AIM: To investigate the alteration of the annexin I subcellular localization in esophageal squarnous cell carcinoma (ESCC)and the correlation between the translocation and the tumorigenesis of ESCC.METHODS: The protei...AIM: To investigate the alteration of the annexin I subcellular localization in esophageal squarnous cell carcinoma (ESCC)and the correlation between the translocation and the tumorigenesis of ESCC.METHODS: The protein localization of annexin I was detected in both human ESCC tissues and cell line via the indirect immunofluorescence strategy.RESULTS: In the normal esophageal epithelia the annex in I was mainly located on the plasma membrane and formed a consecutive typical trammels net. Annexin I protein also expressed dispersively in cytoplasm and the nuclei without specific localization on the nuclear membrane. In esophageal cancer annexin I decreased very sharply with scattered disappearance on the cellular membrane, however it translocated and highly expressed on the nuclear membrane,which was never found in normal esophageal epithelia. In cultured esophageal cancer cell line annexin I protein was also focused on the nuclear membrane, which was consistent with the result from esophageal cancer tissues.CONCLUSION: This observation suggests that the translocation of annexin I protein in ESCC may correlate with the tumorigenesis of the esophageal cancer.展开更多
AIM: To investigate biogenesis and intracellular Iocalizations of clusterin to elucidate the potential molecular mechanisms implicated in tumorigenesis of esophageal mucosa.METHODS: Semi-quantitative RT-PCR for multi-...AIM: To investigate biogenesis and intracellular Iocalizations of clusterin to elucidate the potential molecular mechanisms implicated in tumorigenesis of esophageal mucosa.METHODS: Semi-quantitative RT-PCR for multi-region alteration analysis, Western blot for different transcriptional forms and immunohistochemical staining for intracellular Iocalizations of clusterin were carried out in both tissues and cell lines of ESCC.RESULTS: The N-terminal deletions of the clusterin gene and the appearance of a 50-53 ku nuclear clusterin, an uncleaved, nonglycosylated, and disulfide-linked isoform,were the major alterations in cancer cells of esophagus.Naturally the 40 ku clusterin was located in the connective tissue of the lamina propria of epithelial mucosa and right under the basal membrane of epithelia, but it was disappeared in stromal mucosa of esophagus and the pre-matured clusterin was found positive in cancerous epithelia.CONCLUSION: The N-terminal deletion of clusterin may be essential for its alterations of biogenesis in ESCC.展开更多
AIM: To investigate gene expression pattern of human γsynuclein gene in human esophageal squamous cell carcinoma (ESCC) by using semi-quantitive reverse transcription polymerase chain reaction (RT-PCR), and to study ...AIM: To investigate gene expression pattern of human γsynuclein gene in human esophageal squamous cell carcinoma (ESCC) by using semi-quantitive reverse transcription polymerase chain reaction (RT-PCR), and to study the role of γ-synudein in the development of human ESCC.METHODS: Semi-quantitive RT-PCR of 27 pairs of specimens of human ESCC tissues and corresponding normal tissues was used to investigate the expression pattern of γsynuclein in ESCC. 9706/γ-syn cells in which γ-synuclein was overexpressed were obtained through cloning γ-synuclein gene by PCR and transfecting it into ESCC 9706 cells, then selecting with G-418 for 14 days. The biological effects of γsynuclein were measured and compared between 9706/γ-syn and 9706/vec cells by cell growth curve and soft agar assay.RESULTS: RT-PCR showed that γ-synuclein gene was expressed in all the 27 cases of normal epithelial tissues,while downregulation of γ-synudein was observed in 16 out of the 27 cases (59.3%) of ESCC. There were also 6 cases of ESCC tissues with a high expression level of γ-synuclein mRNA. In functional analysis we found that over-expression of γ-synuclein in ESCC 9706 cells could inhibit the growth rate and transformation ability of ESCC 9706 cells.CONCLUSION: The low expression level of γ-synuclein in human ESCC and the biological effects of γ-synuclein overexpression on ESCC 9706 cells suggest that γ-synuclein may play a role as a negative regulator in the development of human ESCC.展开更多
基金the Major State Basic Research Development Program of China,No.G1998051205the National Hi-Tech R & D Program of China,No.2001AA227091+1 种基金the National Natural Science Foundation of China,No.39990570(Major Program)and No.30171049(General Program)the National Science Fund for Distinguished Young Scholars(No.30225045)
文摘AIM: To investigate the alteration of the annexin I subcellular localization in esophageal squarnous cell carcinoma (ESCC)and the correlation between the translocation and the tumorigenesis of ESCC.METHODS: The protein localization of annexin I was detected in both human ESCC tissues and cell line via the indirect immunofluorescence strategy.RESULTS: In the normal esophageal epithelia the annex in I was mainly located on the plasma membrane and formed a consecutive typical trammels net. Annexin I protein also expressed dispersively in cytoplasm and the nuclei without specific localization on the nuclear membrane. In esophageal cancer annexin I decreased very sharply with scattered disappearance on the cellular membrane, however it translocated and highly expressed on the nuclear membrane,which was never found in normal esophageal epithelia. In cultured esophageal cancer cell line annexin I protein was also focused on the nuclear membrane, which was consistent with the result from esophageal cancer tissues.CONCLUSION: This observation suggests that the translocation of annexin I protein in ESCC may correlate with the tumorigenesis of the esophageal cancer.
基金Supported by National Natural Science Foundation,No.30225045,No.39990570,No.30171049 and No.30370713,and National High Tech and Major State Basic R & D Program of China,No.G 1998051205and No.2001AA227091
文摘AIM: To investigate biogenesis and intracellular Iocalizations of clusterin to elucidate the potential molecular mechanisms implicated in tumorigenesis of esophageal mucosa.METHODS: Semi-quantitative RT-PCR for multi-region alteration analysis, Western blot for different transcriptional forms and immunohistochemical staining for intracellular Iocalizations of clusterin were carried out in both tissues and cell lines of ESCC.RESULTS: The N-terminal deletions of the clusterin gene and the appearance of a 50-53 ku nuclear clusterin, an uncleaved, nonglycosylated, and disulfide-linked isoform,were the major alterations in cancer cells of esophagus.Naturally the 40 ku clusterin was located in the connective tissue of the lamina propria of epithelial mucosa and right under the basal membrane of epithelia, but it was disappeared in stromal mucosa of esophagus and the pre-matured clusterin was found positive in cancerous epithelia.CONCLUSION: The N-terminal deletion of clusterin may be essential for its alterations of biogenesis in ESCC.
基金the National Natural Science Foundation of China, No.39925020State Key Basic Research Program,No.G1998051204
文摘AIM: To investigate gene expression pattern of human γsynuclein gene in human esophageal squamous cell carcinoma (ESCC) by using semi-quantitive reverse transcription polymerase chain reaction (RT-PCR), and to study the role of γ-synudein in the development of human ESCC.METHODS: Semi-quantitive RT-PCR of 27 pairs of specimens of human ESCC tissues and corresponding normal tissues was used to investigate the expression pattern of γsynuclein in ESCC. 9706/γ-syn cells in which γ-synuclein was overexpressed were obtained through cloning γ-synuclein gene by PCR and transfecting it into ESCC 9706 cells, then selecting with G-418 for 14 days. The biological effects of γsynuclein were measured and compared between 9706/γ-syn and 9706/vec cells by cell growth curve and soft agar assay.RESULTS: RT-PCR showed that γ-synuclein gene was expressed in all the 27 cases of normal epithelial tissues,while downregulation of γ-synudein was observed in 16 out of the 27 cases (59.3%) of ESCC. There were also 6 cases of ESCC tissues with a high expression level of γ-synuclein mRNA. In functional analysis we found that over-expression of γ-synuclein in ESCC 9706 cells could inhibit the growth rate and transformation ability of ESCC 9706 cells.CONCLUSION: The low expression level of γ-synuclein in human ESCC and the biological effects of γ-synuclein overexpression on ESCC 9706 cells suggest that γ-synuclein may play a role as a negative regulator in the development of human ESCC.
