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Examining the relationship between MCPH1 with H2AX in esophageal cancer cells ECA109

Hongyun Shi, Ke Zhang, Ming Wen, Yu Yuan

Objective: To discover the role of MCPH1 in the DNA double-strand damage induced by ionizing radiation and the relationship with H2AX in esophageal cancer cell ECA109.

Methods: ECA109 cancer cells will be accepted 8 Gy 1 h after irradiation for protein extraction and immunofluorescence observed MCPH1 and H2AX protein expression and nuclear foci changes. Establish a stable low expression of H2AX cell lines. Detecting MCPH1 and H2AX protein expression and nuclear foci changes induced by ionizing radiation after silence H2AX.

Results: 1. Successfully constructed the ECA109 cell line with silence H2AX gene. 2. Ionizing radiation could cause r-H2AX and MCPH1 protein expression increase, as the same as nuclear focus increase of r- H2AX and MCPH1. 3. The protein level and nucleus focus of r-H2AX and MCPH1 are significantly reduced in ECA109 after silence H2AX.

Conclusion: MCPH1 is the part of DNA damage response triggered by ionizing radiation and is located in the damage response downstream and can be regulated by H2AX.