Objective： To detect the expression of KISS-1 and S100A4 in the primary tumor tissues and lymphatic and visceral metastases and investigate its role in tumorigenesis and metastasis of gastric cancer. Methods： The pr...Objective： To detect the expression of KISS-1 and S100A4 in the primary tumor tissues and lymphatic and visceral metastases and investigate its role in tumorigenesis and metastasis of gastric cancer. Methods： The protein expression of KISS-1 and S100A4 in lymphatic and visceral metastases from advanced gastric cancer specimens was mainly examined by immunohistochemical staining and tissues microarray. Results： Immunohistochemical staining revealed reduced expression of KISS-1 and up-regulated expression of S100A4 in lymph node and visceral metastases. Rates of KISS-1 expression in normal tissues, primary tumor tissues, lymph node and visceral metastases were 95.8%, 74.6%, 60.9% and 57.5%. $100A4 expression in associated cases was 43.6%, 71.8%, 70.3% and 90.0%, respectively. Significant differences in KISS-1 expression was significantly higher in normal tissues than that in primary tumor tissues （P〈0.001）. While significant differences of S100A4 expression could be seen between normal and cancer tissues （P〈0.001） and between visceral and primary tumors （P〈0.05）. Conclusion： Tumor metastasis results from gradual accumulation of abnormal genetic alterations. Down-regulation of KISS-1 and up-regulation of S100A4 play a critical role in metastasis of gastric carcinoma.展开更多
Objective： The aim of the study was to investigate inhibitory effects of breast-cancer metastasis suppressor 1 protein （BRMS1） on primary tumor growth and metastasis of human gastric cancer （GC） cells in nude mic...Objective： The aim of the study was to investigate inhibitory effects of breast-cancer metastasis suppressor 1 protein （BRMS1） on primary tumor growth and metastasis of human gastric cancer （GC） cells in nude mice. Methods： We compared the expression of BRMS1 in the primary gastric tumor and metastatic gastric tumor by immunohistochemistry. Expression of BRMS1 also was detected in the GC cells by RT-PCR and Western blot. Three groups of cultured human GC cell line SGC-7901, were maintained： transfected cells with pcDNA3.1（-）B/myc-BRMS1; negative control cells with pcD- NA3.1/myc-his（-）B; and blank control ceils without any transfection. Histologically intact samples of the cells, maintained by passage in the subcutis of nude mice, were transplanted orthotopically into stomach walls of nude mice to establish a nude mouse model of human gastric carcinoma. Their primary tumor growth and metastasis were then observed. Results： The expression of BRMS1 was markedly stronger in the primary gastric tumor compared with metastatic gastric tumor. We also detected BRMS1 gene and protein in the gastric cancer cell tines. Numbers of metastasis tumors significantly differed among mice infected with transfected cells, with negative controls and with blank controls （4.38 ± 0.60, 7.75 ± 0.59, and 7.63± 0.65, respectively; P 〈 0.05）. However, there were no significant differences in the size of orthotopic tumors among mice infected with transfected, negative control and blank control cells [（12.02 ± 0.70）, （12.71 ± 0.63） and （12.89 ± 0.71） mm, respectively; P 〉 0.05]. Conclusion： BRMS1 suppresses metastasis of GC cells, but does not inhibit growth of gastric tumors.展开更多
AIM:To investigate the function of the KISS-1 gene in gastric carcinoma cells and to explore its potential mechanism.METHODS:A KISS-1 eukaryotic expression vector was constructed and transfected into BGC-823 cells.Res...AIM:To investigate the function of the KISS-1 gene in gastric carcinoma cells and to explore its potential mechanism.METHODS:A KISS-1 eukaryotic expression vector was constructed and transfected into BGC-823 cells.Resistant clones were obtained through G418 selection.reverse transcription-polymerase chain reaction and western blotting were used to detect KISS-1 and matrix metalloproteinase-9(MMP-9)expression in transfected cells.The growth of transfected cells was investigated by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT)proliferation assays,and the cells'invasive potential was analyzed by basement membrane(Matrigel)invasion assays.The anti-tumor effects of KISS-1 were tested in vivo using allografts in nude mice.RESULTS:The expression level of KISS-1 mRNA andprotein in BGC-823/KISS-1 transfected cells were significantly higher than in BGC-823/pcDNA3.1 transfected cells(P<0.05)or the parental BGC-823 cell line(P< 0.05).The expression level of MMP-9 mRNA and protein in BGC-823/KISS-1 were significantly less than in BGC-823/pcDNA3.1(P<0.05)or BGC-823 cells(P< 0.05).MTT growth assays show the proliferation of BGC-823/KISS-1 cells at 48 h(0.642±0.130)and 72 h(0.530±0.164)were significantly reduced compared to BGC-823/pcDNA3.1(0.750±0.163,0.645±0.140)(P<0.05)and BGC-823 cells(0.782±0.137,0.685± 0.111)(P<0.05).Invasion assays indicate the invasive potential of BGC-823/KISS-1 cells(16.50±14.88)is significantly reduced compared to BGC-823/pcDNA3.1(20.22±14.87)(P<0.05)and BGC-823 cells after 24 h(22.12±16.12)(P<0.05).In vivo studies demonstrate the rate of pcDNA3.1-KISS-1 tumor growth is significantly slower than pcDNA3.1 and control cell tumor growth in nude mice.Furthermore,tumor volume of pcDNA3.1-KISS-1 tumors(939.38±82.08 mm3)was significantly less than pcDNA3.1(1250.46±44.36 mm3) and control tumors(1284.36±55.26 mm3)(P<0.05).Moreover,the tumor mass of pcDNA3.1-KISS-1 tumors(0.494±0.84 g)was significantly less than pcDNA3.1(0.668±0.55 g)and control tumors(0.682±0.38 g)(P <0.05).CONCLUSION:KISS-1 may inhibit the proliferation and invasion of gastric carcinoma cells in vitro and in vivo through the downregulation of MMP-9.展开更多
Objective： The aim of this study was to establish the osteosarcoma cell sublines which stably expressing tumor suppressor in lung cancer-1 （TSLC1） gene and evaluate its effect on growth inhibition of human osteosar...Objective： The aim of this study was to establish the osteosarcoma cell sublines which stably expressing tumor suppressor in lung cancer-1 （TSLC1） gene and evaluate its effect on growth inhibition of human osteosarcoma cell line MG63. Methods： The recombinant plasmid pCI-TSLC1 was stably transfected into MG63 cells with Lipofectamine 2000. The posi- tive clones were developed by selection by G418. Biological characteristics of one of the 6 cell lines which highly expressing TSLC1, namely, the M8T were studied. Cell growth was analyzed with MTT assay. 2 x 10^7cells suspended in 0.2 mL phosphate buffered saline （PBS） were injected into the two flanks of 5-6-week-old female BALB/C nu/nu athymic nude mice. The volumes of subcutaneous of tumor growth were evaluated and calculated by the formula V= Length x Width x Height x 0.5 once a week. Results： The MST cell subline which stably expressing TSLC1 was characterized by Western blot. The genetic stability and purity of M8T cells were stable. TSLC1 significantly suppressed the growth of M8T cells in vitro. Moreover, the tumorigenicity of MST cells was suppressed in vivo. Conclusion： The osteosarcoma cell sublines MST which stably expressing TSLC1 had been successfully established. The ability of growth and metastasis of MST was significantly suppressed both in vitro and in vivo.展开更多
文摘Objective： To detect the expression of KISS-1 and S100A4 in the primary tumor tissues and lymphatic and visceral metastases and investigate its role in tumorigenesis and metastasis of gastric cancer. Methods： The protein expression of KISS-1 and S100A4 in lymphatic and visceral metastases from advanced gastric cancer specimens was mainly examined by immunohistochemical staining and tissues microarray. Results： Immunohistochemical staining revealed reduced expression of KISS-1 and up-regulated expression of S100A4 in lymph node and visceral metastases. Rates of KISS-1 expression in normal tissues, primary tumor tissues, lymph node and visceral metastases were 95.8%, 74.6%, 60.9% and 57.5%. $100A4 expression in associated cases was 43.6%, 71.8%, 70.3% and 90.0%, respectively. Significant differences in KISS-1 expression was significantly higher in normal tissues than that in primary tumor tissues （P〈0.001）. While significant differences of S100A4 expression could be seen between normal and cancer tissues （P〈0.001） and between visceral and primary tumors （P〈0.05）. Conclusion： Tumor metastasis results from gradual accumulation of abnormal genetic alterations. Down-regulation of KISS-1 and up-regulation of S100A4 play a critical role in metastasis of gastric carcinoma.
基金Supported by a grant from the Jiangxi Provincial Natural Science Foundation of China (No. 0640063)
文摘Objective： The aim of the study was to investigate inhibitory effects of breast-cancer metastasis suppressor 1 protein （BRMS1） on primary tumor growth and metastasis of human gastric cancer （GC） cells in nude mice. Methods： We compared the expression of BRMS1 in the primary gastric tumor and metastatic gastric tumor by immunohistochemistry. Expression of BRMS1 also was detected in the GC cells by RT-PCR and Western blot. Three groups of cultured human GC cell line SGC-7901, were maintained： transfected cells with pcDNA3.1（-）B/myc-BRMS1; negative control cells with pcD- NA3.1/myc-his（-）B; and blank control ceils without any transfection. Histologically intact samples of the cells, maintained by passage in the subcutis of nude mice, were transplanted orthotopically into stomach walls of nude mice to establish a nude mouse model of human gastric carcinoma. Their primary tumor growth and metastasis were then observed. Results： The expression of BRMS1 was markedly stronger in the primary gastric tumor compared with metastatic gastric tumor. We also detected BRMS1 gene and protein in the gastric cancer cell tines. Numbers of metastasis tumors significantly differed among mice infected with transfected cells, with negative controls and with blank controls （4.38 ± 0.60, 7.75 ± 0.59, and 7.63± 0.65, respectively; P 〈 0.05）. However, there were no significant differences in the size of orthotopic tumors among mice infected with transfected, negative control and blank control cells [（12.02 ± 0.70）, （12.71 ± 0.63） and （12.89 ± 0.71） mm, respectively; P 〉 0.05]. Conclusion： BRMS1 suppresses metastasis of GC cells, but does not inhibit growth of gastric tumors.
基金Supported by The Natural Science Foundation,No.2011A310005the Science and Technique Foundation of Henan Province,No.112102310206
文摘AIM:To investigate the function of the KISS-1 gene in gastric carcinoma cells and to explore its potential mechanism.METHODS:A KISS-1 eukaryotic expression vector was constructed and transfected into BGC-823 cells.Resistant clones were obtained through G418 selection.reverse transcription-polymerase chain reaction and western blotting were used to detect KISS-1 and matrix metalloproteinase-9(MMP-9)expression in transfected cells.The growth of transfected cells was investigated by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT)proliferation assays,and the cells'invasive potential was analyzed by basement membrane(Matrigel)invasion assays.The anti-tumor effects of KISS-1 were tested in vivo using allografts in nude mice.RESULTS:The expression level of KISS-1 mRNA andprotein in BGC-823/KISS-1 transfected cells were significantly higher than in BGC-823/pcDNA3.1 transfected cells(P<0.05)or the parental BGC-823 cell line(P< 0.05).The expression level of MMP-9 mRNA and protein in BGC-823/KISS-1 were significantly less than in BGC-823/pcDNA3.1(P<0.05)or BGC-823 cells(P< 0.05).MTT growth assays show the proliferation of BGC-823/KISS-1 cells at 48 h(0.642±0.130)and 72 h(0.530±0.164)were significantly reduced compared to BGC-823/pcDNA3.1(0.750±0.163,0.645±0.140)(P<0.05)and BGC-823 cells(0.782±0.137,0.685± 0.111)(P<0.05).Invasion assays indicate the invasive potential of BGC-823/KISS-1 cells(16.50±14.88)is significantly reduced compared to BGC-823/pcDNA3.1(20.22±14.87)(P<0.05)and BGC-823 cells after 24 h(22.12±16.12)(P<0.05).In vivo studies demonstrate the rate of pcDNA3.1-KISS-1 tumor growth is significantly slower than pcDNA3.1 and control cell tumor growth in nude mice.Furthermore,tumor volume of pcDNA3.1-KISS-1 tumors(939.38±82.08 mm3)was significantly less than pcDNA3.1(1250.46±44.36 mm3) and control tumors(1284.36±55.26 mm3)(P<0.05).Moreover,the tumor mass of pcDNA3.1-KISS-1 tumors(0.494±0.84 g)was significantly less than pcDNA3.1(0.668±0.55 g)and control tumors(0.682±0.38 g)(P <0.05).CONCLUSION:KISS-1 may inhibit the proliferation and invasion of gastric carcinoma cells in vitro and in vivo through the downregulation of MMP-9.
基金Supported by a grant from the National Natural Science Foundation of Hubei Province (No. 2010CDB09302)
文摘Objective： The aim of this study was to establish the osteosarcoma cell sublines which stably expressing tumor suppressor in lung cancer-1 （TSLC1） gene and evaluate its effect on growth inhibition of human osteosarcoma cell line MG63. Methods： The recombinant plasmid pCI-TSLC1 was stably transfected into MG63 cells with Lipofectamine 2000. The posi- tive clones were developed by selection by G418. Biological characteristics of one of the 6 cell lines which highly expressing TSLC1, namely, the M8T were studied. Cell growth was analyzed with MTT assay. 2 x 10^7cells suspended in 0.2 mL phosphate buffered saline （PBS） were injected into the two flanks of 5-6-week-old female BALB/C nu/nu athymic nude mice. The volumes of subcutaneous of tumor growth were evaluated and calculated by the formula V= Length x Width x Height x 0.5 once a week. Results： The MST cell subline which stably expressing TSLC1 was characterized by Western blot. The genetic stability and purity of M8T cells were stable. TSLC1 significantly suppressed the growth of M8T cells in vitro. Moreover, the tumorigenicity of MST cells was suppressed in vivo. Conclusion： The osteosarcoma cell sublines MST which stably expressing TSLC1 had been successfully established. The ability of growth and metastasis of MST was significantly suppressed both in vitro and in vivo.
