Simulation of P53-dependent cell death in murine intestinal stem cells <italic style="font-style: italic">in vitro</italic> by X-ray radiation

Xiaoqin Li; Yinhua Fang; Jiong Ren; Caizhi Tang; Xinze Ran; Yongping Su; Fengchao Wang; Hongying Cheng

doi:10.16016/j.1000-5404.201612080

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Simulation of P53-dependent cell death in murine intestinal stem cells in vitro by X-ray radiation

更新时间：2026-01-09

- Simulation of P53-dependent cell death in murine intestinal stem cells in vitro by X-ray radiation
- Journal of Third Military Medical University Vol. 39, Issue 9, Pages: 865-871(2017)
- 作者机构：
- 作者简介：
- 基金信息：
  
  the General Program of National Science Foundation of China(81472916);the Major Program of the "Twelfth Five-year Plan" for Medical Development of PLA(AWS13J002);the Independent Research Project of State Key Laboratory(SKLZZ201420)
- DOI：10.16016/j.1000-5404.201612080
  CLC：
- Received：12 December 2016，
  
  Published：15 May 2017
- 稿件说明：
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Xiaoqin Li, Yinhua Fang, Jiong Ren, et al. Simulation of P53-dependent cell death in murine intestinal stem cells in vitro by X-ray radiation[J]. Journal of Third Military Medical University, 2017, 39(9): 865-871.
DOI：

Xiaoqin Li, Yinhua Fang, Jiong Ren, et al. Simulation of P53-dependent cell death in murine intestinal stem cells in vitro by X-ray radiation[J]. Journal of Third Military Medical University, 2017, 39(9): 865-871. DOI： 10.16016/j.1000-5404.201612080.

摘要

Abstract

Objective

To estab

lish an

in vitro

irradiation model in murine intestinal stem cells (ISCs) with 6 Gy X-ray radiation in order to mimic the repair process of radiation-induced

in vivo

P53-dependent ISC death.

Methods

The small intestinal crypts were isolated from wide type (WT) and P53

-/-

mice

and then cultured in a matrigel 3D system for 24 h. Then the crypt clones were irradiated by X-ray at a dose of 6 Gy. The growth and death of crypt clones were observed in 6 d after irradiation. The expression levels of P53 targeting genes

Bax

Bbc3

Mdm2

P21 and Bcl2l1

and ISCs specific genes Lgr5

Olfm4 and Ascl2 in the WT crypt clones at different time points within 24 h after irradiation were measured by quantitative real-time PCR (RT-PCR).

Results

The crypts clones derived from WT and P53

-/-

mice could bud in 48 h after embedding in matrigel and form the organoids in 96 to 120 h. Irradiation of 6 Gy X-ray resulted in massive death of crypt clones from WT mice within 24 h

and total blocking in the budding of crypt clone

with only (43.84±5.73)% clones survival. But it had no effect on the budding of the crypt clones from P53

-/-

mice

and led to (83.69±5.89)% clones survival

significantly higher than that in the WT crypt colons (

0.01). In the WT crypt clone

the P53 downstream genes were activated immediately after irradiation

and maintained in high levels within 24 h

while the levels of ISCs specific gens were decreased rapidly within 12 h

and reached the lowest in 24 h after irradiation exposure.

Conclusion

Intestinal crypt 3D culture model exposure to 6 Gy irradiation can well mimic P53-dependent massive crypt clone death and regeneration

in vivo

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Keywords

references

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⁰

Simulation of P53-dependent cell death in murine intestinal stem cells in vitro by X-ray radiation

DOI：10.16016/j.1000-5404.201612080

摘要

Abstract

关键词

Keywords

references