Despite some expression of folate receptor α in human mesothelioma cells, internalization of methotrexate is predominantly carrier mediated Nushmia Z.

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Presentation transcript:

Despite some expression of folate receptor α in human mesothelioma cells, internalization of methotrexate is predominantly carrier mediated Nushmia Z. Khokhar, MDa, Alicia F.Y. Lam, BSa, Valerie W. Rusch, MDb, F.M. Sirotnak, PhDa The Journal of Thoracic and Cardiovascular Surgery Volume 123, Issue 5, Pages 862-868 (May 2002) DOI: 10.1067/mtc.2002.120352 Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Standard curve of CT value against log of starting quantity of β-actin. Dots represent standard samples plotted in triplicate. The slope is −3.37, and the correlation coefficient is 0.998. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 862-868DOI: (10.1067/mtc.2002.120352) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Time course for internalization of 3H methotrexate in various human tumor cells. Cells were incubated with 2 μmol/L 3H methotrexate (MTX) with or without 5 μmol/L folic acid in 50 mmol/L HEPES and 180 mmol/L sucrose brought to pH 7.4 with MgO. Aliquots of cell suspensions were removed at the indicated times for processing. Additional details are provided in the text. Average of 3 experiments with SEM ± 20% or less. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 862-868DOI: (10.1067/mtc.2002.120352) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 Real-time RT-PCR determinations of FRA and RFC1 gene expression in the VAMT-1 human mesothelioma cell line. ΔRn represents the normalized fluorescence signal minus the baseline signal established in the first few cycles of PCR. CT represents the fractional cycle number at which an increase in the reporter fluorescence above a set threshold (horizontal black line) is first detected. β-Actin gene expression was used as a control to normalize these PCR reactions. These data are representative of replicate runs. Additional details are given in the text. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 862-868DOI: (10.1067/mtc.2002.120352) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 Relative levels of RFC1 and FRA mRNA expression in various cell lines. The y axis scale is in relative units (specific target mRNA relative to β-actin mRNA expression). The Journal of Thoracic and Cardiovascular Surgery 2002 123, 862-868DOI: (10.1067/mtc.2002.120352) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 5 Relative levels of RFC1 and FRA expression in mesothelioma tumor samples on the basis of their histologic subtypes. The y axis scale is in relative units (specific target mRNA relative to β-actin mRNA expression). The Journal of Thoracic and Cardiovascular Surgery 2002 123, 862-868DOI: (10.1067/mtc.2002.120352) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions