TIPIN分子与肝细胞癌预后的关系及其生物学作用机制

Relationship between TIPIN and prognosis of hepatocellular carcinoma cells and its biological mechanism

目的分析节律分子Timeless(TIM)相互作用蛋白(TIM interacting protein,TIPIN)在肝细胞癌(hepatocellular carcinoma,HCC)组织中的表达及其与患者预后的关系,并探究TIPIN对肝癌细胞增殖的影响及可能的生物学作用机制。方法通过UALCAN数据库分析TIPIN在HCC组织中的表达情况及其与HCC患者预后的关系。将siTIPIN及其阴性对照序列分别转染至肝癌MHCC-97H细胞,采用qRT-PCR和Western blot检测TIPIN的表达情况,采用MTS实验和平板细胞克隆形成实验检测TIPIN表达对细胞增殖的影响。利用UALCAN数据库对TIPIN进行GO和KEGG富集分析,以及TIPIN与TIM、TP53的关联分析。结果UALCAN数据库分析结果显示,TIPIN在HCC组织中的表达明显高于正常肝组织(P<0.01);TIPIN高表达患者的总生存期和无病生存期均明显短于TIPIN低表达患者(P=0.037,0.021)。与正常肝细胞HL7702相比,肝癌细胞MHCC-97H、MHCC-97L、HepG2、BEL-7402中TIPIN的mRNA和蛋白表达水平均明显升高(均P<0.01)。MTS和克隆形成实验结果显示,沉默TIPIN表达后肝癌MHCC-97H细胞的增殖和平板细胞克隆形成能力明显受到抑制(均P<0.05)。GO和KEGG富集分析结果显示,TIPIN可能通过调控细胞分裂、DNA复制、细胞周期的相互作用通路促进肝癌的恶性进程。Pearson相关性分析结果显示,TIPIN分子与TIM表达呈明显正相关(r=0.67,P<0.01)。TIPIN在TP53突变HCC患者中的表达显著高于TP53野生型HCC患者(P<0.01)。结论TIPIN在肝癌中表达上调,且与患者不良预后密切相关,沉默TIPIN可抑制肝癌细胞增殖。TIPIN在肝癌中可能通过与TIM形成复合体或调控TP53信号通路发挥其生物学作用。

ObjectiveTo analyze the expression of rhythmic molecules timeless interacting protein(TIPIN)in hepatocellular carcinoma(HCC)tissues and its relationship with the prognosis of patients,and to explore the effect of TIPIN on the proliferation of HCC cells and its possible biological mechanism.MethodsThe expression of TIPIN in HCC and its relationship with the prognosis of HCC patients were explored through the UALCAN database.The siTIPIN and its negative control sequence were transfected into liver cancer MHCC-97H cells,respectively,and the expression of TIPIN was detected by qRT-PCR and Western blot.The effect of TIPIN expression on cells proliferation was detected by the MTS assay and the plate cell clone formation assay.The GO and KEGG enrichment analysis of TIPIN and its association with Timeless(TIM)and TP53 were performed using UALCAN database.ResultsUALCAN database analysis showed that the expression of TIPIN in HCC tissues was significantly higher than that in normal liver tissues(P<0.01).The overall survival and disease-free survival in patients with high TIPIN expression were significantly lower than those of patients with low TIPIN expression(P=0.037,0.021).Compared with normal liver cells HL7702,the mRNA and protein expression of TIPIN in MHCC-97H,MHCC-97L,HepG2 and BEL-7402 were significantly increased(all P<0.01).MTS and plate cell clonal formation experiments showed that the proliferation and clonogenesis of MHCC-97H cells were significantly inhibited after silencing the expression of TIPIN(all P<0.05).GO and KEGG analysis showed that TIPIN might promote the malignant process of HCC by regulating the interaction pathways of cell division,DNA replication and cell cycle.Pearson correlation analysis showed that TIPIN was positively correlated with TIM expression(r=0.67,P<0.01).The expression of TIPIN in TP53 mutant HCC patients was significantly higher than that in TP53 wild type HCC patients(P<0.01).Conclusions The expression of TIPIN is up-regulated in liver cancer and is closely related to poor prognosis.Silencing TIPIN can inhibit the proliferation of liver cancer cells.TIPIN may play a biological role in liver cancer cells by forming complex with TIM or regulating TP53 signal pathway.