辐射对沉默ATRX的H460细胞增殖以及DNA损伤修复的影响

doi:10.13491/j.issn.1004-714x.2019.04.004

2019, Vol. 28

Issue (4): 364-367 DOI: 10.13491/j.issn.1004-714x.2019.04.004

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The effects of radiation on proliferation and DNA damage repair in H460 cells silenced ATRX

XU Weiqiang¹, TANG Geng¹, LIU Chunyan², LI Xin¹, YANG Yanming³, GONG Shouliang¹, YU Lei³, WANG Zhicheng¹

1. NHC Key laboratory of Radiobiology(Jilin University), School of Public Health, Jilin University, Changchun 130021 China;
2. Department of Radiology, Second Hospital of Jilin University;
3. Department of Radiotherapy, Second Hospital of Jilin University

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Abstract Objective To explore the effects of radiation on proliferation and DNA damage repair, and to clarify the relationship between them.Methods Lentivirus plasmids of shATRX1, shATRX2 and shATRX3 were transfected into 293T cells, H460 cells were infected by lentivirus, shATRX1-H460, shATRX2-H460 and shATRX3-H460 cell lines with low/no ATRX expression were obtained, shControl-H460 cells was used as the control. The silencing efficiency was detected by Western blot. Cell proliferation was measured by colony formation assay, γH2AX and Rad51 foci were detected by immunofluorescence, and the numbers were counted, PARP1, γH2AX and Rad51 expressions were measured by Western blot, respectively.Results ATRX expressed in H460 and shControl-H460 cells, while reduced in shATRX1-H460, shATRX2-H460 and shATRX3-H460 cells. The colony formation assay showed that the SFs of shATRX2-H460 and shATRX3-H460 cells was lower than that of shControl-H460 cells. After shControl-H460 and shATRX3-H460 cells were irradiated by 4 Gy at 1 h, γH2AX foci reached a maximum, while Rad51 foci reached a maximum, at 3 h, and then decreased. Compared with shControl-H460 cells, γH2AX foci at 1 and 6 h, Rad51 foci at 1, 3 and 6 h were significantly increased (P<0.05, P<0.001). And PARP1, γH2AX and Rad51 protein expressions in shATRX3-H460 cells increased at 3 and 6 h compared with that in shContral-H460 cells.Conclusion The cell models of targeted silencing of ATRX in H460 cells were successfully obtained, and the cell proliferation ability was reduced after radiation, which may be related to the reduction of DNA damage repair ability.

Key words： ATRX γH2AX Foci Rad51 Foci DNA Damage Repair Proliferation

Received: 03 February 2019

PACS:

Q691

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	Articles by authors
	XU Weiqiang
	TANG Geng
	LIU Chunyan
	LI Xin
	YANG Yanming
	GONG Shouliang
	YU Lei
	WANG Zhicheng

Cite this article:

XU Weiqiang,TANG Geng,LIU Chunyan, et al. The effects of radiation on proliferation and DNA damage repair in H460 cells silenced ATRX[J]. , 2019, 28(4): 364-367.

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http://www.zgfsws.com/EN/10.13491/j.issn.1004-714x.2019.04.004 OR http://www.zgfsws.com/EN/Y2019/V28/I4/364

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