Research Paper|Volume 12, Issue 20|pp 20493—20511

RNA-seq reveals the diverse effects of substrate stiffness on epidermal ovarian cancer cells

Xiaoxu Yang¹, Guohui Wang¹, Xiaolei Huang¹, Min Cheng², Yangyang Han¹

¹School of Life Science and Technology, Weifang Medical University, Weifang 261053, Shandong, P.R. China
²Department of Physiology, Weifang Medical University, Weifang 261053, Shandong, P.R. China

* Equal contribution

Received: May 4, 2020Accepted: July 21, 2020Published: October 22, 2020

Copyright: © 2020 Yang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Background: Increasing evidence has confirmed that ovarian cancer is a mechanically responsive tumor both in vivo and in vitro. However, an understanding of the complete molecular mechanism involved in the response to substrate stiffness is lacking, as the associated transcriptome-wide effects have not been mapped. This limited understanding has restricted the identification of potential mechanically responsive targets in ovarian cancer.

Results: To address these limitations, we used a polyacrylamide hydrogel system with a tunable Young’s modulus that broadly ranged from soft (1 kPa) to normal (6 kPa) and stiff (60 kPa) and investigated the effect of substrate rigidity on the morphology, spreading area, and cytoskeleton of SKOV-3 epidermal ovarian cancer (EOC) cells. RNA-seq analysis of these cells was then performed at appropriate timepoints to map the transcriptome-wide changes associated with stiffness sensing. We identified a large number of stiffness-sensing genes as well as many genes that were enriched in cancer-related pathways. Informed by these diverse expression results and based on bioinformatics analysis, we evaluated the hypothesis that PLEC and TNS2, which are located in focal adhesions and regulated by lnc-ZNF136, may play key roles in the EOC response to substrate stiffness.

Conclusion: Overall, the results of the present study reveal previously unknown features of the EOC stiffness response and provide new insights into EOC metastasis in the clinic.