T24 HRAS transformed NIH/3T3 mouse cells (GhrasT-NIH/3T3) in serial tumorigenic in vitro/in vivo passages give rise to increasingly aggressive tumorigenic cell lines T1-A and T2-A and metastatic cell lines T3-HA and T4-PA

Journal Article · Fri Jan 01 00:00:00 EST 2016 · Experimental Cell Research

DOI:https://doi.org/10.1016/J.YEXCR.2015.07.029· OSTI ID:22746393

Lam, Tan ^[1]; McElhinney, Aaron ^[2]; Anders, Joel ^[2]; Barcus, Scott ^[2]; others, and

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1. Department of Biochemistry, Oral Roberts University School of Medicine, 7777 S Lewis Ave, Tulsa, OK 74171 (United States)
2. Department of Biology, Grove City College, 100 Campus Drive, Grove City, PA 16127 (United States)

Cancer cells often arise progressively from “normal” to “pre-cancer” to “transformed” to “local metastasis” to “metastatic disease” to “aggressive metastatic disease”. Recent whole genome sequencing (WGS) and spectral karyotyping (SKY) of cancer cells and tumorigenic models have shown this progression involves three major types of genome rearrangements: ordered small step-wise changes, more dramatic “punctuated evolution” (chromoplexy), and large catastrophic steps (chromothripsis) which all occur in random combinations to generate near infinite numbers of stochastically rearranged metastatic cancer cell genomes. This paper describes a series of mouse cell lines developed sequentially to mimic this type of progression. This starts with the new GhrasT-NIH/Swiss cell line that was produced from the NIH/3T3 cell line that had been transformed by transfection with HRAS oncogene DNA from the T24 human bladder carcinoma. These GhrasT-NIH/Swiss cells were injected s.c. into NIH/Swiss mice to produce primary tumors from which one was used to establish the T1-A cell line. T1-A cells injected i.v. into the tail vein of a NIH/Swiss mouse produced a local metastatic tumor near the base of the tail from which the T2-A cell line was established. T2-A cells injected i.v. into the tail vein of a nude NIH/Swiss mouse produced metastases in the liver and one lung from which the T3-HA (H=hepatic) and T3-PA (P=pulmonary) cell lines were developed, respectively. T3-HA cells injected i.v. into a nude mouse produced a metastasis in the lung from which the T4-PA cell line was established. PCR analysis indicated the human T24 HRAS oncogene was carried along with each in vitro/in vivo transfer step and found in the T2-A and T4-PA cell lines. Light photomicrographs indicate that all transformed cells are morphologically similar. GhrasT-NIH/Swiss cells injected s.c. produced tumors in 4% of NIH/Swiss mice in 6–10 weeks; T1-A cells injected s.c. produced tumors in 100% of NIH/Swiss mice in 7–10 days. T1-A, T-2A, T3-HA and T4-PA cells when injected i.v. into the tail produced local metastasis in non-nude or nude NIH/Swiss mice. T4-PA cells were more widely metastatic than T3-HA cells when injected i.v. into nude mice. Evaluation of the injected mice indicated a general increase in metastatic potential of each cell line in the progression as compared to the GhrasT-NIH/3T3 transformed cells. A new photomicrographic technique to follow growth rates within six preselected 2×2 mm{sup 2} grids per plate is described. Average doubling times of the transformed cells GhrasT-NIH/3T3 (17 h), T1A (17.5 h), T2A (15.5 h), T3-HA (17.5 h) and T4-PA (18.5 h) (average 17.2 h) were significantly faster (P=0.006) than NIH Swiss primary embryonic cells and NIH/3T3 cells (22 h each). This cell series is currently used in this lab for studies of cancer cell inhibitors, mitochondrial biogenesis and gene expression and is available for further study by other investigators for intra- and inter-laboratory comparisons of WGS, transcriptome sequencing, SKY and other analyses. The genome rearrangements in these cells together with their phenotypic properties may help provide more insights into how one tumorigenic progression occurred to produce the various cell lines that led to the highly metastatic T4-PA cell line. - Highlights: • A tumor progression model derived from a new T24 HRAS transformed NIH/3T3 cell line. • T1-A, T2-A, T3-HA, T4PA lines derived from GhrasT-NIH/3T3, T1-A, T2-A, T3-HA tumors. • PCR detected the human HRAS oncogene in T2-A and T4-PA metastatic tumors. • These five new cell lines showed a progressive increase in tumorigenic potential. • New method obtained doubling times using daily photomicrographs of gridded plates.

• OSTI ID: 22746393
• Journal Information: Experimental Cell Research, Vol. 340, Issue 1; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0014-4827
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
BLADDER
CARCINOMAS
COMPARATIVE EVALUATIONS
DNA
EMBRYONIC CELLS
HYDROGEN 5
IN VITRO
IN VIVO
INDIUM 100
INTERLABORATORY COMPARISONS
KARYOTYPE
LIVER
LUNGS
METASTASES
MICE
MITOCHONDRIA
ONCOGENES
PLANT GROWTH
POLYMERASE CHAIN REACTION
RANDOMNESS
RISE
STOCHASTIC PROCESSES
VEINS
VISIBLE RADIATION