Article Figures & Data
Figures
- Figure 1.
DFS according to RASSF1A promoter gene methylation status.
- Figure 2.
OS according to RASSF1A and DAPK promoter gene methylation status. A, OS according to RASSF1A. B, OS according to DAPK status. Adj., adjusted.
- Figure 3.
DFSs according to RASSF1A promoter gene methylation status and chemotherapy arm. A, RASSF1A unmethylated patients. B, RASSF1A methylated patients. Adj., adjusted.
- Figure 4.
RASSF1A siRNA-mediated downregulation in HBECs. Immortalized human bronchial epithelial cells are expressing normal amount of RASSF1A protein, and no methylation of RASSF1A promoter was detected in basal conditions (data not shown). A RASSF1A-specific siRNA transfection (right) induced a drastic RASSF1 protein downregulation as shown by immunofluorescence staining with an RASSF1 antibody (right column, right) as compared with the effect of a control irrelevant siRNA (left column, left). Simultaneously, α-tubulin staining of transfected cells showed that RASSF1A downregulation was associated with a drastic tubulin network and cell shape rearrangement from H48 to H96 posttransfection (right column, right) as compared with the lack of effect of the control siRNA (left column, right). Microphotographs shown here are representative of 3 independent experiments.
- Figure 5.
A, Definition of prognosis groups for DFS by stepwise Cox regression. B, definition of prognosis groups for OS by stepwise Cox regression.
Tables
- Table 1.
Characteristics of Bio-IFCT-0002 patients with RASSF1A and DAPK1 promoter gene methylation analysis
All patients RASSF1A methylated RASSF1A unmethylated Pf All patients DAPK1 methylated DAPK1 unmethylated Pf Characteristics (n = 202)a (n = 44) (n = 158) (n = 198)b (n = 99) (n = 99) Sex Male 158 (78.2%) 38 (86.4%) 120 (75.9%) 0.14 154 (77.8%) 80 (80.8%) 74 (74.7%) 0.31 Female 44 (21.8%) 6 (13.6%) 38 (24.1%) 44 (22.2%) 19 (19.2%) 25 (25.3%) Age at inclusion ≤60 y 98 (48.5%) 17 (38.6%) 81 (51.3%) 0.14 95 (48%) 42 (42.4%) 53 (53.5%) 0.12 >60 y 104 (51.5%) 27 (61.4%) 77 (48.7%) 103 (52%) 57 (57.6%) 46 (46.5%) Pack-years ≤10 24 (11.9%) 2 (4.5%) 22 (13.9%) 0.089 24 (12.1%) 12 (12.1%) 12 (12.1%) 1.00 >10 178 (88.1%) 42 (95.5%) 136 (86.1%) 174 (87.9%) 87 (87.9%) 87 (87.9%) WHO PSe 0 166 (82.2%) 38 (86.4%) 128 (81%) 0.41 162 (81.8%) 79 (79.8%) 83 (83.8%) 0.46 1 or 2 36 (17.8%) 6 (13.6%) 30 (19%) 36 (18.2%) 20 (20.2%) 16 (16.2%) Histology SCC 73 (36.1%) 14 (31.8%) 59 (37.3%) 0.50 71 (35.9%) 40 (40.4%) 31 (31.3%) 0.18 Non-SCC 129 (63.9%) 30 (68.2%) 99 (62.7%) 127 (64.1%) 59 (59.6%) 68 (68.7%) Arm (ITT) Gemcitabine 4 cycles PRE 53 (26.2%) 14 (31.8%) 39 (24.7%) 53 (26.8%) 30 (30.3%) 23 (23.2%) Gemcitabine 2 cycles PERI 52 (25.7%) 11 (25%) 41 (25.9%) 51 (25.8%) 26 (26.3%) 25 (25.3%) Paclitaxel 4 cycles PRE 44 (21.8%) 9 (20.5%) 35 (22.2%) 0.80 43 (21.7%) 21 (21.2%) 22 (22.2%) 0.59 Paclitaxel 2 cycles PERI 53 (26.2%) 10 (22.7%) 43 (27.2%) 51 (25.8%) 22 (22.2%) 29 (29.3%) Chemo doublet Gemcitabine 105 (52%) 25 (56.8%) 80 (50.6%) 0.47 104 (52.5%) 56 (56.6%) 48 (48.5%) 0.25 Paclitaxel 97 (48%) 19 (43.2%) 78 (49.4%) 94 (47.5%) 43 (43.4%) 51 (51.5%) No. of cycles 3–4 cycles 87 (43.1%) 18 (40.9%) 69 (43.7%) 0.74 84 (42.4%) 43 (43.4%) 41 (41.4%) 0.77 0-1-2 cycles 115 (56.9%) 26 (59.1%) 89 (56.3%) 114 (57.6%) 56 (56.6%) 58 (58.6%) pStagec 0, I 122 (60.4%) 23 (52.3%) 99 (62.7%) 0.24 121 (61.1%) 60 (60.6%) 61 (61.6%) 0.88 II, III, IV 80 (39.6%) 21 (47.7%) 59 (37.3%) 77 (38.9%) 39 (39.4%) 38 (38.4%) cT 1 51 (25.2%) 9 (20.5%) 42 (26.6%) 0.41 51 (25.8%) 28 (28.3%) 23 (23.2%) 0.42 2 + 3 151 (74.8%) 35 (79.5%) 116 (73.4%) 147 (74.2%) 71 (71.7%) 76 (76.8%) Responsed No 109 (55.1%) 23 (53.5%) 86 (55.5%) 0.82 108 (55.7%) 50 (52.1%) 58 (59.2%) 0.32 Yes 89 (44.9%) 20 (46.5%) 69 (44.5%) 86 (44.3%) 46 (47.9%) 40 (40.8%) -
↵aTwo hundred and two successful RASSF1A methylation results, 6 nondetermined.
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↵bOne hundred and ninety eight successful DAPK methylation results, 10 nondetermined.
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↵cAll c-stages, but 2 noneligible patients with clinical stage IIIA, were I to II (inclusion criteria in the trial).
Pathologic analysis led to reclassification of 7 patients into p-stage IV because of a distant parenchymal metastasis nodule in another lobe (1997 Revised International System for Staging Lung Cancer, used at time of initiation of this trial), but would be classified as p-T4 and p-stage IIIA according to the 2007 7th edition of the staging system.
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↵dFour patients were not evaluable for response.
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↵eWorld Health Organization scores for performance status range from 0 to 2, with a score of 0 indicating no symptoms, 1 mild symptoms, and 2 moderate symptoms.
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↵fP values were calculated by the χ2 test or Fisher exact test when appropriate.
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Supplementary Data
Files in this Data Supplement:
- Supplementary Methods, Figures 1-2, Figure 4, Tables 1-5 - PDF file - 344K, Supplementary methods, suppl. Figure 1 with with the specimens flow-chart, suppl. Figure 2 with the diagram of the clinical trial IFCT 0002 and sampling timepoints, suppl. Figure 4 with RASSF1A interactome, suppl. Table 1 & 2 with primers used in the study, suppl. Table 3 with all the molecular analyses in the bio-IFCT 0002 study, suppl. Table 4 comparing patients with or without molecular analyses, suppl. Table 5 with all the HRs and p-values forthe molecular and clinical variables in the final prognostic analysis
- Supplementary Table 3 - PDF file - 205K, Correlation between RASSF1 promoter gene methylation and RASSF1A protein expression by immunoblotting
- Acknowledgements List - PDF file - 21K, Acknowledegements to IFCT staff (administrative and CRAs), to expert pathological panel, and list of investigators of the phase 3 clinical trial IFCT 0002
Volume 18, Issue 10
