Article Figures & Data
Figures
- Fig. 1.
Sensitivity to a COX-2 inhibitor, nimesulide, measured by MTT assay (A) and in relation to COX-2 expression (B). A, dose-dependent inhibition of the proliferation of NSCLC lines in response to increasing amounts of nimesulide. ACC-LC-33, -319, -323, and -91 as well as PC-10 are NSCLC cell lines, ACC-LC-172 and -48 are SCLC cell lines, and BEAS2B and HPL1D are normal lung epithelial cell lines. Each data point represents the mean of quadruplicate determinations (bars, SE) in a representative experiment. Similar results were obtained in four independent experiments. B, Northern blot analysis of COX-2 expression using 10μ g of total RNA. A substantial association between sensitivity to nimesulide and COX-2 expression levels is observed. 28S rRNAs, which are visualized by ethidium bromide staining, show loading of similar amounts of RNA in each lane.
- Fig. 2.
Detection of apoptosis induced by incubation with nimesulide. A, detection of apoptosis with the TUNEL method. TUNEL-positive ACC-LC-91 cells are detectable even 6 h after the initiation of nimesulide treatment at 100 μm, whereas a significant increase in the proportion of TUNEL-positive cells is evident at 24 h. Similarly, induction of apoptosis can be seen in nimesulide-treated ACC-LC-323 cells, whereas virtually no effects are observed in BEAS2B cells. B, detection of DNA fragmentation. DNA fragmentation, a hallmark of apoptosis, is evident in ACC-LC-91 cells but not in BEAS2B cells, both of which were treated with nimesulide at concentrations of 10 μm and 100 μm for 48 h. Similar results were obtained in three independent experiments.
- Fig. 3.
Western blot analysis of bcl-2 and bax in relation to nimesulide treatment. Expression levels of bcl-2 and bax in ACC-LC-91 are not changed by the treatment with nimesulide at 100μ m for up to 8 h.
- Fig. 4.
IC50 isobolograms of the use of nimesulide as an adjunct with SM-5887–13-OH, SN-38, taxotere, VP-16, or CDDP in ACC-LC-91. Supra-additive enhancement of chemosensitivity to SM-5887 and SN-38 is seen as a result of the adjunct use of nimesulide at both 10 μm and 30 μm, whereas the combination of taxotere and 30 μm nimesulide shows a nearly synergistic effect. Solid and broken lines represent Mode I and Mode II in the IC50 isobologram, which were constructed from dose-response curves of nimesulide and each chemotherapeutic agent, respectively (24 , 25) . The area delineated by Mode I and Mode II lines represents the envelope of additivity, whereas the area below this envelope represents the supra-additive area and the area above it the sub-additive area.
Tables
- Table 1
Chemosensitivity to five anticancer agents used in combination with nimesulide
Cell line SM-5887-13-OH SN-38 Taxotere VP-16 CDDP IC50 (μg/ml) RRa IC50 (μg/ml) RR IC50 (μg/ml) RR IC50 (μg/ml) RR IC50 (μg/ml) RR NSCLC cell lines ACC-LC-91 0.015 ± 0.003b 1.00 0.0065 ± 0.0010 1.00 0.0032 ± 0.0004 1.00 0.21 ± 0.03 1.00 2.51 ± 0.42 1.00 ACC-LC-91/NIM 10c 0.008 ± 0.002 0.53 0.0039 ± 0.0008 0.60 0.0025 ± 0.0003 0.78 0.17 ± 0.02 0.80 2.11 ± 0.26 0.84 ACC-LC-91/NIM 30c 0.006 ± 0.002 0.40 0.0015 ± 0.0003 0.23 0.0015 ± 0.0002 0.46 0.14 ± 0.02 0.66 1.90 ± 0.23 0.75 ACC-LC-323 0.025 ± 0.004 1.00 0.0055 ± 0.0008 1.00 0.0028 ± 0.0002 1.00 0.24 ± 0.02 1.00 0.45 ± 0.06 1.00 ACC-LC-323/NIM 10 0.023 ± 0.003 0.92 0.0050 ± 0.0007 0.90 0.0022 ± 0.0003 0.78 0.21 ± 0.03 0.87 0.37 ± 0.04 0.82 ACC-LC-323/NIM 30 0.021 ± 0.003 0.84 0.0043 ± 0.0006 0.78 0.0017 ± 0.0003 0.60 0.20 ± 0.02 0.83 0.35 ± 0.03 0.77 ACC-LC-319 0.038 ± 0.005 1.00 0.0061 ± 0.0009 1.00 0.0060 ± 0.0009 1.00 0.50 ± 0.09 1.00 2.10 ± 0.33 1.00 ACC-LC-319/NIM 10 0.034 ± 0.004 0.89 0.0055 ± 0.0008 0.90 0.0051 ± 0.0008 0.85 0.45 ± 0.08 0.90 2.01 ± 0.28 0.95 ACC-LC-319/NIM 30 0.028 ± 0.004 0.73 0.0049 ± 0.0009 0.80 0.0042 ± 0.0009 0.70 0.43 ± 0.09 0.86 1.89 ± 0.24 0.90 ACC-LC-33 0.080 ± 0.010 1.00 0.0091 ± 0.0010 1.00 0.0082 ± 0.0009 1.00 0.82 ± 0.11 1.00 2.88 ± 0.38 1.00 ACC-LC-33/NIM 10 0.078 ± 0.009 0.97 0.0088 ± 0.0009 0.96 0.0073 ± 0.0008 0.89 0.80 ± 0.10 0.97 2.68 ± 0.35 0.93 ACC-LC-33/NIM 30 0.064 ± 0.009 0.80 0.0078 ± 0.0009 0.85 0.0070 ± 0.0009 0.85 0.73 ± 0.09 0.89 2.43 ± 0.40 0.84 Normal lung epithelial cell lines BEAS2B 0.024 ± 0.003 1.00 0.0061 ± 0.0007 1.00 0.0062 ± 0.0006 1.00 0.31 ± 0.02 1.00 1.31 ± 0.22 1.00 BEAS2B/NIM 10 0.023 ± 0.004 0.95 0.0064 ± 0.0005 1.04 0.0070 ± 0.0008 1.12 0.32 ± 0.03 1.03 1.52 ± 0.34 1.16 BEAS2B/NIM 30 0.025 ± 0.002 1.04 0.0064 ± 0.0006 1.04 0.0064 ± 0.0008 1.03 0.33 ± 0.02 1.06 1.50 ± 0.26 1.14 HPL1D 0.042 ± 0.005 1.00 0.0110 ± 0.0010 1.00 0.0092 ± 0.0009 1.00 0.64 ± 0.08 1.00 1.73 ± 0.12 1.00 HPL1D/NIM 10 0.040 ± 0.006 0.95 0.0099 ± 0.0008 0.90 0.0088 ± 0.0011 0.95 0.60 ± 0.07 0.93 1.71 ± 0.21 0.98 HPL1D/NIM 30 0.040 ± 0.007 0.95 0.0098 ± 0.0007 0.89 0.0090 ± 0.0010 0.97 0.59 ± 0.10 0.92 1.92 ± 0.10 1.10 -
a RR, relative resistance.
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b Mean ± SD of the results obtained in three independent experiments.
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c NIM 10 and NIM 30, simultaneous exposure to 10 μm and 30 μm nimesulide, respectively. As a single agent, nimesulide yielded 15% growth inhibition at 10μ m and 28% at 30 μm in ACC-LC-91 cells, 12% and 23% in ACC-LC-323 cells, 8% and 13% in ACC-LC-319 cells, 6% and 10% in ACC-LC-33 cells, 0% and 0% in BEAS2B cells, and 0% and 1% in HPL1D cells (see Fig. 1<$REFLINK> ).
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- Table 2
Radiation sensitivity in combination with nimesulide
Cell line SF1a SF2a SF0a NSCLC cell lines ACC-LC-91 0.90 ± 0.01 0.81 ± 0.02 1.00 ACC-LC-91/NIM 10 0.79 ± 0.03 0.72 ± 0.03 0.85 ± 0.02 ACC-LC-91/NIM 30 0.72 ± 0.03 0.69 ± 0.03 0.80 ± 0.02 ACC-LC-323 0.91 ± 0.02 0.80 ± 0.02 1.00 ACC-LC-323/NIM 10 0.85 ± 0.02 0.74 ± 0.03 0.91 ± 0.01 ACC-LC-323/NIM 30 0.68 ± 0.03 0.63 ± 0.03 0.84 ± 0.02 ACC-LC-319 0.98 ± 0.01 0.97 ± 0.01 1.00 ACC-LC-319/NIM 10 0.90 ± 0.02 0.90 ± 0.02 0.92 ± 0.02 ACC-LC-319/NIM 30 0.88 ± 0.02 0.87 ± 0.02 0.90 ± 0.02 ACC-LC-33 0.87 ± 0.02 0.83 ± 0.02 1.00 ACC-LC-33/NIM 10 0.80 ± 0.03 0.77 ± 0.03 0.95 ± 0.01 ACC-LC-33/NIM 30 0.72 ± 0.03 0.67 ± 0.03 0.91 ± 0.02 Normal lung epithelial cell lines BEAS2B 0.90 ± 0.01 0.82 ± 0.02 1.00 BEAS2B/NIM 10 0.91 ± 0.02 0.84 ± 0.02 1.01 ± 0.01 BEAS2B/NIM 30 0.90 ± 0.02 0.85 ± 0.02 0.99 ± 0.01 HPL1D 0.89 ± 0.01 0.84 ± 0.02 1.00 HPL1D/NIM 10 0.90 ± 0.02 0.85 ± 0.02 0.99 ± 0.01 HPL1D/NIM 30 0.90 ± 0.02 0.84 ± 0.02 0.98 ± 0.02 -
a Surviving fractions for 1 Gy, 2 Gy, and 0 Gy.
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b The values represent means ± SD of a representative experiment. Similar results were obtained in three independent experiments.
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Volume 6, Issue 5
