PMC3616025_01
The breast mass involving almost the whole breast with ulcerated nipple.
amjcaserep-13-218-g001.jpg
PMC3616025_01_amjcaserep-13-218-g001.jpg
f1-amjcaserep-13-218
The mass was associated with bilateral axillary lymph nodes enlargement, hard and amalgamated on the ipsilateral side, while mobile and firm on the contralateral one (Figure 1).
(A) The tumor area from the breast mass showing invasive groups of malignant moderately differentiated ductal cells (x), (B) the same tumor area showing positive immunostaining of the malignant ductal cells for Her 2 neu (+3) (x200), (C) a microscopic picture of the bony lesion showing malignant spindle cells associated with deposition of osteoid, which supports the diagnosis of osteosarcoma, and (D) negative immunostaining for cytokeratin CK in the malignant osteoblasts.
amjcaserep-13-218-g002.jpg
PMC3616025_01_amjcaserep-13-218-g002.jpg
f2-amjcaserep-13-218
II with scattered lympho-vascular permeation (Figure 2A,B). Immunostaining for estrogen and progesterone receptors were positive (ER+, PR +), and Her-2/neu was overexpressed (score 3). Pathological examination showed a malignant neoplasm formed of proliferating pleomorphic large cells having scanty cytoplasm, and hyperchromatic nuclei with related deposition of thin lace-like branching osteoid matrix and negative immunostaining for CK (Figure 2C,D), excluding breast origin and confirming the diagnosis of conventional osteosarcoma GII, osteoblastic type.
X-ray of the left femur showing an ill defined irregular osteoscelorotic lesion in the supracondylar region with soft tissue extension.
amjcaserep-13-218-g003.jpg
PMC3616025_01_amjcaserep-13-218-g003.jpg
f3-amjcaserep-13-218
Local X-ray of the left femur showed an irregular, dense, ill-defined, osteosclerotic lesion, with possible soft tissue extension (Figure 3), which was confirmed by MRI.
Sequence analysis of the p53 gene for the two lesions showing mutation in the p53 gene (GCC to ACC) in the osteosarcoma (OS) but not in breast invasive duct carcinoma (IDC).
amjcaserep-13-218-g004.jpg
PMC3616025_01_amjcaserep-13-218-g004.jpg
f4-amjcaserep-13-218
BT and OS, though at different codons (Figure 4), while K13-ras mutations was detected in OS only (Figure 5). BRCA1 gene mutations were concentrated at exons 11 and 1, whereas LOH concentrated at exons 11 followed by 6 (Tables 1, 2).
An ethedium bromide-stained gel showing the wild type of codon 12-ras (125bp band) both in OS and IDC. Mutation in codon 13-ras was detected in OS (157bp band) only whereas the IDC showed no mutations.
amjcaserep-13-218-g005.jpg
PMC3616025_01_amjcaserep-13-218-g005.jpg
f5-amjcaserep-13-218
BT and OS, though at different codons (Figure 4), while K13-ras mutations was detected in OS only (Figure 5). BRCA1 gene mutations were concentrated at exons 11 and 1, whereas LOH concentrated at exons 11 followed by 6 (Tables 1, 2).
An ethedium bromide-stained gel for the RT-PCR showing reduction/loss of BRCA1, BRCA2, MLH1, MSH2 mRNA in the IDC as well as in the OS samples. BRCA2 showed reduced expression in OS only while MSH6 was normally expressed.
amjcaserep-13-218-g006.jpg
PMC3616025_01_amjcaserep-13-218-g006.jpg
f6-amjcaserep-13-218
No mutations were detected in BRCA2 gene, though reduced mRNA expression of BRCA1&2 (Figure 6) was evident in both tumor samples.
Methylation specific PCR showing promoter hypermethylation (PM) of MLH1 and MSH2 genes both in the IDC and OS samples.
amjcaserep-13-218-g007.jpg
PMC3616025_01_amjcaserep-13-218-g007.jpg
f7-amjcaserep-13-218
MLH1, MSH2 mRNA expression was reduced in both tumors, especially in the OS, whereas MSH6 was normally expressed and RRM1 was overexpressed in both tumors. MLH1 and MSH2 showed PM in both tumors (Figure 7), MSI in BT, and different mutational events in both tumors (Tables 1, 2 & Figure 8).
Microsatellite instability (MSI) from the IDC and the OS showing MSI in the MLH1 and MSH2 gene loci in the IDC but not in OS.
amjcaserep-13-218-g008.jpg
PMC3616025_01_amjcaserep-13-218-g008.jpg
f8-amjcaserep-13-218
MLH1, MSH2 mRNA expression was reduced in both tumors, especially in the OS, whereas MSH6 was normally expressed and RRM1 was overexpressed in both tumors. MLH1 and MSH2 showed PM in both tumors (Figure 7), MSI in BT, and different mutational events in both tumors (Tables 1, 2 & Figure 8).