Poster H4 - CSRP1 knockdown modulates spheroid cell invasion in human lung cancer cell line H1299

Thursday, October 24, 2024

2:45 PM – 3:45 PM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

Emily Nhan (she/her/hers)

Undergraduate Researcher
University of California Irvine
Fullerton, California, United States

Co-Author(s)

Ashley Urrutia Avila

Graduate Researcher
University of California Irvine, United States

Last Author(s)

Timothy Downing

Associate Professor
University of California, Irvine
Irvine, California, United States

Primary Investigator(s)

MD

Michelle Digman

Associate Professor
University of California Irvine, United States

Introduction: Lung cancer is the leading cause of cancer deaths worldwide due to its great metastatic potential. Studies demonstrate that reduced expression of focal adhesions, which comprise multi-protein structures that connect the cytoskeleton of a cell to their surrounding extracellular matrix (ECM), increases metastatic spread. Additionally, previous studies have found that disrupted cell attachment to the ECM promotes cell survival and invasion potential during metastasis. Preliminary data from our lab demonstrates that CSRP1 gene knockdown (CSRP1 KD), a gene associated in focal adhesion signaling, increases metastatic spread of non-small lung cancer (NSLC) cell line H1299 in immunodeficient NSG mice. Although significant, these preliminary studies require further investigation to better understand the cellular mechanisms driving this increase in metastatic potential upon CSRP1 KD. Thus, the aim of this study is to investigate the fundamental mechanism of metastasis within a simplified in vitro spheroid invasion model assay. We hope to use these findings to better understand the cellular mechanisms driving metastasis. We hypothesize that CSRP1 KD in the H1299 cell line will increase spheroid invasion compared to its control.

Materials and

Methods: To investigate this hypothesis, H1299 cells were treated via lentiviral transduction to knockdown the CSRP1 gene. To create spheroids, transduced H1299 cells were cultured in 96 well plates with ultra-low adhesion and spun down at 1000 rpm for 10 minutes. After 3 days, cells were then transferred into matrigel to mimic the tumor microenvironment. Once transferred cells were imaged over the course of 4 days. These images were then manually outlined using FIJI ImageJ and the area, perimeter length, and circularity of these outlines were quantified.

Results, Conclusions, and Discussions: It was determined that H1299 cells with CSRP1 KD resulted in a higher variance in spheroid area, perimeter length, and circularity. Notably, increased variance in spheroid area upon CSRP1 KD was the result of large increases in invasiveness for a subset of H1299 spheroids. These findings were consistent with our in vivo studies. Future studies will examine the effects of the CSRP1 KD in other NSLC cell lines such as A549 and H460.

Acknowledgements (Optional):