Sphingolipids are ubiquitous components of cell membranes. Their metabolites ceramide, sphingosine, and sphingosine-1-phosphate (S1P) have important physiological functions, including regulation of cell growth and sur...Sphingolipids are ubiquitous components of cell membranes. Their metabolites ceramide, sphingosine, and sphingosine-1-phosphate (S1P) have important physiological functions, including regulation of cell growth and survival. S1P is generated by phosphorylation of sphingosine catalyzed by sphingosine kinase-1 (SPHK1). The purpose of this study is to explore the roles of S1P, S1P receptors, and sphingosine kinases in malignant musculoskeletal tumors. Twenty-one tumor samples (7 liposarcomas, 3 chondrosarcomas, 6 osteosarcomas, 5 MFH) obtained at open biopsy, and four human MFH cell lines (Nara H, Nara F, TNMY1, GBS-1) were used. We examined the mRNA expression of S1P receptors by RT-PCR, and the expression levels of SPHK by Real-time PCR. We used 4 MFH cell lines to analyze SPHK1 proteins by Western blotting. SPHK1 siRNA was transfected into MFH cell lines by lipofection method. Cell proliferation (control and transfected with siRNA) was assayed using WST-8 (Cell Counting Kit-8) assay. All high grade malignant tumors expressed S1P1, S1P2, S1P3 receptors, whereas the expression of S1P1 receptor was detected in 50% of low-grade malignant tumors, S1P2 receptor in 30%, and S1P3 in 50%. No statistically significant difference was found in the expression level of SPHK1 between high-grade and low-grade malignant tumors by Real-time PCR. By results of Western blotting, proteins of SPHK1 were expressed in all MFH cell lines. In MFH cell lines, transfection with SPHK1 siRNA oligonucleotides resulted in approximately 50 to 80% suppression of SPHK1 mRNA expression as determined by real-time PCR. Down-regulation of SPHK1 with small interfering RNA significantly reduced SPHK1 protein levels by Western blotting. Knock down of SPHK1 expression significantly decreased cell proliferation of all MFH cells. These results suggest that the expression of S1P receptors may play an important role for cell proliferation and may correlate with histologic grade in malignant bone and soft tissue tumors, and that SPHK1 may be one of essential molecules for cell proliferation in MFH cell lines.展开更多
This report presents a dedifferentiated chondrosarcoma with a unique pathologic feature. A 63-year-old man was referred with pain and a soft tissue mass in the left groin. A plain radiograph showed a mineralization in...This report presents a dedifferentiated chondrosarcoma with a unique pathologic feature. A 63-year-old man was referred with pain and a soft tissue mass in the left groin. A plain radiograph showed a mineralization in the proximal femur with partially osteolytic foci and an abnormal shadow in the soft tissue. Magnetic resonance imaging scans showed an inhomogeneous lesion with intermediate to partially low signal intensity on T1-weighted image and intermediate to high signal intensity on T2-weighted image. Microscopically, the tumor in the femur is a low-grade chondrosarcoma and the component of soft tissue was a high-grade sarcomatous lesion with an epithelial arrangement of tumor cells. A diffuse immunoreactivity to both vimentin and c-kit (CD117) antibodies was detected in the high-grade component. A dedifferentiated component is similar to those of gastrointestinal stromal tumor (GIST). This is the first case of dedifferentiated chondrosarcoma with a high-grade component mimicking a GIST.展开更多
文摘Sphingolipids are ubiquitous components of cell membranes. Their metabolites ceramide, sphingosine, and sphingosine-1-phosphate (S1P) have important physiological functions, including regulation of cell growth and survival. S1P is generated by phosphorylation of sphingosine catalyzed by sphingosine kinase-1 (SPHK1). The purpose of this study is to explore the roles of S1P, S1P receptors, and sphingosine kinases in malignant musculoskeletal tumors. Twenty-one tumor samples (7 liposarcomas, 3 chondrosarcomas, 6 osteosarcomas, 5 MFH) obtained at open biopsy, and four human MFH cell lines (Nara H, Nara F, TNMY1, GBS-1) were used. We examined the mRNA expression of S1P receptors by RT-PCR, and the expression levels of SPHK by Real-time PCR. We used 4 MFH cell lines to analyze SPHK1 proteins by Western blotting. SPHK1 siRNA was transfected into MFH cell lines by lipofection method. Cell proliferation (control and transfected with siRNA) was assayed using WST-8 (Cell Counting Kit-8) assay. All high grade malignant tumors expressed S1P1, S1P2, S1P3 receptors, whereas the expression of S1P1 receptor was detected in 50% of low-grade malignant tumors, S1P2 receptor in 30%, and S1P3 in 50%. No statistically significant difference was found in the expression level of SPHK1 between high-grade and low-grade malignant tumors by Real-time PCR. By results of Western blotting, proteins of SPHK1 were expressed in all MFH cell lines. In MFH cell lines, transfection with SPHK1 siRNA oligonucleotides resulted in approximately 50 to 80% suppression of SPHK1 mRNA expression as determined by real-time PCR. Down-regulation of SPHK1 with small interfering RNA significantly reduced SPHK1 protein levels by Western blotting. Knock down of SPHK1 expression significantly decreased cell proliferation of all MFH cells. These results suggest that the expression of S1P receptors may play an important role for cell proliferation and may correlate with histologic grade in malignant bone and soft tissue tumors, and that SPHK1 may be one of essential molecules for cell proliferation in MFH cell lines.
文摘This report presents a dedifferentiated chondrosarcoma with a unique pathologic feature. A 63-year-old man was referred with pain and a soft tissue mass in the left groin. A plain radiograph showed a mineralization in the proximal femur with partially osteolytic foci and an abnormal shadow in the soft tissue. Magnetic resonance imaging scans showed an inhomogeneous lesion with intermediate to partially low signal intensity on T1-weighted image and intermediate to high signal intensity on T2-weighted image. Microscopically, the tumor in the femur is a low-grade chondrosarcoma and the component of soft tissue was a high-grade sarcomatous lesion with an epithelial arrangement of tumor cells. A diffuse immunoreactivity to both vimentin and c-kit (CD117) antibodies was detected in the high-grade component. A dedifferentiated component is similar to those of gastrointestinal stromal tumor (GIST). This is the first case of dedifferentiated chondrosarcoma with a high-grade component mimicking a GIST.
