摘要
背景与目的:Fibulin-5在肺癌组织中低表达,具有抑癌作用。高迁移率族蛋白B1(high mobility group box 1,HMGB1)在肺癌中高表达,能够促进肿瘤的侵袭转移。该研究旨在探讨Fibulin-5抑制肺癌细胞增殖和转移的分子机制。方法:该研究首先检测了肺上皮细胞和肺癌细胞中Fibulin-5和HMGB1的表达,然后利用转染试剂将Fibulin-5过表达质粒和HMGB1的si RNA转染人A549细胞。实现Fibulin-5过表达和HMGB1低表达后,采用MTT实验检测细胞增殖情况,Transwell实验检测细胞的侵袭和迁移能力。本研究采用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)检测A549细胞中HMGB1 m RNA表达变化,采用酶联免疫吸附剂测定实验(enzyme-linked immunosorbent assay,ELisa)检测HMGB1蛋白的分泌;采用蛋白[质]印迹法(Western blot)检测HMGB1、cyclin D1、基质金属蛋白酶(matrix metalloproteinases,MMPs)和TLR4/NF-κB通路相关蛋白的表达变化。结果:在肺癌细胞A549中,Fibulin-5低表达,HMGB1高表达。过表达Fibulin-5和低表达HMGB1后,HMGB1、cyclin D1、MMP2、MMP7和MMP9的表达均明显降低,A549细胞的增殖、侵袭和迁移能力明显减弱(P<0.05);此外,过表达Fibulin-5下调了TLR4、MyD88、p-p65的表达,上调了IκBα的表达(P<0.05)。结论:Fibulin-5可能是通过抑制HMGB1的表达以及其下游的TLR4/NF-κB通路,抑制肺癌细胞的增殖、侵袭和迁移的过程。
Background and purpose： The expression of Fibulin-5, a tumor suppressor, is reduced in lung cancer tissues. High mobility group box 1 （HMGB1）, overexpressed in lung cancer, can promote the invasion and metastasis of the tumor. This study aimed to investigate the molecular mechanism of Fibulin-5 inhibiting the proliferation and metastasis of lung cancer cells. Method： Firstly the expressions of Fibulin-5 and HMGB1 in lung epithelial cells and lung cancer cells were detected. Then transfection reagents were used to transfect Fibulin-5 plasmid and HMGB1 siRNA into A549 cells, which achieved Fibulin-5 overexpression and HMGB1 down-regulation. MTT was used to detect the cell proliferation. Transwell was used to examine the ability of cell invasion and migration. HMGB1 mRNA expression was detected by real-time fluorescence quantitative polymerase chain reaction （RTFQ-PCR）. The secretion of HMGB 1 protein was analyzed by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the levels of HMGB 1, cyclin D 1, matrix metalloproteinases （MMPs）, and TLR4/NF-KB pathway related proteins. Results： In A549 cell, Fibulin-5 expression was down-regulated and HMGB1 expression was up-regulated. After overexpression of Fibulin-5 or knockdown of HMGB 1 in A549 cells, the levels of HMGB 1, cyclin D1, MMP2, MMP7 and MMP9 were significantly reduced; the abilities of lung cancer cell proliferation, invasion and migration werenotably suppressed （P〈0.05）. In addition, Fibulin-5 overexpression decreased the protein expressions of TLR4, MyD88, p-p65, increased the expression of IkBet （P〈0.05）. Conelusion： Fibulin-5 may inhibit lung cancer cell proliferation, invasion and migration via inhibiting the expression of HMGB 1 and its downstream TLR4/NF-rd3 pathway.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2017年第10期761-769,共9页
China Oncology
