摘要
目的分析PBLD对HepG2细胞增殖、侵袭及转移的影响,并初步探讨其发挥作用的分子机制。方法首先利用细胞转染技术构建PBLD过表达的HepG2稳转细胞株,CCK-8法检测PBLD过表达后肝癌细胞增殖能力的变化,Transwell实验检测PBLD过表达后HepG2细胞侵袭、转移能力的变化,最后利用Western-blot技术筛选并验证PBLD可能的下游信号通路。结果 PBLD过表达后HepG2细胞的增殖能力被显著抑制(P<0.05),同时,发生侵袭、迁移的细胞数量与空载对照细胞相比显著减低[(68±12.5)vs.(19±10.21),P<0.01;(30±8.83)vs.(11±4.41),P<0.01]。进一步Western-blot检测结果显示,PBLD过表达可显著抑制MAPK信号通路相关分子的表达。结论 PBLD可通过MAPK信号通路抑制Hep G2细胞的增殖、侵袭和转移。
Objective To investigate the effects of PBLD on the proliferation, migration and invasion of HepG2 cells and to explore the underlying molecular mechanism. Methods Firstly, we constructed PBLD overexpression cell line using liposome transfection technique. CCK-8 proliferation assay was performed to examine the effects of PBLD on cell growth in vitro. Transwell migration and invasion assay was carried out to investigate the effects of PBLD on the migration and invasion of HepG2 cells. Finally, the downstream genes mediating the effects of PBLD in HepG2 cells were identified by Western-blot examination. Results CCK-8 proliferation assay showed that upregulation of PBLD significantly inhibited the proliferation rate of HepG2 cells （P 〈 0.05）. Transwell migration and invasion assay showed that HepG2 cells migrated slower and had less ability to invade through the Matrigel-coated inserts when PBLD was upregulated [（68 ± 12.5） vs. （19 ± 10.21）,P 〈 0.01; （30 ± 8.83） vs. （11 ± 4.41）, P〈0.01]. The expressions ofgenesin MAPK signal pathwaywere significantly inhibited in PBLD overexpression cells. Conclusion PBLD inhibited the growth, migration and invasion of HCC cells in vitro via MAPK signal pathway.
出处
《现代消化及介入诊疗》
2016年第4期523-526,共4页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
国家自然科学基金青年项目(81402028)
广东省自然科学基金(2015A030310082)
广东省海外合作项目(2014A050503041)
