溶血磷脂酸(LPA)对肝癌细胞的影响及相关机制的初步探讨  

Effect of lysophosphatidic acid on hepatoma cells and related mechanism

作　　者：赵燕颖 韩振琦[1] 邹艳平[1] 李云鹏 徐涛 刘丽艳 程海涛 ZHAO Yanying;HAN Zhenqi;ZOU Yanping;LI Yunpeng;XU Tao;LIU Liyan;CHENG Haitao(Department of Gastrointestinal Endoscopy,The Affiliated Hospital of Changchun University of Traditional Chinese Medicine,Changchun 130021,China;Department of Gastroenterology,The FAW General Hospital of Jilin Province,Changchun 130011,China)

机构地区：[1]长春中医药大学附属医院消化内镜科,长春130021 [2]吉林省一汽总医院消化内科,长春130011

出　　处：《临床肝胆病杂志》2023年第11期2623-2628,共6页Journal of Clinical Hepatology

基　　金：吉林省卫健委课题(2021LC093)。

摘　　要：目的探讨溶血磷脂酸(LPA)在肝癌患者中的表达情况和对肝癌恶性生物学行为的影响和调控机制。方法收集2016年1月—2022年12月吉林省一汽总医院收治的26例肝癌患者,28例肝硬化患者以及28例体检人群作为对照。用ELISIA方法检测伴有腹腔积液的肝癌和肝硬化患者血浆及腹腔积液中LPA含量,同时检测正常人群血浆中LPA含量,明确LPA在肝癌和肝硬化患者等不同人群中的表达差异。MTT细胞增殖实验和细胞迁移实验检测LPA和其抑制剂-百日咳毒素(PTX)对SMMC7721细胞增殖、迁移和侵袭能力的影响。为探讨LPA和其受体结合后对Rho A及其上下游的FAK、P53表达的影响,本研究采用q PCR和Western Blot法检测LPA对SMMC7721细胞P53、FAK、Rho A的m RNA和蛋白表达水平的影响。计量资料多组间样本均数比较采用单因素方差分析,两两比较采用SNK-q检验。结果肝癌患者血浆LPA浓度(4.99±0.55)μmol/L和腹腔积液中LPA浓度(5.19±0.63)μmol/L明显高于肝硬化患者(2.63±0.43)μmol/L、(2.91±0.46)μmol/L,肝癌患者血浆LPA亦明显高于正常人群(1.61±0.39)μmol/L,差异均有统计学意义(P值均<0.05)。细胞增殖试验显示LPA可明显促进SMMC7721细胞增殖,细胞增殖率随剂量和时间增加而增加,尤其中高剂量组增殖率明显高于对照组,差异具有统计学意义(P<0.05)。而PTX抑制其增殖能力,且具有时间依赖性,各组差异具有统计学意义(P<0.05)。72 h高剂量LPA的增殖率是对照组的3.6倍,PTX组是对照组的0.6倍,二者联合组是对照组的1.2倍。另外,LPA可增加肝癌细胞的迁袭能力,而PTX可抑制其迁移,随时间增加上述能力增加,各组差异具有统计学意义(P<0.05)。72 h高剂量LPA的迁袭率是对照组的3.09倍,PTX组是对照组的0.4倍,二者联合组是对照组的0.99倍。qPCR和Western Blot结果显示LPA处理后的SMMC7721细胞P53的mRNA和蛋白表达水平明显减少,而FAK、RhoA的mRNA和蛋白表达水平�Objective To investigate the expression of lysophosphatidic acid(LPA)in patients with liver cancer,as well as its influence on malignant biological behavior of liver cancer and related regulatory mechanism.Methods From January 2016 to December 2022,26 patients with liver cancer,28 patients with liver cirrhosis,and 28 individuals undergoing physical examination were enrolled.ELISIA was used to measure the content of LPA in plasma and peritoneal effusion of the patients with liver cancer or liver cirrhosis accompanied by peritoneal effusion,and the content of LPA was measured in plasma of the normal population at the same time,so as to clarify the difference in the expression of LPA in different populations,such as the patients with liver cancer and those with liver cirrhosis.MTT cell proliferation assay and cell migration assay were used to observe the influence of LPA and its inhibitor pertussis toxin(PTX)on the proliferation,migration,and invasion of SMMC7721 cells.In order to investigate the effect of LPA on the expression of RhoA and its upstream and downstream molecules FAK and P53 after binding to its receptor,qPCR and Western blot were used to observe the effect of LPA on the mRNA and protein expression levels of P53,FAK,and RhoA in SMMC7721 cells.A one-way analysis of variance was used for comparison of the means of continuous data between multiple groups,and the SNK-q test was used for comparison between two groups.Results Compared with the patients with liver cirrhosis,the patients with liver cancer had a significantly higher concentration of LPA in plasma(4.99±0.55μmol/L vs 2.63±0.43μmol/L,P<0.05)and peritoneal effusion(5.19±0.63μmol/L vs 2.91±0.46μmol/L,P<0.05),and the patients with liver cancer also had a significantly higher level of plasma LPA than the normal population(4.99±0.55μmol/L vs 1.61±0.39μmol/L,P<0.05).The cell proliferation assay showed that LPA significantly promoted the proliferation of SMMC7721 cells,and cell proliferation rate increased with the increase in dose and time;i

关 键 词：肝肿瘤 溶血磷脂素类 百白咳毒素 

分 类 号：R735.7[医药卫生—肿瘤]