Role and mechanism of RNA demethylase FTO in apoptosis of GC-2 cells induced by bisphenol S

Jingzhi LI; Wenbin LIU; Shimeng ZHOU; Hongqiang CHEN; Jia CAO

doi:10.16016/j.2097-0927.202111016

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Role and mechanism of RNA demethylase FTO in apoptosis of GC-2 cells induced by bisphenol S

Public Health and Preventive Medicine | 更新时间：2026-01-09

- Role and mechanism of RNA demethylase FTO in apoptosis of GC-2 cells induced by bisphenol S
- Journal of Army Medical University Vol. 44, Issue 6, Pages: 601-610(2022)
- 作者机构：
  
  1.453001 河南新乡，新乡医学院公共卫生学院
  2.400038 重庆，陆军军医大学(第三军医大学)军事预防医学系：毒理学研究所
  3.400038 重庆，陆军军医大学(第三军医大学)军事预防医学系：军队环境卫生学教研室
  4.110122 沈阳，中国医科大学公共卫生学院
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(82130097);the National Key Research and Development Program of China(2017YFC1002001)
- DOI：10.16016/j.2097-0927.202111016
  CLC： R321.1;R329.25;R994.6
- Received：02 November 2021，
  
  Revised：2022-01-07，
  
  Published：30 March 2022
- 稿件说明：
移动端阅览
Jingzhi LI, Wenbin LIU, Shimeng ZHOU, et al. Role and mechanism of RNA demethylase FTO in apoptosis of GC-2 cells induced by bisphenol S[J]. Journal of Army Medical University, 2022, 44(6): 601-610.
DOI：

Jingzhi LI, Wenbin LIU, Shimeng ZHOU, et al. Role and mechanism of RNA demethylase FTO in apoptosis of GC-2 cells induced by bisphenol S[J]. Journal of Army Medical University, 2022, 44(6): 601-610. DOI： 10.16016/j.2097-0927.202111016.

摘要

目的

探讨RNA去甲基化酶FTO在双酚S(bisphenol S，BPS)诱导小鼠精母细胞GC-2凋亡中的作用及机制。

方法

采用终浓度为0(DMSO对照组)、20、40、80、160、320 μmol/L的BPS处理小鼠精母细胞株GC-2细胞72 h，镜下观察细胞形态变化；采用CCK-8法检测细胞活力，通过流式细胞术检测细胞凋亡和周期改变；采用比色法检测RNA甲基化的总体水平；通过Western blot检测RNA甲基转移酶METTL3、RNA去甲基化酶FTO、RNA甲基化识别蛋白YTHDF1和细胞凋亡相关分子P53、P21、BCL2、Caspase-9、Caspase-3、BAX以及细胞周期相关分子CyclinD1、CDK4、CDK2的蛋白表达水平。建立FTO基因干扰细胞模型，采用CCK-8法检测细胞活力，采用流式细胞术检测细胞凋亡改变，通过Western blot检测FTO和细胞凋亡相关分子P53、BCL2、BAX的蛋白表达水平。

结果

与对照组相比，BPS染毒组GC-2细胞数量明显减少，出现较多的空泡细胞；细胞存活率呈下降趋势，尤其在160、320 μmol/L BPS染毒剂量下降显著(

0.05)；细胞凋亡率显著升高，凋亡相关分子P53、P21、Caspase-9和Caspase-3蛋白水平显著增加，BCL2的蛋白水平显著下调(

0.05)；BPS浓度在80、160、320 μmol/L时细胞周期阻滞在G

/S期，细胞周期G

/S期相关分子CyclinD1、CDK4、CDK2的蛋白水平显著下调(

0.05)；在80~320 μmol/L BPS染毒剂量组下，RNA甲基化的总体水平显著升高(

0.01)；RNA甲基转移酶METTL3和RNA甲基化识别蛋白YTHDF1的蛋白水平无明显变化，而RNA去甲基化酶FTO的蛋白水平显著下降(

0.05)。与对照组相比，干扰FTO表达后GC-2细胞数量明显减少，出现较多的空泡细胞，细胞活力显著降低(

0.01)；细胞凋亡率显著升高，P53的蛋白水平显著上调，BCL2的蛋白水平显著下调(

0.05)。

结论

RNA去甲基化酶FTO可能通过调控P53-P21和BCL2线粒体途径在BPS诱导的GC-2细胞凋亡过程中发挥重要作用。

Abstract

Objective

To explore the role and mechanism of RNA demethylase FTO in the apoptosis of mouse spermatocyte GC-2 cells induced by bisphenol S (BPS).

Methods

GC-2 cells were treated with BPS at a final concentration of 0 (DMSO control group)

160

and 320 μmol/L for 72 h. The cell morphology changes were observed with microscopy. CCK-8 assay was utilized to detect cell viability

and flow cytometry was used to detect cell apoptosis and cycle changes. Colorimetric assay was adopted to measure the global level of m6A RNA methylation. Western blotting was employed to detect the protein levels of RNA methyltransferase METTL3

RNA demethylase FTO

RNA binding protein

YTHDF1

apoptosis-related molecules P53

P21

BCL2

Caspase9

Caspase3

BAX and cell cycle-related molecules CyclinD1

CDK4

CDK2. After FTO gene interference cell model was established

CCK-8 assay was used to measure cell survival and flow cytometry to detect cell apoptosis. The protein levels of FTO

P53

BCL2 and BAX were detected by Western blotting.

Results

Compared with the control group

the number of GC-2 cells was significantly reduced and more vacuole cells appeared in the BPS exposure group. The cell survival rate was significantly decreased and the apoptotic rate was markedly increased at 160 and 320 μmol/L of BPS (

0.05). The protein levels of apoptosis-related molecules P53

P21

Caspase9 and Caspase3 were increased significantly

and that of BCL2 was decreased significantly (

0.05). When the BPS concentration was 80

160 and 320 μmol/L

cell cycle was arrested at G

/S phase (

0.05). The protein levels of G

/S phase related molecules CyclinD1

CDK4

and CDK2 were significantly down-regulated (

0.05). When the dose of BPS increased to 80~320 μmol/L

the global RNA methylation level was relatively increased (

0.01). The protein levels of RNA methyltransferase METTL3 and RNA binding protein YTHDF1 did not change obviously

while that of RNA demethylase FTO decreased sharply (

0.05). Compared with the control group

after interference with FTO expression

the number and viability of GC-2 cells were significantly reduced

and the apoptotic rate was notably increased (

0.05). The protein level of P53 was remarkably up-regulated

and that of BCL2 down-regulated when FTO expression was knocked down (

0.05).

Conclusion

RNA demethylase FTO may play an important role in BPS-induced apoptosis of GC-2 cells through regulating P53-P21 and BCL2 mitochondrial pathway.

关键词

Keywords

references

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