Effect of rapamycin on the biological behaviors of human non-Hodgkin lymphoma Raji cells and its mechanism
10.3760/cma.j.issn.1009-9921.2018.02.007
- VernacularTitle:雷帕霉素对人非霍奇金淋巴瘤Raji细胞生物学行为的影响及其机制研究
- Author:
Hua LI
1
;
Yun ZHOU
;
Xuechao LIU
;
Yuan TANG
;
Xingchu WEN
;
Yang WANG
Author Information
1. 071000,河北省保定市第一医院肿瘤外科
- Keywords:
Lymphoma,non-Hodgkin;
Sirolimus;
Apoptosis;
Caspase 3;
Caspase 9;
Genes,bcl-2;
Genes;
p53
- From:
Journal of Leukemia & Lymphoma
2018;27(2):89-94
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of rapamycin on the biological behaviors of human non-Hodgkin lymphoma Raji cells with different concentrations and time, and to explore its mechanism. Methods Different concentrations (0, 10, 50, 100, 250, 500 nmol/L) of rapamycin were used in Raji cells for 24, 48, 72 h respectively. The apoptosis of Raji cells was detected by using CCK-8 method, and flow cytometry was used to detect the cell apoptosis and cycle of Raji cells. The enzymatic activity of Caspase-3 and Caspase-9 in Raji cells was detected by Caspase-3 and Caspase-9 activity testing kit. The expressions of bcl-2, p53 protein and mRNA were detected by Western blot method and reverse transcription-polymerase chain reaction (RT-PCR). Results The proliferative inhibition rate of Raji cells was increased from (23.7 ± 4.2)％to (51.7±3.7)％, the cell apoptosis rate was increased from (4.9±1.9)％to (20.5±1.5)％, the proportion of G0/G1 was increased from (40.8±1.4) ％to (63.6±1.7) ％, the Caspase-3 enzyme activity of Raji cell in 24 h was increased from 0.16±0.05 to 1.08±0.04, Caspase-9 enzyme activity was increased from 0.19±0.04 to 1.34± 0.06 after 24 h with the increasing concentration of rapamycin from 0 nmol/L to 500 nmol/L (P<0.01). The mRNA of bcl-2 was decreased from 0.90±0.03 to 0.46±0.03, and mRNA of p53 was increased from 2.51±0.41 to 5.85±0.21. The protein expression of bcl-2 was reduced and the protein expression of p53 was increased. The experimental results of Raji cells in 48 h and 72 h were consistent with the experimental results of 24 h. Conclusion Rapamycin may inhibit the proliferation of Raji cells through Caspase-3, Caspase-9, bcl-2, p53 and induce its cell apoptosis.
