Notch3 pathway mediates SAHA-induced apoptosis in human small-cell lung cancer H446 cells
10.3969/j.issn.1000-4718.2016.09.004
- VernacularTitle:Notch3信号通路介导 SAHA 诱导的小细胞肺癌H446细胞凋亡
- Author:
Honglian CHEN
;
Hui LIU
;
Xuguang YANG
;
Lei YUAN
- Publication Type:Journal Article
- Keywords:
Suberoylanilide hydroxamic acid;
Small-cell lung cancer;
Notch3;
Apoptosis;
H446 cells
- From:
Chinese Journal of Pathophysiology
2016;32(9):1556-1561
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the effect of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hu-man small-cell lung cancer H446 cells and its possible mechanism.METHODS: H446 cells were incubated in the medi-um containing SAHA.CCK-8 assay was used to detect the anti-tumor effect of SAHA on the H446 cells, and IC50 values of SAHA were calculated.Flow cytometry was used to analyze the apoptosis.After Notch3 gene was silenced, the pro-apopto-tic effect of SAHA on the H446 cells was inhibited ( P <0.05).Eukaryotic expression plasmid containing N3ICD was transfected into the H446 cells, so that N3ICD was expressed in the H446 cells.The mRNA expression of Notch3 was measured by RT-PCR.The protein levels of Notch3, N3ICD, Puma and cleaved caspase-3 were determined by Western blot.RESULTS: SAHA remarkably reduced the cell viability in a dose-dependent manner (P <0.05), and the IC50 value of SAHA was 1.91 μmol/L.SAHA induced apoptosis in a dose-dependent manner (P <0.05).The expression of Notch3 gene was negative in the H446 cells, SAHA reactivated Notch3 gene and Notch3 pathway in a dose-dependent manner (P <0.05).Notch3 knockdown inhibited apoptosis induced by SAHA (P <0.05).Over-expression of N3ICD up-regula-ted the protein levels of Puma and cleaved caspase-3.CONCLUSION: SAHA induces apoptosis in human small-cell lung cancer H446 cells by activating Notch3 pathway and up-regulating the protein level of Puma.
