.05)。结论 CKS2通过调控CDC42的选择性剪接而影响细胞丝足的形成,进而影响卵巢癌细胞A2780的迁移能力。Objective To study the effect of CKS2 on filopodia formation of A2780 cells through regulating CDC42 alternative splicing. Methods The filopodia were observed after CKS2 of A2780 cells were knocked down by lentivirus-mediated sh RNA; the migrating ability of A2780 cells was also measured by wound healing assay; the expression levels of splicing variants CDC42-V1 and CDC42-V2 m RNA was determined by real time PCR after CKS2 knockdown. The expression levels of CKS2, CDC42-V1 and CDC42-V2 m RNA were further measured in each ovarian cancer and normal ovarian samples in the same way. Results The filopodia on A2780 cells obviously decreased, and the migrating ability of A2780 cells was also remarkably reduced after CKS2 knockdown（P〈0.05）. Meanwhile the expression of CDC42-V1 m RNA decreased and CDC42-V2 m RNA increased after CKS2 knockdown（P〈0.05）. Real time PCR results showed that the m RNA expression levels of CKS2 and CDC42-V1 were higher in ovarian cancer samples than in normal ovarian tissues; however, the expression level of CDC42-V2 m RNA was lower in ovarian cancer samples than in normal ovarian tissues（P〈0.05）. Conclusion CKS2 may affect the filopodia formation of A2780 cells through regulating CDC42 alternative splicing, and further affect the migrating ability of A2780 cells.

关 键 词：CDC28蛋白激酶调节亚基2 CDC42 GTP结合蛋白质 卵巢肿瘤 伪足 选择性剪接 

分 类 号：R737.31[医药卫生—肿瘤]