丝氨酸肽酶抑制因子Kazal型1对肝癌细胞增殖的影响及其机制  

Effect of serine protease inhibitor Kazal-type 1 on the proliferation of hepatocellular carcinoma cell and its mechanism

作　　者：赵卫明[1] 李晓 余国营 王改平[1] 常翠芳[1] ZHAO Wei-ming;LI Xiao;YU Guo-ying;WANG Gai-ping;CHANG Cui-fang(Collage of Life Sciences,He’nan Normal University,State Key Laboratory for Cell Differentiation and Regulation,He’nan Xinxiang 453007,China)

机构地区：[1]河南师范大学生命科学学院,河南省-科技部共建细胞分化国家重点实验室培育基地,河南新乡453007

出　　处：《解剖学报》2023年第6期695-702,共8页Acta Anatomica Sinica

基　　金：河南省自然科学基金(212300410362);河南省高等学校重点科研项目(22A180006);河南省重大科技攻关项目(212102310879);肺纤维化生物学过程与防治高等学校学科创新引智基地(111基地)。

摘　　要：目的探讨丝氨酸肽酶抑制因子Kazal型1(SPINK1)对肝癌细胞RH-35增殖的影响及其分子机制。方法利用基因表达综合(GEO)数据库分析Spink1基因在肝癌患者和大鼠肝癌模型中的表达,用重组大鼠SPINK1蛋白(rrSPINK1)处理RH-35细胞,采用MTT、流式细胞术、5-乙炔基-2’-脱氧尿苷(EdU)等方法检测SPINK1对RH-35细胞增殖的影响,Real-time PCR和Western blotting检测SPINK1信号通路、细胞增殖和凋亡相关基因的表达。结果SPINK1在肝癌和大鼠肝癌模型中显著高表达;与对照组相比,rrSPINK1处理的RH-35细胞活力,EdU阳性细胞率,S期和G_2/M期细胞比例明显增加;而RH-35细胞凋亡无显著变化;rrSPINK1上调RH-35细胞中p38 MAPK和STAT信号通路相关基因/蛋白的表达;并且在肝癌中Spink1基因的表达与Mapk13、Stat1和Stat3基因正相关。结论SPINK1促进肝癌细胞中p38 MAPK和Janus激酶/信号转导及转录激活因子(JAK/STAT)的表达,促进肝癌细胞的增殖。Objective To explore the effect of serine protease inhibitor Kazal-type 1(SPINK1) on the proliferation of hepatocellular carcinoma cells RH-35 and its underling molecular mechanism. Methods Spink1 gene expression in liver cancer and rat liver cancer models were analyzed by Gene Expression Omnibus(GEO) data, RH-35 cells were treated with rrSPINK1 protein, the effect of rrSPINK1 on the proliferation and apoptosis of RH-35 cells was explored by MTT, 2'-deoxy-5-ethynyluridine(EdU) and flow cytometry, the molecular mechanism of SPINK1 regulating liver cancer were detected by Real-time PCR and Western blotting. Results The results showed that Spink1 gene was over-expressed significantly in liver cancer and rat liver cancer models, rrSPINK1-treated RH-35 cells showed increased viability, EdU-positive cell rate, and the proportion of cells in S phase and G_2/M phase compared to the control group. However, there was no significant difference in RH-35 cell apoptosis. rrSPINK1 up-regulated the expression of p38 MAPK and STAT pathway-related genes/proteins in RH-35 cells;and the expression of Spink1 gene in liver cancer was positively correlated with that of Mpak13, Stat1 and Stat3 genes. Conclusion SPINK1 may promote the expression of p38 MAPK and Januk kinase/signal transducers and activators of transcription(JAK/STAT), and promote the proliferation of hepatocellular carcinoma cells.

关 键 词：丝氨酸肽酶抑制因子Kazal型1 肝癌 细胞增殖 流式细胞术 人 大鼠 

分 类 号：Q28[生物学—细胞生物学]