caa a22 4500 606234152 CHVBK 20210128101235.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s11010-014-2253-6 doi (NATIONALLICENCE)springer-10.1007/s11010-014-2253-6 DNA-PKcs deficiency sensitizes the human hepatoma HepG2 cells to cisplatin and 5-fluorouracil through suppression of the PI3K/Akt/NF- κ B pathway [Elektronische Daten] [Yuan Fang, Zongtao Chai, Dansong Wang, Tiantao Kuang, Wenchuan Wu, Wenhui Lou] The aim of the present study was to investigate the effects of DNA-PKcs deficiency on the chemosensitivity of human hepatoma HepG2 cells to cisplatin (CDDP) and 5-fluorouracil (5-Fu), and to explore the underlying molecular mechanism. After transfection with DNA-PKcs siRNA or control siRNA, HepG2 cells were exposed to combination treatment of CDDP and 5-Fu. The cell viability, DNA damage, cell apoptosis, intracellular reactive oxygen species and glutathione (GSH) level, expression of apoptosis related proteins, activity of phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway, and nuclear factor-κB (NF-κB) pathways were assessed. The combination of CDDP and 5-Fu had a synergistic cytotoxic effect in HepG2 cells in terms of the cell viability, DNA damage, apoptosis, and oxidative stress level. DNA-PKcs siRNA could sensitize the HepG2 cells to the combined treatment. DNA-PKcs suppression further reduced the Akt phosphorylation level and Bcl-2 expression in HepG2 cells exposed to CDDP and 5-Fu, but enhanced the expression of pro-apoptotic proteins p53 and caspase-3. Moreover, CDDP could inhibit the transcriptional activity of NF-κB through degradation of IkB-α, while 5-Fu alone seemed in some extent increases the NF-κB activity. The combined treatment with CDDP and 5-Fu resulted in significantly decrease of the transcriptional activity of NF-κB, which was further aggravated by DNA-PKcs siRNA treatment. In conclusion, DNA-PKcs suppression had complementary effects in combination with CDDP and 5-Fu treatment in HepG2 cells, which was associated with suppression of NF-κB signaling pathway cascade, activation of caspase-3 and p53, as well as down-regulation of Bcl-2 and GSH. Springer Science+Business Media New York, 2014 DNA-PKcs nationallicence HepG2 cells nationallicence Apoptosis nationallicence Oxidative stress nationallicence NF- κ B nationallicence PI3K/AKT pathway nationallicence Bcl-2 nationallicence GSH nationallicence Fang Yuan Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut Chai Zongtao Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital, Liver Cancer Institute, Fudan University, 200032, Shanghai, People's Republic of China aut Wang Dansong Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut Kuang Tiantao Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut Wu Wenchuan Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut Lou Wenhui Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 399/1-2(2015-01-01), 269-278 0300-8177 399:1-2<269 2015 399 11010 https://doi.org/10.1007/s11010-014-2253-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-014-2253-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Fang Yuan Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Chai Zongtao Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Zhongshan Hospital, Liver Cancer Institute, Fudan University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Wang Dansong Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Kuang Tiantao Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Wu Wenchuan Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Lou Wenhui Department of General Surgery, Zhongshan Hospital, Fudan University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 399/1-2(2015-01-01), 269-278 0300-8177 399:1-2<269 2015 399 11010