Article Figures & Data
Figures
- Fig. 1.
Screening of autoantibodies in breast cancer. A, proteome profile of SUM-44 cells. The proteins from whole-cell extracts of SUM-44 cells were separated by 2-D PAGE and silver stained. B, close up sections showing Western blot performed with lysates from SUM-44 cells and sera from healthy controls or from breast cancer patients as primary antibody. An antihuman IgG was used as a secondary antibody. The three spots recognized by sera from patients with breast cancer are indicated by open arrows.
- Fig. 2.
Characterization of RS by Western blot. The proteins from the breast cell line SUM-44 were separated by 2-D PAGE. A, the proteins were transferred to a PVDF membrane, and Western blotting was performed using the rabbit polyclonal antibody directed against RS or the mouse monoclonal antibody directed against DJ-1. B, the proteins from SUM-44 cells were silver stained. The five different forms of RS/DJ-1 are labeled. Only a part of the gel is shown.
- Fig. 3.
RS/DJ-1 expression analysis by Western blot. A, the proteins from several cell lines were separated by 2-D PAGE and transferred to a PVDF membrane. The expression of RS/DJ-1 was investigated by Western blotting using the rabbit polyclonal antibody directed against RS and whole-cell extracts from the cell lines SUM-44 (Breast), HuH7 (Liver), A549 (Lung), and U87 (Brain). B, RS/DJ-1 expression was investigated in normal mammary epithelium as well as in three primary breast tumors.
- Fig. 4.
IHC. Sections of breast tissue containing healthy and tumor cell areas were immunostained with the rabbit polyclonal antibody against RS. The nuclei in the epithelial cells from the healthy tissue were stained with a high intensity, whereas most of nuclei in the carcinoma area were not stained. These results are representative of the observations for 20 slides.
- Fig. 5.
Occurrence of RS/DJ-1 in the secreted protein fraction of SUM-44. Proteins from whole cell extracts (WCE; left) or from supernatant of SUM-44 cells (SN; right) were separated by 2-D PAGE and transferred to a PVDF membrane. Expression of RS/DJ-1 and β-tubulin was determined by Western blotting experiments.
- Fig. 6.
Presence of the protein RS/DJ-1 in the sera from patients with breast cancer. A, serum proteins from breast cancer patients or from healthy women were separated by 1-D PAGE and transferred to a PVDF membrane. Western blotting experiments were performed with the mouse monoclonal antibody against DJ-1. B, serum containing the protein RS/DJ-1 were used for Western blotting experiments after 2-D PAGE. The different RS/DJ-1 isoforms of the protein RS/DJ-1 are circulating.
Tables
- Table 1
Clinical characteristics of subjects with breast cancer
Patient Age (yrs) Stage Pathology RS Aba RS Ag− 1 38 T2N1M0 Infiltrat. ductal Ab+ Ag+ 2 50 T1N0M0 DI Ab− Ag− 3 54 T2N0M0 DI Ab− Ag− 4 57 T3N1bM0 DI Ab− Ag− 5 84 T2N0M0 Mucinous Ab− Ag+ 6 72 T2N0M0 DI Ab− Ag− 7 52 T1N0M0 DI Ab− Ag− 8 78 T4dN3M0 DI Ab− Ag− 9 86 T3N0M0 DI Ab− Ag+ 10 37 T1N0M0 Infiltrat. lobula Ab− Ag− 11 72 T2N0M0 DI Ab− Ag− 12 47 T2N0M0 DI Ab+ Ag+ 13 72 T1N0M0 Infiltrat. lobula Ab− Ag− 14 72 T2N0M0 DI Ab− Ag− 15 42 T2N1M1 DI Ab− Ag− 16 49 T1N0M0 DI Ab− Ag+ 17 62 Recurrence Infiltrat. lobula Ab− Ag− 18 66 T0N0M0 Intraductal Ab− Ag+ 19 65 Preinvasive Intraductal Ab− Ag+ 20 56 T4N1aM1 DI Ab− Ag− 21 46 T1N0 DI Ab− Ag+ 22 55 T1N0M0 DI Ab− Ag+ 23 45 T2N0 DI Ab− Ag− 24 65 T3N0M3 DI Ab− Ag− 25 64 T3N1aM0 DI Ab+ Ag+ 26 63 T2N0M0 DI Ab− Ag− 27 77 T2N0 DI Ab− Ag− 28 87 T2NXM0 DI Ab− Ag− 29 75 DI Ab+ Ag+ 30 40 T1N0M0 DI Ab− Ag− -
a Ab, antibody; Ag, antigen; DI, Infiltrat. ductal.
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- Table 2
Assignment of peptide masses to the human RS/DJ-1 sequence
Analysis by MALDI-TOF mass spectrometry of the peptide masses followed by search in the NCBI database. Measured mass (Da) Matched (Da) Delta (ppm) Residues Sequence Trypsin 875.8823 875.9826 114.5041 42–48 DPVQCSR 1159.3799 1159.2890 78.3719 90 –98 EILKEQENR 1547.0638 1546.7425 207.7298 49 –62 DVVICPDASLEDAK 1677.4343 1676.6554 285.6045 13 –27 GAEEMETVIPVDVMR 2283.2122 2282.7201 215.5653 100–122 GLIAAICAGPTALLAHEIGFGSK 2410.9219 2410.8949 11.1956 99–122 KGLIAAICAGPTALLAHEIGFGSK 2385.8840 2835.9015 −7.3287 6–27 ALVILAKGAEEMETVIPVDVMR 2586.1042 2585.8833 85.4138 64–89 EGPYDVVVLPGGNLGAQNLSESAAVK Chymotrypsin 983.4457 983.1567 293.9127 68 –77 DVVVLPGGNL 1013.2704 1013.2734 −3.0080 1–10 ASKRALVILa 1072.0384 1072.3168 −259.6541 102 –112 IAAICAGPTAL 1146.9828 1147.2344 −219.3025 142 –151 SENRVEKDGL 1186.4784 1186.2706 175.1733 154 –164 TSRGPGTSFEF 2942.1274 2942.5262 −135.5434 11–38 AKGAEEMETVIPVDVMRRAGIKVTVAGL 3235.2369 3234.6441 183.2789 39–67 AGKDPVQCSRDVVICPDASLEDAKKEGPY Glu-C 760.9006 760.8281 95.2907 144 –149 NRVEKD 1117.1193 1117.3111 −171.6654 50 –59 VVICPDASLE 1352.8936 1352.6664 168.0011 177 –189 VAAQVKAPLVLKD 1375.4694 1375.6134 −104.7112 164–176 FALAIVEALNGKE 1435.4879 1435.6256 −95.9287 150–163 GLILTSRGPGTSFE 1567.5786 1567.7887 −134.0037 69–84 VVVLPGGNLGAQNLSE 1598.6701 1598.8685 −124.0827 117–131 IGCGSKVTTHPLAKD 1598.84.02 1598.8900 −31.1375 1–16 ASKRALVILAKGAEEa 1678.6752 1678.8892 −127.4453 148–163 KDGLILTSRGPGTSFE 2971.5911 2971.3955 65.8432 117–143 IGFGSKVTTHPLAKDKMMNGGHYTYSE 2972.2967 2971.8563 171.2222 60–84 DAKKEGPYDVVVLPGGNLGAQNLSE -
a The peptide mass match is compatible with an Acet-N modification.
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Volume 7, Issue 11
