Involvement of Akt in synergistic effects of thermo-chemotherapy in human small cell lung cancer H446 cell apoptosis
- VernacularTitle:Akt通路在热化疗联合诱导人小细胞肺癌细胞凋亡中的作用
- Author:
Wang LIN
1
;
Liu XIN-KUI
;
Shi XIU-QIN
Author Information
1. 郑州大学附属第一医院
- Keywords:
Akt pathway;
thermo-therapy;
lung tumor;
H446 cell;
apoptosis
- From:Chinese Journal of Cancer
2010;29(4):429-433
- CountryChina
- Language:Chinese
-
Abstract:
Background and Objective: Akt pathway plays an important role in cell growth and apoptosis.This study was to characterize the role of Akt in the synergistic effects of thermo-chemotherapy on lung cancer cell growht and its underlying mechanisms.Methods: H446 cells were subjected to different thermo-chemotherapy schemes: 43℃+paclitaxel(120 μg/L)(thermo-chemotherapy group),43℃+paclitaxel(120 μg/L)+Wortmannin(1 μmol/L,PI3K/Akt pathway inhibitor)(Wortmannin group),43℃+paclitaxel(120 μg/L)+N-acety-L-cysteine(NAC)(30 μmol/L,reactive oxygen species,ROS inhibitor)(NAC group),and paclitaxel(120 μg/L)group.The cells without any treatment were used as the control.MTT assay was conducted to measure the cell proliferation rate.Cell apoptosis was analyzed by flow cytometry(FCM).ROS was detected with fluorescence.Phosphorylation of Akt and the expressions of Caspase-3 were determined by Western blot.Results: The cell proliferation rate was significantly lower in the thermo-chemotherapy group than in the control and the chemotherapy groups[(59.83±3.36)% vs.(100.00±0.00)% and(69.16±2.95)%,P<0.05].The rate of cell apoptosis was the highest in the thermo-chemotherapy group(27.59±5.47)%(P<0.05).The ROS expression level was higher in the cells of thermo-chemotherapy group(102.14±18.34)than in the other groups(P<0.05),which could be inhibited by NAC(28.01±1.19),but not by the PI3K inhibitor Wortmannin(99.87±8.35).Phosphorylation of Akt significantly decreased in the thermo-chemotherapy group(0.69±0,03)(P<0.05),which could be blocked by Wortmannin(0.00±0.00),but increased by NAC(1.05±0.29)(P<0.05).The expression level of Caspase-3 was higher in the thermo-chemotherapy group(1.07±0.08)than in other groups(P<0.05).Conclusion: Thermo-chemotherapy has a stronger inhibitory effect than chemotherapy alone in lung tumor cell growth,probably through induction of ROS production and subsequent inhibition of Akt pathway activation and Caspase pathway-induced cancer cell apoptosis.
