Multiple origins of spontaneously arising micronuclei in HeLa cells: direct evidence from long-term live cell imaging.
Although micronuclei (MNi) are extensively used to evaluate genotoxic effects and chromosome instability, the most basic issue regarding their origins has not been completely addressed due to limitations of traditional methods. Recently, long-term live cell imaging was developed to monitor the dynamics of single cell in a real-time and high-throughput manner. In the present study, this state-of-the-art technique was employed to examine spontaneous micronucleus (MN) formation in untreated HeLa cells. We demonstrate that spontaneous MNi are derived from incorrectly aligned chromosomes in metaphase (displaced chromosomes, DCs), lagging chromosomes (LCs) and broken chromosome bridges (CBs) in later mitotic stages, but not nuclear buds in S phase. However, most of bipolar mitoses with DCs (91.29%), LCs (73.11%) and broken CBs (88.93%) did not give rise to MNi. Our data also show directly, for the first time, that MNi could originate spontaneously from (1) MNi already presented in the mother cells; (2) nuclear fragments that appeared during mitosis with CB; and (3) chromosomes being extruded into a minicell which fused with one of the daughter cells later. Quantitatively, most of MNi originated from LCs (63.66%), DCs (10.97%) and broken CBs (9.25%). Taken together, these direct evidences show that there are multiple origins for spontaneously arising MNi in HeLa cells and each mechanism contributes to overall MN formation to different extents.
Laboratory of Molecular and Cell Genetics, Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of China.
Xiaotang
X
Rao
Yingyin
Y
Zhang
Qiyi
Q
Yi
Heli
H
Hou
Bo
B
Xu
Liang
L
Chu
Yun
Y
Huang
Wenrui
W
Zhang
Michael
M
Fenech
Qinghua
Q
Shi
D002869:Chromosome Aberrations / Q000145:classification*; D006367:HeLa Cells; D006801:Humans; D007090:Image Interpretation, Computer-Assisted; D008677:Metaphase; D048629:Micronuclei, Chromosome-Defective; D015162:Micronucleus Tests / Q000379:methods; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D013997:Time Factors