Our Unique Cell Lines Derived Serially After Injection With Human T24 HRAS Transformed NIH/3T3 Mouse Cells (GhrasT‐NIH/3T3) Showed The Highly Tumorigenic T1‐A Cells Cultured From The First Primary Tumor Had A Six‐Fold Higher LD50 For Resveratrol Killing Than The LD50 For Highly Metastatic Lung T4‐PA Cells Derived After Three Subsequent In Vivo / In Vitro Serial Manipulations

Abstract

Our new cell lines demonstrating progressively increasingly metastatic behavior as they were derived in a series of in vivo/in vitro manipulations (Figure 1) provide the opportunity to evaluate in vivo and in vitro compounds and methods to kill primary tumor cells and various forms of increasingly metastatic cells derived from the same primary tumor (Figure 2) (Ray, D.B., et. al., Experimental Cell Research, 2016 Jan; 340 (1): pp. 1–11). We determined the dose of the anti‐cancer natural phytoalexin resveratrol to kill 50% (LD50s) of our various cells that represent progressive cancer stages. Cell counts were recorded daily, over the course of 3 to 6 days, using gridded plates and photomicrographs of six pre‐determined 2 mm square sections of each plate. An average percent change in cell number from each individual grid was then calculated. P‐values were determined between each treatment time and dose and significance was determined. The NIH Swiss and NIH/3T3 cells were not significantly affected by 3–300 μM after 3 days. After 3 or 4 days the LD50s for the tumorigenic cells were GhrasT‐NIH/3T3 cells: LD50 = 3.5+/− 1.0 μM; T1‐A cells: LD50 = 45 +/− 5.0 μM; T4‐PA cells: LD50 = 7.0 +/− 1.5 μM. Thus, unexpectantly, the highly tumorigenic T1‐A cells were significantly more difficult to kill than the highly metastatic T4‐PA cells. This pattern may mimic the epithelial to mesenchymal transition (EMT) seen in breast and other cancer progression. EMT enhances mobility, invasion, and resistance to apoptotic stimuli as tumor cells acquire stem cell properties and exhibit marked therapeutic resistance. RNA‐Seq, genome sequencing (WGS) and spectral karyotyping (SKY) analysis of these cells demonstrating increasingly aggressive metastatic cancer tumorigenesis that progressed from a single nuclear genotype and mitochondrial haplotype should help contribute to the understanding of cancer progression. These cells are available by contacting our laboratory.Support or Funding InformationSupported in part by the National Science Foundation grant # 8602787, the Grove City College Research Fund, the Grove City College Swezey Research Fund and the Grove City College Janicki Foundation Research Fund.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.