AIM: To study the expression of early growth response gene1 (Egr-1 gene) and Bcl-X/L protein and its relationship with the cell apoptosis in human esophageal carcinoma(EC) and precancerous lesions.METHODS: In situ hyb...AIM: To study the expression of early growth response gene1 (Egr-1 gene) and Bcl-X/L protein and its relationship with the cell apoptosis in human esophageal carcinoma(EC) and precancerous lesions.METHODS: In situ hybridization(ISH), immunohistochemistry (IHC) and TUNEL method were used respectively to detect Egr-1mRNA, Egr-1 protein, apoptosis related-protein Bcl-X/L and cell apoptosis in situ from 66 cases of esophageal squamous cell carcinoma and their upper cut edge and paracancerous mucosa.RESULTS: Egr-1 gene in situ hybridization, Bcl-X/L immunohistochemistry positive products were located in the cytoplasm, while Egr-1 immunohistochemistry and TUNEL positive signal were located in the nuclei. The apoptosis index(AI) and the frequency of apoptosis occurrence were increased gradually from precancerous lesion to cancer (P＜0.01) and the expression of Egr-1mRNA and Egr-1 protein in dysplasia was the highest among all specimens (P＜0.01).The AI of Egr-1 positive cancer tissues was much higher than that of Egr-1 negative cancer tissues (P＜0.01), while the AI of Bcl-X/L positive cancer tissues was much lower than that of Bcl-X/L negative cancer tissues (P＜0.01). The AI and Egr-1 expression were not correlated with invasiveness and lymphatic metastasis in EC.CONCLUSION: Cell apoptosis was present through esophageal carcinogenesis. The expression of Egr-1 mRNA and Egr-1 protein were high in precancerous lesion of esophagus. The AI was increased significantly in Egr-1 positive squamous cell carcinoma. Egr-1 might promote apoptotic effect. Egr-1 expression and cell apoptosis may have an important biological significance in esophageal carcinogenesis.展开更多
AIM:To examine the expression of Egr-1, c-fos and cyclin D1 at both transcript and protein levels in esophageal carcinoma and to correlate the level of their expressions with precancerous and paracancerous esophageal ...AIM:To examine the expression of Egr-1, c-fos and cyclin D1 at both transcript and protein levels in esophageal carcinoma and to correlate the level of their expressions with precancerous and paracancerous esophageal lesions and esophageal carcinoma.METHODS:In situ hybridization and immunohistochemistry were used respectively to detect the expression of mRNA and proteins of Egr-1, c-fos and cyclin D1 in 70 cases of esophageal squamous cell carcinoma and their corresponding para-cancerous mucosa and upper cut edge mucosa.RESULTS:In situ hybridization and immunohistochemistry showed positive staining of all three mRNAs in the cytoplasm and those of the proteins in nuclei. Overexpression of Egr-1, c-los and cyclin D1 mRNAs and their proteins was found in dysplasia and squamous carcinomas. The expression level of Egr-1 and c-los was high, and cyclin D1 was low in dysplasia mucosa, whereas the expression of Egr-1 was decreased, c-fos was maintained and cyclin D1 was increased in the cancers. The expression of both c-fos and cyclinD1 was consistent between the mRNA and protein in their corresponding high expression lesions.CONCLUSION: The expression of Egr-1, c-fos and cyclin D1 varies in esophageal precancerous lesions and cancer tissues, suggesting an involvement of these genes in the development of esophageal carcinoma.展开更多
This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellu-lar growth, apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro. The expression of TRAIL and en...This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellu-lar growth, apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro. The expression of TRAIL and endostatin protein in ECV304 cells was detected by ELISA after the transfection of recombinant plasmid pshuttle-Egr1-shTRAIL-shES and X-ray irradiation. Then MTT assay was used for determining the cellular proliferation, and flow cytometry (FCM) plus Annexin V and propidium iodide (PI) double-staining or PI single-staining were employed for the detection of apoptosis and cell cycle progression. The results showed that expression of TRAIL and endostatin protein exhibited a time- and dose-dependent change in ECV304 cells after pshut-tle-Egr1-shTRAIL-shES transfection in conjunction with irradiation. In the TRAIL-endostatin-based single- or double-gene-radiotherapy, the cell viability declined in a time- and dose-dependent manner, the percentage of cells at G2/M phase and apoptotic rate was increased, and the percentage of cells at G0/G1 phase was lowered as compared with those receiving radiotherapy alone. Moreover, TRAIL-endostatin-based double-gene-radiotherapy demonstrated better effects on growth inhibition, promotion of apoptosis and induction of cell cycle arrest in ECV304 cells than single-gene-radiotherapy.展开更多
目的 :克隆小鼠内皮抑素 (m Endostatin)编码区 c DNA序列并构建含 Egr- 1启动子的 IFNγ和 m Endo-statin双基因表达载体。方法 :利用逆转录多聚酶链反应法 (RT- PCR) ,以小鼠肝细胞 m RNA为模板 ,扩增获得全长 m Endostatin,与 p MD1...目的 :克隆小鼠内皮抑素 (m Endostatin)编码区 c DNA序列并构建含 Egr- 1启动子的 IFNγ和 m Endo-statin双基因表达载体。方法 :利用逆转录多聚酶链反应法 (RT- PCR) ,以小鼠肝细胞 m RNA为模板 ,扩增获得全长 m Endostatin,与 p MD1 8T载体连接作全自动测序 ,并利用基因重组技术构建含 Egr- 1启动子的 IFNγ和m Endostatin双基因表达质粒。结果 :经测序证实获得的 m Endostatin序列与文献报道完全一致 ,并构建了含 Egr-1启动子的 IFNγ和 m Endostatin双基因表达质粒 p Egr- IFNγ- m Endostatin。结论 :利用 RT- PCR法成功克隆了m Endostatin的 c DNA序列 ,构建了 p Egr- IFNγ- m Endostatin重组双基因表达质粒。展开更多
基金the National Natural Science Foundation of China,No.39670298
文摘AIM: To study the expression of early growth response gene1 (Egr-1 gene) and Bcl-X/L protein and its relationship with the cell apoptosis in human esophageal carcinoma(EC) and precancerous lesions.METHODS: In situ hybridization(ISH), immunohistochemistry (IHC) and TUNEL method were used respectively to detect Egr-1mRNA, Egr-1 protein, apoptosis related-protein Bcl-X/L and cell apoptosis in situ from 66 cases of esophageal squamous cell carcinoma and their upper cut edge and paracancerous mucosa.RESULTS: Egr-1 gene in situ hybridization, Bcl-X/L immunohistochemistry positive products were located in the cytoplasm, while Egr-1 immunohistochemistry and TUNEL positive signal were located in the nuclei. The apoptosis index(AI) and the frequency of apoptosis occurrence were increased gradually from precancerous lesion to cancer (P＜0.01) and the expression of Egr-1mRNA and Egr-1 protein in dysplasia was the highest among all specimens (P＜0.01).The AI of Egr-1 positive cancer tissues was much higher than that of Egr-1 negative cancer tissues (P＜0.01), while the AI of Bcl-X/L positive cancer tissues was much lower than that of Bcl-X/L negative cancer tissues (P＜0.01). The AI and Egr-1 expression were not correlated with invasiveness and lymphatic metastasis in EC.CONCLUSION: Cell apoptosis was present through esophageal carcinogenesis. The expression of Egr-1 mRNA and Egr-1 protein were high in precancerous lesion of esophagus. The AI was increased significantly in Egr-1 positive squamous cell carcinoma. Egr-1 might promote apoptotic effect. Egr-1 expression and cell apoptosis may have an important biological significance in esophageal carcinogenesis.
基金Supported by the National Natural Science Foundation of China,No.39670298
文摘AIM:To examine the expression of Egr-1, c-fos and cyclin D1 at both transcript and protein levels in esophageal carcinoma and to correlate the level of their expressions with precancerous and paracancerous esophageal lesions and esophageal carcinoma.METHODS:In situ hybridization and immunohistochemistry were used respectively to detect the expression of mRNA and proteins of Egr-1, c-fos and cyclin D1 in 70 cases of esophageal squamous cell carcinoma and their corresponding para-cancerous mucosa and upper cut edge mucosa.RESULTS:In situ hybridization and immunohistochemistry showed positive staining of all three mRNAs in the cytoplasm and those of the proteins in nuclei. Overexpression of Egr-1, c-los and cyclin D1 mRNAs and their proteins was found in dysplasia and squamous carcinomas. The expression level of Egr-1 and c-los was high, and cyclin D1 was low in dysplasia mucosa, whereas the expression of Egr-1 was decreased, c-fos was maintained and cyclin D1 was increased in the cancers. The expression of both c-fos and cyclinD1 was consistent between the mRNA and protein in their corresponding high expression lesions.CONCLUSION: The expression of Egr-1, c-fos and cyclin D1 varies in esophageal precancerous lesions and cancer tissues, suggesting an involvement of these genes in the development of esophageal carcinoma.
基金supported by agrant from the National Natural Science Foundation of China(No.30570546)
文摘This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellu-lar growth, apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro. The expression of TRAIL and endostatin protein in ECV304 cells was detected by ELISA after the transfection of recombinant plasmid pshuttle-Egr1-shTRAIL-shES and X-ray irradiation. Then MTT assay was used for determining the cellular proliferation, and flow cytometry (FCM) plus Annexin V and propidium iodide (PI) double-staining or PI single-staining were employed for the detection of apoptosis and cell cycle progression. The results showed that expression of TRAIL and endostatin protein exhibited a time- and dose-dependent change in ECV304 cells after pshut-tle-Egr1-shTRAIL-shES transfection in conjunction with irradiation. In the TRAIL-endostatin-based single- or double-gene-radiotherapy, the cell viability declined in a time- and dose-dependent manner, the percentage of cells at G2/M phase and apoptotic rate was increased, and the percentage of cells at G0/G1 phase was lowered as compared with those receiving radiotherapy alone. Moreover, TRAIL-endostatin-based double-gene-radiotherapy demonstrated better effects on growth inhibition, promotion of apoptosis and induction of cell cycle arrest in ECV304 cells than single-gene-radiotherapy.
文摘目的 :克隆小鼠内皮抑素 (m Endostatin)编码区 c DNA序列并构建含 Egr- 1启动子的 IFNγ和 m Endo-statin双基因表达载体。方法 :利用逆转录多聚酶链反应法 (RT- PCR) ,以小鼠肝细胞 m RNA为模板 ,扩增获得全长 m Endostatin,与 p MD1 8T载体连接作全自动测序 ,并利用基因重组技术构建含 Egr- 1启动子的 IFNγ和m Endostatin双基因表达质粒。结果 :经测序证实获得的 m Endostatin序列与文献报道完全一致 ,并构建了含 Egr-1启动子的 IFNγ和 m Endostatin双基因表达质粒 p Egr- IFNγ- m Endostatin。结论 :利用 RT- PCR法成功克隆了m Endostatin的 c DNA序列 ,构建了 p Egr- IFNγ- m Endostatin重组双基因表达质粒。
