PTEN、MMP-2在淋巴结侵袭性非小细胞肺癌细胞株H1299中的表达变化及其分子调控机制  

Expression changes and molecular regulation mechanisms of PTEN and MMP-2 in lymph node invasive non-small-cell lung cancer H1299 cells

作　　者：李璐 王凡 姚小艳 刘珍 LI Lu;WANG Fan;YAO Xiaoyan;LIU Zhen(Department of Clinical Laboratory,Shanghai First Maternity and Infant Hospital Affiliated to Tongji University,Shanghai 200092,China;不详)

机构地区：[1]同济大学附属第一妇婴保健院检验科,上海200092 [2]上海市浦东医院/复旦大学附属浦东医院检验科

出　　处：《山东医药》2022年第21期1-4,共4页Shandong Medical Journal

基　　金：复旦大学附属浦东医院院级人才项目(PX201703)。

摘　　要：目的观察同源性磷酸酶张力蛋白(PTEN)、基质金属蛋白酶2(MMP-2)在淋巴结侵袭性非小细胞肺癌(NSCLC)细胞株H1299中的表达变化,并探讨其分子调控机制。方法采用RT-qPCR法和Western Blotting法检测淋巴结侵袭性NSCLC细胞株H1299、淋巴结非侵袭性NSCLC细胞株A549中PTEN和MMP-2 mRNA及蛋白。采用慢病毒介导的脂质体转染技术建立过表达PTEN的H1299细胞,记为PTEN过表达组,以转染空载质粒的H1299细胞为对照,记为对照组,分别采用RT-qPCR法和Western Blotting法检测两组细胞中PTEN蛋白、MMP-2 mRNA及蛋白、Akt蛋白、p-Akt蛋白,采用荧光素酶报告基因实验检测转录因子NF-κb。结果H1299细胞中PTEN mRNA及蛋白的相对表达量均低于A549细胞,MMP-2 mRNA及蛋白的相对表达量均高于A549细胞(P均<0.01)。与对照组相比,PTEN过表达组细胞中PTEN蛋白相对表达量显著增高,MMP-2 mRNA及蛋白、p-Akt蛋白相对表达量显著降低(P均<0.01)。PTEN过表达组细胞中NF-κb活性低于对照组(P<0.01)。结论淋巴结侵袭性NSCLC细胞株H1299中PTEN表达降低、MMP-2表达升高,过表达PTEN可能是通过Akt/NF-κb信号通路抑制MMP-2的表达。Objective To investigate expression changes and molecular mechanism of phosphatase and tensin homology deleted on chromosome 10(PTEN)and matrix metalloproteinase-2(MMP-2)in lymph node invasive non-small-cell lung cancer(NSCLC)H1299 cells.Methods RT-qPCR and Western blotting were used to determine the mRNA and protein expression levels of PTEN and MMP-2 in NSCLC cells without lymph node metastasis(A549)and NSCLC cells with lymph node metastasis(H1299).Lentiviral-mediated lipofection technique was used to make the PTEN over-expressed in H1299 cells(the up-regulated PTEN group).H1299 cells which were transfected with vector plasmid were taken as the control group.The mRNA level of MMP-2 was detected by RT-qPCR and the protein expression levels of PTEN,MMP-2,Akt and p-Akt were detected by Western blotting.NF-κb was determined by using luciferase reporter gene experiment.Results Compared with A549 cells,the relative expression levels of PTEN mRNA and protein in H1299 cells decreased and the relative expression levels of MMP-2 mRNA and protein in H1299 cells increased(all P<0.01).Compared with the control group,the protein expression of PTEN increased,the mRNA and protein expression levels of MMP-2,and the protein expression of p-Akt decreased in the up-regulated PTEN group(all P<0.01).Meanwhile,the activity of NF-κB luciferase was lower in the up-regulated PTEN group than in the control group(P<0.01).Conclusions The relative expression of PTEN decreased and the relative expression of MMP-2 increased in lymph node invasive NSCLC H1299 cells.The over-expression of PTEN can inhibit the expression of MMP-2 by Akt/NF-κb signal pathway.

关 键 词：同源性磷酸酶张力蛋白 基质金属蛋白酶2 非小细胞肺癌 H1299细胞 

分 类 号：R734.2[医药卫生—肿瘤]