基于数据挖掘分析软组织肉瘤中PRC1的表达与意义及其对肉瘤细胞生长的影响

Expression and clinical significance of PRC1 in soft tissue sarcoma based on data mining and effect of PRC1 on the growth of liposarcoma

目的通过数据库挖掘,深入探讨细胞分裂蛋白调节因子1(PRC1)在软组织肉瘤(STS)中的表达及意义,以及沉默PRC1对脂肪肉瘤(LPS)细胞增殖和周期的影响。方法Oncomine数据库提取PRC1在STS组织和癌旁正常组织中的表达数据,随后以TCGA数据库加以验证;OncoLnc数据库提取PRC1的预后信息,分析PRC1表达水平与STS患者预后的关系;CCLE数据库提取PRC1在STS细胞中的表达数据;String网站获取PRC1的共表达分子并绘制PRC1的共表达网络。Western blotting和Real-time PCR检测LPS细胞株SW872及人皮下前脂肪细胞株HPA-s中PRC1的表达;应用siRNA构建沉默PRC1的SW872细胞(SW872-siPRC1),MTT和流式细胞术分别检测沉默PRC1后对SW872增殖及细胞周期的影响。结果Oncomine数据库中共11项研究涉及STS组织和癌旁正常组织中PRC1的表达,PRC1在STS中高表达(P<0.05)。TCGA数据库的验证结果与Oncomine数据库挖掘结果一致。OncoLnc数据库的预后分析结果显示,高表达PRC1的STS患者总体生存率较差(P<0.05)。CCLE数据库的结果提示,PRC1在STS细胞中呈普遍高表达(P<0.05)。String网站绘制PRC1的共表达网络,包括11个节点和55个连接。PRC1在LPS细胞SW872中呈高表达,细胞增殖曲线显示,沉默PRC1的SW872-siPRC1细胞培养48、72 h,较癌旁对照组SW872-NC细胞增殖明显降低,差异具有统计学意义(P<0.05)。与SW872-NC细胞比较,SW872-siPRC1组细胞G1期的细胞比例明显下降[(40.27±7.42)%vs.(62.01±4.89)%];G2/M期的细胞比例明显上升[(25.65±1.54)%vs.(8.17±0.96)%],差异均有统计学意义(均P<0.05)。结论PRC1在STS组织和细胞中呈现高表达,与STS预后相关;沉默PRC1基因抑制LPS SW872细胞增殖,阻滞细胞于G2/M期,可能为STS特别是LPS的临床治疗和预后研究提供潜在靶点。

Objective To investigate the expression and clinical significance of protein regulator of cytokinesis 1(PRC1)in soft tissue sarcoma(STS)and the effects of down-regulating the expression of PRC1 on the proliferation and cell cycle of human liposarcoma cell by data mining.Methods Oncomine database was used to analyze the expression of PRC1 in STS tissue and normal tissue,which was then verified by TCGA database.The prognosis data downloaded from OncoLnc database were used to analyze the correlation between PRC1 expression and prognosis of STS patients.Meanwhile,to collect PRC1 expression in STS cells,we used publicly available data from the Cancer Cell Line Encyclopedia(CCLE)projects.String database was used to determine the co-expression molecules with PRC1 and map the gene co-expression network.The expressions of PRC1 in liposarcoma cell line SW872 and human subcutaneous preadipocytes(HPA-s)were detected by Western blotting and Real-time PCR.PRC1 was silenced in SW872 cell(SW872-siPRC1)by PRC1 target siRNA with SW872-NC cell as its control.MTT assay was used to detect the proliferation of SW872-siPRC1 and SW872-NC cells;flow cytometry was used to detect the cell cycle.Results In the Oncomine database,11 studies involved PRC1 expression in STS tissues and normal tissues.Compared with that of the control,the expression of PRC1 in STS tissues was significantly higher(P<0.05).The results were consistent with those in the TCGA database.The analysis using Oncomine database showed that the high expression level of PRC1 was associated with shorter overall survival(P<0.05).The analysis using CCLE database showed high expression of PRC1 in STS cells(P<0.05).The co-expression network of PRC1 was established by String database,including 11 nodes and 55 connections.PRC1 was over-expressed in liposarcoma cell line SW872.Cell proliferation curve showed that compared with that of SW872-NC cells in the control group,the proliferation of SW872-siPRC1 cells decreased significantly after 48 h and 72 h culture(P<0.05).Compared with SW872-NC cell in the control group,the G1 cell proportion of SW872-siPRC1 cells was(40.27±7.42)%,significantly lower than that of SW872-NC cells(62.01±4.89)%.The G2/M cell proportion of SW872-siPRC1 cells was(25.65±1.54)%,which was significantly higher than that of SW872-NC cells(8.17±0.96)%(both P<0.05).Conclusion Tumor gene database mining shows that PRC1 is highly expressed in STS tissues and STS cells,which is associated with the patient's prognosis.Silencing PRC1 gene can inhibit the proliferation of liposarcoma SW872 cells and keep the cells staying in G2/M phase.PRC1 plays a role in promoting liposarcoma,which may provide a potential target for the clinical treatment and prognosis of soft tissue sarcoma,especially liposarcoma.