磷酸化蛋白p27在肝细胞肝癌中表达及其意义

Expression and significance of phosphorylated protein p27 in hepatocellular carcinoma

目的 探讨人肝细胞肝癌组织中磷酸化蛋白p27的表达及其与临床病理特征的关系.方法 利用Ti4＋-IMAC富集和LC-MS2-MS3检测磷酸化蛋白的方法鉴定人肝细胞肝癌组织磷酸化蛋白,进一步利用Western blot检测40例肝细胞肝癌、相应癌旁组织和12例正常肝组织中T187、S10磷酸化p27蛋白（P-p27T187、P-p27S10）的表达.结果在肝癌组织磷酸化蛋白的筛选中鉴定到磷酸化蛋白p27,并确定IPI、磷酸化位点、蛋白激酶等信息,Western blot检测提示P-p27T187与P-p27S10在肝细胞肝癌组织表达明显高于癌旁、正常肝组织,其相对表达量分别为0.635±0.108、0.306±0.078和0.149±0.031,差异有统计学意义（P〈0.05）.且P-p27T187和P-p27S10与肝癌病理分级、临床分期及转移相关,而与发病年龄、性别、肿瘤大小、肿瘤数目等无明显相关（P〉0.05）.结论 磷酸化蛋白p27在肝细胞肝癌中表达水平明显升高,与肝癌的发生发展及转移相关.

Objective To investigate the expression of phosphorylated protein p27 in hepatocellular carcinoma （HCC） tissue and its relationship to clinicopathological features of HCC.Methods The method Ti4＋ -IMAC combined with LC-MS/MS and MS/MS/MS （ LC-MS2-MS3 ） was used to detect phosphorylated protein in HCC tissue samples,and the expression of P-p27T187 and P-p27S10 was determined by Western blotting in the resected liver tumor tissues and corresponding adjacent tissues from 40 patients with HCC and normal liver tissues from 12 patients with benign lesions.Results The phosphorylated protein p27 was identified by the method Ti4＋ -IMAC combined with LC-MS2-MS3 from the HCC tissues,and also was the IPI number,phosphorylated sites,protein kinase and so on.The expression of P-p27T187 and P-p27S10 detected by Western blotting was significantly higher in HCC tissue （0.635 ±0.108） than in the corresponding adjacent tissues （0.306 ± 0.078 ） and normal liver tissue （ 0.149 ± 0.031 ） （ P 〈 0.05 ）.The expression of P-p27T187 and P-p27S10 was significantly associated with pathological grade,clinical stage and metastasis in HCC （ P 〈 0.05 ）,however not significantly correlated to other clinicopathological features （P 〉0.05）.Conclusion Phosphorylated protein p27 expression is increased in HCC tissue as compared with adjacent tissues and normal tissues,which may be related to the occurrence/development and metastasis in HCC.