Effect of lamin-A expression on migration and nuclear stability of ovarian cancer cells

Yixuan Wang; Jing Jiang; Liuqing He; Guanghui Gong; Xiaoying Wu

doi:10.1016/j.ygyno.2018.10.030

Effect of lamin-A expression on migration and nuclear stability of ovarian cancer cells

Gynecol Oncol. 2019 Jan;152(1):166-176. doi: 10.1016/j.ygyno.2018.10.030. Epub 2018 Oct 29.

Authors

Yixuan Wang¹, Jing Jiang², Liuqing He², Guanghui Gong¹, Xiaoying Wu³

Affiliations

¹ Department of Pathology, Xiangya Hospital, Central South University, Changsha, Hunan, China; Department of Pathology, School of Basic Medical Science, Central South University, Changsha, Hunan, China.
² Department of Pathology, School of Basic Medical Science, Central South University, Changsha, Hunan, China.
³ Department of Pathology, Xiangya Hospital, Central South University, Changsha, Hunan, China; Department of Pathology, School of Basic Medical Science, Central South University, Changsha, Hunan, China. Electronic address: xyw2007@csu.edu.cn.

PMID: 30384980
DOI: 10.1016/j.ygyno.2018.10.030

Abstract

Objective: Nuclear lamina plays important roles in nuclear shape and mechanical stability. Many studies demonstrated that defects of lamin-A were associated with several diseases, but little research was found on its potential roles in ovarian cancer.

Methods: GEPIA and GEO database were used to analyze lamin-A in ovarian tissues, followed by assessing lamin-A and prognosis of ovarian cancer patients with Kaplan-Meier plotter. Then, transient transfected HO-8910 cells with shRNA to knockdown lamin-A. Knockdown efficiency was determined by western blot, qRT-PCR and immunofluorescence. Meanwhile, lamin-A was overexpressed in HO-8910 PM cells. Then, 2D migration, 3D migration through 3 μm and 8 μm pores were carried out, followed by immunofluorescence and TEM observation.

Results: Lamin-A tended to be lower in ovarian cancer, and higher expression of lamin-A was associated with better survival. After lamin-A knockdown, 2D and 3D migration (3 μm, 8 μm) abilities of HO-8910 cells were significantly increased (p < 0.001), while overexpression of lamin-A in HO-8910PM impeded migration. Meanwhile, when HO-8910 cells migrated through 3 μm pores, nuclei became strikingly elongated, and down-regulation of lamin-A promoted nuclear plasticity, making the circularity of nucleus increased. Besides, further knockdown group had the highest proportion of γ-H2AX, with micronuclei forming. Furthermore, western blot showed that the expression of BRCA1, Ku80 and Rad50 decreased significantly after further knockdown, suggesting impairment of DNA damage repair.

Conclusions: Lamin-A was down-regulated in ovarian cancer, and higher lamin-A was associated with better prognosis. Nuclei with high lamin-A were severely deformed through constricted pores. Moderate lamin-A enhanced nuclear plasticity, so as to strengthen migration ability. When lamin-A was further knockdown, ovarian cancer cells that migrated through restricted pores decreased, with DNA damage, genomic instability and impairment of DNA damage repair.

Keywords: DNA damage; Lamin-A; Migration; Nucleus; Ovarian cancer.

MeSH terms

Cell Line, Tumor
Cell Movement
Cell Nucleus / pathology*
Cell Nucleus / ultrastructure
DNA Damage
Female
Histones / analysis
Humans
Lamin Type A / analysis
Lamin Type A / physiology*
Ovarian Neoplasms / genetics
Ovarian Neoplasms / pathology*

Substances

H2AX protein, human
Histones
Lamin Type A