摘要
目的:探讨沉默Toll样受体4(TLR4)基因后对乳腺癌细胞侵袭性的影响,为乳腺癌治疗提供新思路。方法:(1)用重组慢病毒感染乳腺癌MD–MBA–231细胞,干扰TLR4基因的表达,Western blot法检测MDA–MB–231/shTLR4(沉默)的TLR4蛋白表达水平。(2)加入外源性刺激因子高迁移率族蛋白1(HMGB1)并设置空白对照磷酸盐缓冲液(PBS),Western blot法检测在乳腺癌细胞MDA–MB–231/shTLR4(沉默)及MDA–MB–231/shControl(对照)细胞中下游信号因子相关蛋白核因子κB(NF–κB)、白细胞介素(IL)–6、血管内皮生长因子a(VEGFa)的表达水平。结果:(1)在沉默TLR4基因后,MDA–MB–231/shTLR4细胞的TLR4蛋白水平均明显降低,差异具有统计学意义(P<0.05)。(2)沉默TLR4基因后,HMGB1对于MDA–MB–231/shTLR4(沉默)细胞中下游信号因子相关蛋白NF–κB、IL–6、VEGFa的表达水平促进作用较MDA–MB–231/sh Control(对照)明显降低,差异具有统计学意义(P<0.01)。结论:沉默TLR4基因后,MDA–MB–231/sh TLR4(沉默)下游信号因子相关蛋白NF–κB、IL–6、VEGFa表达明显降低,从而降低了乳腺癌细胞的侵袭性。
Objective To investigate the effect of the toll-like receptor 4（TLR4） silencing on the invasion of breast cancer cells and therefore pave the way for developing a novel breast cancer treatment strategy. Method（1） Recombinant lentiviruses were used to infect MD-MBA-231 cell lines to interfere the expression of TLR4. The expression of TLR4 was examined in MDA–MB–231/sh TLR4 via Western Blot.（2） Exogenous high mobility group box-1（HMGB1） was used to stimulate the TLR4-silenced MDAMB-231 cells（MDA-MB-231/sh TLR4） and TLR-4 expressing MDA-MB-231 cells（MDA-MB-231/sh Control）, where phosphate buffer saline（PBS） was used as negative control. The expressions of downstream signaling factor were examined, including nudear factor κB（NF-κB）, IL-6 and VEGFa. Result（1） The expression of TLR4 significantly decreased in TLR4-silenced MDA-MB-231 cells（P〈0.05）.（2） The enhancing effect of HMGB1 on the expression of NF-κB, IL-6 and VEGFa was significantly reduced in MDA-MB-231/sh TLR4, compared with MDA-MB-231/sh Control.（P〈0.01）. Conclusion After silence TLR4 gene, the expression of NF-κB, IL-6 and VEGFa downstream of MDA-MB-231/sh TLR4（silenced） signalling was significantly reduced, which reduced the invasiveness of breast cancer cells.
作者
吴坤琳
张欣
陈祥锦
WU Kun-lin;ZHANG Xin;CHEN Xiang-jin(The First Affiliated Hospital of Fujian Medical University, Fujian Fuzhou 35000)
出处
《深圳中西医结合杂志》
2018年第4期1-3,F0003,共4页
Shenzhen Journal of Integrated Traditional Chinese and Western Medicine
基金
福建省自然科学基金项目资助课题(2015J01386)
福建省卫生教育联合攻关计划项目资助课题(WKJ2016-2-26)
