Chapter 25 The Stathmokinetic Experiment: A Single-Parameter and Multiparameter Flow Cytometric Analysis

Traganos, Frank; Kimmel, Marek

doi:10.1016/s0091-679x(08)60530-0

Cited by 4 publications

(2 citation statements)

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“…Stathmokinetic analysis of 3T6R.3-19 cells (relative dhfr copy number 13X) and the parental 3T6 cell line (Figure 4) demonstrates that the cells with amplified dhfr genes are very slow to exit G 1 compared to the parental cells. The slopes of the lines for G 1 exit indicate a mean G 1 duration of about 2 h for the parental 3T6 cells and a mean G 1 duration of about 12 h for the 3T6R.3-19 line (Traganos & Kimmel 1990). The 3T6R.3-19 cells are also slow to transit S-phase and slow to accumulate in G 2 /M.…”

Section: T6 Seriesmentioning

confidence: 93%

“…Stathmokinetic analysis was done using acridine orange multiparameter flow cytometry (Darzynkiewicz, Traganos & Kimmel 1986, Traganos & Kimmel 1990. The use of the metaphase arresting agent, vinblastine, prevents mitosis so that the cells accumulate at this stage of the cell cycle, and cannot re-enter the cell cycle as G 1 cells.…”

Section: Stathmokinetic Analysismentioning

confidence: 99%

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Unbalanced growth in mouse cells with amplified dhfr genes

et al. 1997

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When grown in the absence of methotrexate, cells carrying unstably amplified dihydrofolate reductase (dhfr) genes have a growth disadvantage that is a function of their level of gene amplification. Although this growth disadvantage is thought to drive the loss of unstably amplified dhfr genes in the absence of methotrexate, its mechanism is not understood. The present studies of murine cell lines with different levels of dhfr gene amplification demonstrate that such cells experience increased unbalanced growth (excess RNA and protein content relative to DNA content) with increased levels of dhfr gene amplification. Stathmokinetic analysis of a cell line with unstably amplified dhfr genes showed that the unbalanced growth was associated with a very low rate of G1/S transit, which suggests that amplified DNA sequences may activate a cell cycle checkpoint at the G1/S boundary. Hydroxyurea, which is known to induce rapid elimination of amplified genes at sub-cytotoxic concentrations, also inhibits the cell cycle at the G1/S transition and causes unbalanced growth. Earlier work has shown that hydroxyurea selectively targets those cells within the heterogeneous drug resistant cell populations which have the highest amplified gene dosage. The finding that unstable gene amplification and hydroxyurea have similar effects on the cell suggests that hydroxyurea may achieve this selective targeting by pushing those cells with the highest levels of gene amplification over a critical stress threshold to cause growth arrest or cell death.

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Section: T6 Seriesmentioning

confidence: 93%

Section: Stathmokinetic Analysismentioning

confidence: 99%