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Δ40p53 对p53 促肿瘤细胞凋亡作用的影响.

Authors :
王贲士
赵红伟
乔录新
单军奇
侯庆生
陈德喜
郭洪亮
Source :
Chinese Journal of Oncology. 5/23/2017, Vol. 39 Issue 5, p332-338. 7p.
Publication Year :
2017

Abstract

Objective To investigate the effect of Δ40p53, an alternative spliced isoform of p53 lacking the N⁃ter minus, on the pro⁃apoptotic function of p53. Methods The wild⁃type p53 was ectopically expressed in HCT116⁃p53-/ -(endogenous Δ40p53 expression), HCT116⁃p53+/ +(wild⁃type p53) and H1299 (p53⁃null) cells by adenoviral delivery, while Δ40p53 plasmid were transfected into these cells to overexpress Δ40p53. The levels of Δ40p53 and p53 mRNA were detected by reverse transcription⁃polymerase chain reaction (RT⁃PCR) and quantitative PCR. The expression of related proteins was deter mined by Western blotting. The interaction of p53 and Δ40p53 was observed by co⁃immunoprecipitation assay. Calcein⁃AM/ propidium iodide (PI) staining and flow cytometry were used to detect the apoptotic rate of tested cells in each group. Results HCT116⁃p53-/ - cells expressed endogenous Δ40p53 isoform. Neither transcription nor protein expression of wild⁃type p53 was interfered by the increased expression of Δ40p53. Full length p53 and Δ40p53 could bind to each other. Calcein⁃AM/ PI staining showed that the apoptotic rates of H1299⁃Control, HCT116⁃p53-/ - ⁃Control, H1299+p53, HCT116⁃p53-/ - +p53, H1299+oxaliplatin (Oxa), HCT116⁃p53-/ - +Oxa, H1299+p53+Oxa and HCT116⁃p53-/ - +p53+Oxa groups were (2.50±0.47)%, (2.40±0.32)%, (5.20±0.58)%,(4.10±0.18)%, (22.40±1.73)%, (19.30±1.11)%, (29.90±1.15)% and (39.30±2.26)%, respectively. It was statistically significant between H1299+p53+Oxa and HCT116⁃p53-/ - + p53+Oxa groups (t = 3.721, P = 0.0205). Moreover, the apoptotic rates of H1299⁃Control, H1299+Δ40p53, H1299+p53, H1299+p53+Δ40p53, H1299+Oxa, H1299+Δ40p53+Oxa, H1299+p53+Oxa and H1299+p53+Δ40p53+Oxa groups were (2.60±0.35)%, (2.20±0.17)%, (4.80±0.49)%, (4.90±1.10)%, (20.30±1.10)%, (19.60±1.45)%, (27.90±1.39)%, (35.20±1.43)%, respectively. Furthermore, flow cytometry assay showed that the apoptotic rates of above cells were (2.70± 0.32)%, (2.20± 0.24)%, (4.60±0.48)%, (3.90±0.67)%, (19.30±1.11)%, (17.70±0.66)%, (28.30±2.76)% and (37.50± 1.51)%, respectively. H1299+p53+Δ40p53+Oxa cells showed higher cell apoptosis than H1299+p53+Oxa cells (t =2.930, P = 0.042). Conclusion Δ40p53 isoform can bind to full⁃length p53, and enhance its pro⁃apoptotic function in tumor cells. [ABSTRACT FROM AUTHOR]

Details

Language :
Chinese
ISSN :
02533766
Volume :
39
Issue :
5
Database :
Academic Search Index
Journal :
Chinese Journal of Oncology
Publication Type :
Academic Journal
Accession number :
123184612
Full Text :
https://doi.org/10.3760/cma.j.issn.0253⁃3766.2017.05.003