Systemic lupus erythematosus(SLE) and clear cell renal cell carcinoma(CC-RCC) are serious disorders and usually fatal, and always accompanied with pathological changes in the kidney. Signal-induced proliferation-assoc...Systemic lupus erythematosus(SLE) and clear cell renal cell carcinoma(CC-RCC) are serious disorders and usually fatal, and always accompanied with pathological changes in the kidney. Signal-induced proliferation-associated protein 1(SIPA-1) is a Rap1 GTPase activating protein(Rap1GAP) expressed in the normal distal and collecting tubules of the murine kidney. Lupus-like autoimmune disease and leukemia have been observed in SIPA-1 deficient mice, suggesting a pathological relevance of SIPA-1 to SLE and carcinoma in human being. The expression pattern of SIPA-1 is as yet undefined and the pathogenesis of these diseases in humans remains elusive. In this study, we used both immunohistochemistry and quantum dot(QD)-based immunofluorescence staining to investigate the expression of SIPA-1 in renal specimens from SLE and CC-RCC patients. MTT assay and Western blotting were employed to evaluate the effects of SIPA-1 overexpression on the proliferation and apoptosis of renal cell lines. Semi-quantitative reverse transcriptase-PCR(RT-PCR) was applied to examine the changes of hypoxia-inducible factor-1α(HIF-1α) mR NA level. Results showed that SIPA-1 was highly expressed in the proximal and collecting tubules of nephrons in SLE patients compared to normal ones, and similar results were obtained in the specimens of CC-RCC patients. Although SIPA-1 overexpression did not affect cellular proliferation and apoptosis of both human 786-O renal cell carcinoma cells and rat NRK-52 E renal epithelial cell lines, RT-PCR results showed that HIF-1α mR NA level was down-regulated by SIPA-1 overexpression in 786-O cells. These findings suggest that SIPA-1 may play critical roles in the pathological changes in kidney, and might provide a new biomarker to aid in the diagnosis of SLE and CC-RCC.展开更多
To further study the anti metastasis mechanism of laminin glycopeptides on carcinoma cell proliferation, apoptosis and the secretion of matrix metalloproteinases Methods Human hepatocellular carcinoma cells in serum f...To further study the anti metastasis mechanism of laminin glycopeptides on carcinoma cell proliferation, apoptosis and the secretion of matrix metalloproteinases Methods Human hepatocellular carcinoma cells in serum free medium were incubated on laminin coated substrate with or without laminin glycopeptides at a final concentration of 50?μg/ml The total number of surviving cells after incubating for the indicated time was assayed by MTT assay DNA synthesis of the incubated cells was detected by 3H TdR incorporation Cell cycle was analysed by FACS The mitotic index of Giemsa stained cells was assessed Cell apoptosis was detected by both FACS and an acridine orange staining method Matrix metalloproteinase secretion was analysed by gelatin zymography Results The total number of surviving cells incubated on laminin in the absence of laminin glycopeptides was significantly larger than that in the presence of laminin glycopeptides Laminin promoted 3H TdR incorporation of carcinoma cells, decreased the percentage of cells in G1 phase and increased the percentage of cells in S phase In contrast, laminin glycopeptides could inhibit the effect of laminin as shown by 3H TdR incorporation and cell cycle analysis The percentage of cells in G2+M phase and the mitotic index among various groups showed no significant difference Matrix metalloproteinases secretion from cells treated by laminin glycopeptides was much less compared to that without the treatment by laminin glycopeptides Conclusion Laminin may stimulate cell proliferation, while laminin glycopeptides could significantly inhibit the effect of laminin by inhibiting DNA synthesis and arresting the carcinoma cell cycle from G1 to S phase These effects may inhibit not only tumor growth of the primary carcinoma, but also the establishment of metastases at ectopic tissues Laminin glycopeptides could also inhibit the secretion of matrix metalloproteinases from carcinoma cells and this may contribute to their decreased invasive and metastatic phenotype This study further revealed the cellular and molecular mechanism of laminin glycopeptides on anti展开更多
基金supported by grants from National Natural Science Foundation of China(No.31271504)Shenzhen Science and Technology Innovations Committee,China(No.JCYJ20130401144744187)Health Department of Hubei Province,China(No.JX6A06)
文摘Systemic lupus erythematosus(SLE) and clear cell renal cell carcinoma(CC-RCC) are serious disorders and usually fatal, and always accompanied with pathological changes in the kidney. Signal-induced proliferation-associated protein 1(SIPA-1) is a Rap1 GTPase activating protein(Rap1GAP) expressed in the normal distal and collecting tubules of the murine kidney. Lupus-like autoimmune disease and leukemia have been observed in SIPA-1 deficient mice, suggesting a pathological relevance of SIPA-1 to SLE and carcinoma in human being. The expression pattern of SIPA-1 is as yet undefined and the pathogenesis of these diseases in humans remains elusive. In this study, we used both immunohistochemistry and quantum dot(QD)-based immunofluorescence staining to investigate the expression of SIPA-1 in renal specimens from SLE and CC-RCC patients. MTT assay and Western blotting were employed to evaluate the effects of SIPA-1 overexpression on the proliferation and apoptosis of renal cell lines. Semi-quantitative reverse transcriptase-PCR(RT-PCR) was applied to examine the changes of hypoxia-inducible factor-1α(HIF-1α) mR NA level. Results showed that SIPA-1 was highly expressed in the proximal and collecting tubules of nephrons in SLE patients compared to normal ones, and similar results were obtained in the specimens of CC-RCC patients. Although SIPA-1 overexpression did not affect cellular proliferation and apoptosis of both human 786-O renal cell carcinoma cells and rat NRK-52 E renal epithelial cell lines, RT-PCR results showed that HIF-1α mR NA level was down-regulated by SIPA-1 overexpression in 786-O cells. These findings suggest that SIPA-1 may play critical roles in the pathological changes in kidney, and might provide a new biomarker to aid in the diagnosis of SLE and CC-RCC.
文摘To further study the anti metastasis mechanism of laminin glycopeptides on carcinoma cell proliferation, apoptosis and the secretion of matrix metalloproteinases Methods Human hepatocellular carcinoma cells in serum free medium were incubated on laminin coated substrate with or without laminin glycopeptides at a final concentration of 50?μg/ml The total number of surviving cells after incubating for the indicated time was assayed by MTT assay DNA synthesis of the incubated cells was detected by 3H TdR incorporation Cell cycle was analysed by FACS The mitotic index of Giemsa stained cells was assessed Cell apoptosis was detected by both FACS and an acridine orange staining method Matrix metalloproteinase secretion was analysed by gelatin zymography Results The total number of surviving cells incubated on laminin in the absence of laminin glycopeptides was significantly larger than that in the presence of laminin glycopeptides Laminin promoted 3H TdR incorporation of carcinoma cells, decreased the percentage of cells in G1 phase and increased the percentage of cells in S phase In contrast, laminin glycopeptides could inhibit the effect of laminin as shown by 3H TdR incorporation and cell cycle analysis The percentage of cells in G2+M phase and the mitotic index among various groups showed no significant difference Matrix metalloproteinases secretion from cells treated by laminin glycopeptides was much less compared to that without the treatment by laminin glycopeptides Conclusion Laminin may stimulate cell proliferation, while laminin glycopeptides could significantly inhibit the effect of laminin by inhibiting DNA synthesis and arresting the carcinoma cell cycle from G1 to S phase These effects may inhibit not only tumor growth of the primary carcinoma, but also the establishment of metastases at ectopic tissues Laminin glycopeptides could also inhibit the secretion of matrix metalloproteinases from carcinoma cells and this may contribute to their decreased invasive and metastatic phenotype This study further revealed the cellular and molecular mechanism of laminin glycopeptides on anti
