PLK2协同p21影响骨肉瘤细胞增殖的作用机制

The mechanism of PLK2 synergistic p21 affecting the proliferation of osteosarcoma cells

目的通过实验研究PLK2协同p21影响骨肉瘤疾病发生发展的潜在作用机制。方法从GEO数据库获取骨肉瘤组织与正常组织的lncRNA差异化表达;准备HEK293T细胞和U2OS、Sao-2人骨肉瘤细胞系进行培养,利用siRNA技术在U2OS细胞系中抑制PLK2的表达并实施敲低试验,从U2OS细胞中分离细胞总RNA进行实时定量聚合酶链反应,对U2OS细胞系实施免疫印迹(IB)和免疫沉淀(IP)分析,再对其进行EdU实验和细胞活力测定,最后使用SPSS 17.0版本进行统计学分析,将癌组织与癌旁正常组织进行比较。结果与正常组织相比,PLK2在骨肉瘤组织中表达更低;在构建的两种敲低PLK2的小干扰RNA和携带PLK2的重组质粒转染到U2OS细胞实验中,PLK2敲除后发现U2OS细胞的增殖能力增强,而PLK2过表达则相反。在U2OS细胞中检测PLK2和p21的体内相互作用,发现p21的蛋白水平与PLK2的蛋白水平变化一致且差异具有统计学意义。结论PLK2参与了抑制骨肉瘤细胞增殖的过程,可能是通过与p21相互作用而发生的。

Objective To study the potential mechanism of PLK2 and p21 affecting the occurrence and development of osteosarcoma.Methods The differential expression of lncRNA in osteosarcoma tissues and normal tissues was obtained from the GEO database.HEK293 T cells and U2OS and Sao-2 human osteosarcoma cell lines were prepared for culture.The expression of PLK2 in U2OS cell lines was inhibited by siRNA technology and knockdown assay was performed.Total RNA was isolated from U2OS cells for real-time quantitative polymerase chain reaction.Immunoblotting(IB)and immunoprecipitation(IP)analysis were performed on U2OS cell lines,followed by EdU assay and cell viability assay.Finally,SPSS 17.0 version was used for statistical analysis.Cancer tissues were compared with adjacent normal tissues.Results Compared with normal tissues,PLK2 was lower in osteosarcoma tissues.In the experiment of transfecting two kinds of small interfering RNA knocking down PLK2 and recombinant plasmid carrying PLK2 into U2OS cells,it was found that the proliferation ability of U2OS cells was enhanced after PLK2 knockout,while PLK2 overexpression had the opposite effect.The in vivo interaction between PLK2 and p21 was detected in U2OS cells,and it was found that the protein level of p21 was consistent with the protein level of PLK2 and was statistically significant.Conclusion PLK2 is involved in the process of inhibiting the proliferation of osteosarcoma cell,which may occur by interacting with p21.