Article Figures & Data
Figures
- Fig. 1.
A, immunoblot analysis for characterization of the affinity-purified anti-4N1K peptide antibody. Ten μg of purified TSP-1 were treated with either 2.5 microunits of the purified catalytic domain of MMP3 or left untreated at 37°C for 60 min and electrophoresed, followed by electrotransfer onto PVDF membranes. TSP1 and 4N1K-containing proteins (arrowheads) were visualized by anti-4N1K antibody (left panel). After stripping of membranes, the anti-4N1K antibody, preincubated with 4N1K peptide-immobilized agarose beads for 30 min at 4°C, was used as the first antibody (right panel). B, positive staining of 4N1K-containing proteins in tumor tissue disappeared when the antibody was preincubated with 4N1K peptide-immobilized agarose beads. Sequential tissue sections of RCC were deparaffinized, treated with trypsin, and stained with either anti-4N1K antibody or the antibody preincubated with 4N1K peptide-immobilized beads. Reproducible results were obtained from two independent experiments. ×200.
- Fig. 2.
Detection of TSP1 protein or 4N1K-containing proteins in formalin-fixed, paraffin-embedded tissues by immunohistochemical technique. A, positive staining for 4N1K immunoreactivity in tumor tissue. Deparaffinized specimens were treated with 1 ng/ml trypsin at 37°C for 3 min and incubated with rabbit affinity-purified anti-4N1K antibody. B, the same tissue sections shown in A were incubated with preimmunized rabbit IgG as a negative control. C, positive staining for TSP1 immunoreactivity in tumor tissue. Deparaffinized specimens were heated at 95°C for 40 min and incubated with mouse monoclonal anti-TSP1 antibody. D, the same tissue sections shown in C were incubated with normal mouse IgG as a negative control.
- Fig. 3.
Relationship between the largest diameter of each tumor and immunohistochemical staining for TSP1 or 4N1K-containing proteins. A, there was no significant correlation between the mean largest diameter of tumors and TSP1 expression. B, the largest tumor diameter in the 4N1K-positive group was significantly smaller than that in the 4N1K-negative group. Bars represent the mean largest diameter (cm) of tumor ± 95% confidence interval.
Tables
- Table 1
The relationship between TSP1 expression or 4N1K immunoreactivity and clinicopathological features of RCC
T, N, and M classification were assessed according to the 1997 TNM grading system. Vascular invasion was examined microscopically with the removed tissues.
No. of patients (%) TSP1 expression (%) 4N1K immunoreactivity (%) Negative (n = 70) Positive (n = 49) Negative (n = 87) Positive (n = 32) T classification T1 69 (58.0) 41 (58.6) 28 (57.1) 39 (44.8) 30 (93.8) T2 17 (14.3) 8 (11.4) 9 (18.4) 15 (17.2) 2 (6.2) T3 28 (23.5) 19 (27.1) 9 (18.4) 28 (32.2) 0 (0.0) T4 5 (4.2) 2 (2.9) 3 (6.1) 5 (5.7) 0 (0.0) P 0.448 <0.001 N classification N0 109 (91.6) 63 (90.0) 46 (93.9) 77 (88.5) 32 (100.0) N1 or N2 10 (8.4) 7 (10.0) 3 (4.1) 10 (11.5) 0 (0.0) P 0.453 0.045 M classification M0 102 (85.7) 62 (88.6) 40 (81.6) 70 (80.5) 32 (100.0) M1 17 (14.3) 8 (11.4) 9 (18.4) 17 (19.5) 0 (0.0) P 0.287 0.007 Vascular invasion Negative 89 (74.8) 50 (71.4) 39 (79.6) 60 (69.0) 29 (90.6) Positive 20 (16.8) 20 (29.6) 10 (20.6) 27 (31.0) 3 (9.4) P 0.313 0.016 - Table 2
Ki-67 LI, MVD, and AI of tumor cells in locally confined RCC
To examine the proliferation or apoptosis of tumor cells, sections of tumor tissues were stained with anti-Ki-67 antibody or by the TUNEL method. Ki-67 LI and AI were expressed as positively stained nuclei/total nuclei × 100 (%). For examination of MVD, sections were stained with anti-CD31 antibody to visualize endothelial cells, and MVD was expressed as the number of CD31-positive microvessels/mm2 in tumor tissues. Data represent the mean values ± 95% confidence interval.
TSP1 expression P 4N1K immunoreactivity P Negative (n = 48) Positive (n = 34) Negative (n = 50) Positive (n = 32) Ki-67 LI (%) 5.3 ± 4.9 4.7 ± 3.8 0.249 5.2 ± 4.7 4.8 ± 4.1 0.707 MVD (mm2) 130 ± 55 121 ± 44 0.436 141 ± 51 104 ± 41 <0.001 AI (%) 1.76 ± 0.79 1.85 ± 2.03 0.770 1.51 ± 0.62 2.24 ± 2.10 0.023
Volume 9, Issue 5
