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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 investigate the effect of CDK28 protein kinase regulatory subunit 2 (CKS2) on the formation of silk foot in A2780 cells and its possible mechanism. Methods The CKS2 gene of A2780 cells was knocked out by lentivirus-mediated sh RNA. The changes of cell line footprints of A2780 cells were observed under microscope. The migration of A2780 cells was detected by cell scratch assay. The real- Splicing variants (CDC42-V1 and CDC42-V2), as well as the expression of CKS2, CDC42-V1 and CDC42-V2 splicing variants in different ovarian cancer samples. Results After CKS2 expression was inhibited, A2780 cells reduced significantly in silky foot and cell migration ability was significantly decreased (P <0.05), CDC42-V1 m RNA expression decreased, while CDC42-V2 m RNA expression increased (P <0.05). Real time PCR results showed that the expression of CKS2 and CDC42-V1 in ovarian cancer specimens was higher than that in normal ovarian tissues, while the expression of CDC42-V2 was lower than that in normal ovarian tissues (P <0.05). Conclusion CKS2 affects the formation of cell line footprints by regulating the alternative splicing of CDC42, which in turn affects the migration ability of ovarian cancer cell line A2780.