Chen Chao, Liang Qing-Yu, Chen Hui-Kang, Wu Pin-Fei, Feng Zhen-Yu, Ma Xiao-Ming, Wu Hao-Rong, Zhou Guo-Qiang
Department of General Surgery, Changshu Second People's Hospital, The Fifth Hospital Affiliated to Yangzhou University, Changshu, Jiangsu 215500, P.R. China.
Department of General Surgery, The First Hospital of Zhang Jia Gang, Zhangjiagang, Jiangsu 215600, P.R. China.
Oncol Lett. 2018 Aug;16(2):2427-2433. doi: 10.3892/ol.2018.8937. Epub 2018 Jun 8.
DNA-damage regulated autophagy modulator 1 (DRAM1) is known as a target of TP53-mediated autophagy, and has been reported to promote the migration and invasion abilities of glioblastoma stem cells. However, the precise contribution of DRAM1 to cancer cell invasion and migration, and the underlying mechanisms remain unclear. In the present study, small interfering (si)RNA or short hairpin RNA mediated knockdown of DRAM1 was performed in hepatoblastoma cells and the migration and invasion abilities were detected and . To investigate the underlying mechanisms, western blotting and immunofluorescence were used to detect the expression of autophagy-associated proteins and epithelial-mesenchymal-transition (EMT)-associated markers. The results showed that DRAM1 knockdown by specific siRNA abrogated cell autophagy, as well as inhibited the migration and invasion of HepG2 cells in Transwell assays, which may be reversed by rapamycin treatment. In addition, DRAM1 knockdown increased the expression of E-Cadherin while decreased the expression of vimentin in HepG2 cells, which was also be reversed by rapamycin treatment. Taken together, these results suggest that DRAM1 is involved in the regulation of the migration and invasion of HepG2 cells via autophagy-EMT pathway.
DNA损伤调节自噬调节剂1(DRAM1)是TP53介导的自噬的一个靶点,据报道它能促进胶质母细胞瘤干细胞的迁移和侵袭能力。然而,DRAM1对癌细胞侵袭和迁移的确切作用及其潜在机制仍不清楚。在本研究中,在肝母细胞瘤细胞中通过小干扰(si)RNA或短发夹RNA介导敲低DRAM1,并检测其迁移和侵袭能力。为了探究潜在机制,采用蛋白质免疫印迹法和免疫荧光法检测自噬相关蛋白和上皮-间质转化(EMT)相关标志物的表达。结果显示,特异性siRNA敲低DRAM1可消除细胞自噬,并且在Transwell实验中抑制HepG2细胞的迁移和侵袭,而雷帕霉素处理可逆转这种抑制作用。此外,敲低DRAM1可增加HepG2细胞中E-钙黏蛋白的表达,同时降低波形蛋白的表达,雷帕霉素处理也可逆转这种变化。综上所述,这些结果表明DRAM1通过自噬-EMT途径参与调节HepG2细胞的迁移和侵袭。