1.Mechanism of BCL2L2-PABPN1 expression induced by sodium arsenite and its metabolites in 16HBE cells
SHI Ya YIN Jin yao WU Jiang JIANG Cheng lan ZHAO Rui huan ZHOU Qian HE Yue feng
China Occupational Medicine 2022;49(05):522-
Objective - - (BCL2L2)- ( )
To investigate the differential expression of the fusion gene BCL 2 like protein 2 poly A
(PABPN1) ( )
binding protein nuclear 1 induced by sodium arsenite SA and its methylated metabolites in 16HBE cells and the
Methods ) ,
related mechanism. i The 16HBE cells exposed to SA at concentrations of 1.5 3.0 and 4.5 µmol/L were set as
-, - -
low medium and high dose arsenic exposure groups. The 16HBE cells exposed to 4.5 µmol/L monomethylarsonic acid
( ), ( ) ,
MMA dimethylarsonic acid DMA and SA were set as MMA group DMA group and SA group. The 16HBE cells without
, BCL2L2-PABPN1
toxic stimulation were set as control group. After the cells were cultured for 48 hours the expression of was
- ( - ) ) ( )
detected by quantitative real time polymerase chain reaction qRT PCR . ii Two small interfering RNA siRNA silencing
基金项目:国家自然科学基金( ); 年云南省科技厅昆明医科大学应用基础研究联合专项面上项目
82160607 2021
( )
202101AY070001-054
作者简介:施雅( —),女,在读大学本科生,主要从事劳动卫生与环境卫生学研究;尹锦瑶( —),女,在读劳动卫生与环境卫
2001 1995
生学硕士研究生,主要从事劳动卫生与环境卫生学研究;施雅和尹锦瑶为共同第一作者
通讯作者:何越峰教授,博士研究生导师,- :
E mail heyuefeng@kmmu.edu.cn中国职业医学 年 月第 卷第 期 , , , · ·
2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 523
BCL2L2-PABPN1, -
fragments were designed and transfected into 16HBE cells to knockdown which were set as siRNA 1 group
- - BCL2L2-PABPN1
and siRNA 2 group. Non transfected control group without knockdown of transfection was set up. After
, BCL2L2-PABPN1 -
culturing for 48 hours the expression level of in the three groups of cells was detected by qRT PCR. The cell
-
survival rate and early apoptosis rate were detected by MTS method and JC 1 mitochondrial membrane potential detection
, ( ) ,
method respectively. The apoptosis was detected by Hoechest33342/propidium iodide PI double staining and the expression
- Results )
level of P53 signaling pathway related proteins was detected by Western blotting. i The relative expression of
BCL2L2-PABPN1 (P ) BCL2L2-
in 16HBE cells increased with the increasing SA doses <0.01 . The relative expression of
PABPN1 - , - -
in high dose arsenic exposure was higher than that in control group low dose and medium dose arsenic exposure
( P ) BCL2L2-PABPN1 ,
groups all <0.05 . The relative expression of in SA group was higher than those in control group MMA
( P ) BCL2L2-PABPN1
group and DMA group all <0.05 . The relative expression of showed no significant difference between
, ( P ) ) BCL2L2-PABPN1
control group MMA group and DMA group all >0.05 . ii The relative expression levels of and cell
- - - ( P )
survival rate in siRNA 1 group and siRNA 2 group were lower than those in non transfected control group all <0.05 .
, (P )
However there was no significant difference in the early apoptosis rate among the three groups >0.05 . The results of
-
Hoechest33342/PI double staining showed that the number of nuclear shrinkage and early apoptotic cells in siRNA 1 group and
- - , -
siRNA 2 group was higher than that in non transfected control group. The relative protein expression levels of P53 phospho
, - - , - - ( P )
p53 BCL 2 associated death promoter P21 and cytochrome C in siRNA 1 group and siRNA 2 group were higher all <0.05 ,
- - P
and the relative protein expression levels of P53 up regulated modulator of apoptosis were lower (all <0.05), when compared
- Conclusion
with the non transfected control group. SA may block the apoptosis of 16HBE cells by inducing the expression of
BCL2L2-PABPN1
fusion gene . The mechanism may be related to the activation of P53 signaling pathway. The SA methylated
BCL2L2-PABPN1 BCL2L2-PABPN1 -
metabolites MMD and DMA had no effect on the expression of . may affect anti apoptosis
BCL2L2 PABPN1
through affecting the synergistic effect of and genes.