Identification of role of PI3K in mediating necroptosis of L929 cells induced by tumor necrosis factor alpha
10.7644/j.issn.1674-9960.2017.01.007
- VernacularTitle:PI3 K在肿瘤坏死因子诱导L929细胞程序性坏死过程中的调控作用研究
- Author:
Xixi CHANG
;
Shiping HU
;
Yu WANG
;
Lili WANG
;
Shuai WU
;
Zicheng WANG
;
Zhiyan DU
;
Jiyun YU
;
Yi ZHANG
;
Guozhu CHEN
- Keywords:
phosphatidylinositol-3-kinase;
tumor necrosis factor alpha;
necroptosis;
protein kinase B;
MLKL
- From:Military Medical Sciences
2017;41(1):25-32
- CountryChina
- Language:Chinese
-
Abstract:
Objective To identify the role of phosphatidylinositol-3-kinase(PI3K) in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) and the involved mechanism.Methods Knockdown of p110α,receptor-interacting protein 1(RIP1) or both p110αand RIP1 was mediated by the specific short hairpin RNA (shRNA) lentivirus and verified by RT-PCR or Western blotting .In addition , Western blotting was used to detect phosphorylation of mixed lineage kinase domain-like protein(MLKL) and protein kinase B(AKT) or tetramerization of MLKL.Cell death was measured by micros-copy and flow cytometry.Results AKT phosphorylation and TNFα-induced necroptosis of L929 cells were suppressed by the inhibitors of PI3K or AKT, as well as p110αknockdown.Moreover, RIP1 knockdown did not inhibit L929 cell death induced by TNFαplus Z-VAD, but the RIP1-independent necroptosis was inhibited by p 110αknockdown.In addition, p110αknockdown suppressed MLKL phosphorylation and tetramerization induced by TNFαwith Z-VAD in L929 cells. Conclusion PI3K mediates necroptosis of L929 cells induced by TNFαby activating AKT and MLKL, respectively.
