AIM: To investigate the expression patterns of TTYH2 in the human colon cancer and colon cancer cell lines and to evaluate the inhibitory effect of small interfering RNA (siRNA) on the expression of TTYH2 in colon can...AIM: To investigate the expression patterns of TTYH2 in the human colon cancer and colon cancer cell lines and to evaluate the inhibitory effect of small interfering RNA (siRNA) on the expression of TTYH2 in colon cancer cell lines.METHODS: We investigated the expression patterns of TTYH2 in colon cancer, adjacent non-tumorous colon mucosa, and cancer cell lines (DLD-1, caco-2, and Lovo) by RT-PCR. Furthermore, a siRNA plasmid expression vector against TTYH2 was constructed and transfected into DLD-1 and Caco-2 with LipofectamineTM 2000. The down regulation of TTYH2 expression was detected by RT-PCR and the role of siRNA in inducing cell proliferation and cell aggregation was evaluated by MTT and aggregation assay.RESULTS: TTYH2 gene expression in colon cancer tissue was significantly up-regulated compared with normal colonic mucosa (1.23 ± 0.404 vs 0.655 ± 0.373, P = 0.0103). Colon cancer derived cell lines including DLD-1, Caco-2, and Lovo also expressed high levels of TTYH2. In contrast, transfection with siRNA-TTYH2 signifi cantly inhibited both proliferation and scattering of these cancer cell lines.CONCLUSION: The present work demonstrates, for the fi rst time, that the TTYH2 gene expression is signifi cantly up-regulated in colon cancer. The TTYH2 gene may play an important role in regulating both proliferating andmetastatic potentials of colorectal cancer.展开更多
The Small ubiquitin-related modifier （SUMO） conjugation to a variety of proteins regulates diverse cellular processes, including transcription, cell cycle regulation and maintenance of genome integrity. To investiga...The Small ubiquitin-related modifier （SUMO） conjugation to a variety of proteins regulates diverse cellular processes, including transcription, cell cycle regulation and maintenance of genome integrity. To investigate in vivo biological function of SUMO paralogs, we inactivated them in the early development of zebrafish. While zebrafish embryos deficient for all three SUMO paralogs, as Ubc9-deficient ones, displayed severe defects, loss of individual SUMO paralog was compatible with a normal development. SUMO-deficient embryos can be rescued by a single human or zebrafish SUMO. While key structural basic lysine residues and N-terminal unstructured stretch of SUMO are critical for in vivo rescue, the consensus Kll sumoylation site of SUMO2 is dispensable, implying that chain formation on this potential site is unessential for normal development. Inactivation of all three SUMOs triggered p53- dependent apoptosis and further inactivation of p53 restored normal zebrafish development. Interestingly, we also demonstrate that the dominant negative truncated form of p53, Δ113p53, significantly blunts SUMO depletion-induced p53 activity in vivo. Taken together, our results suggest that SUMO paralogs are indispensable, but redundant, in the early development of zebrafish.展开更多
Objective: To investigate the effect of cytokines (TNF-α,IFN-γ and IL-6) on the expression of sodium-iodide symporter(NIS)gene in breast cancer cell(MCF-7). Methods:The breast cancer cell was cultureds with negativ...Objective: To investigate the effect of cytokines (TNF-α,IFN-γ and IL-6) on the expression of sodium-iodide symporter(NIS)gene in breast cancer cell(MCF-7). Methods:The breast cancer cell was cultureds with negative control culture or culture with different concentrations of cytokines for 72 h.NIS gene mRNA in breast cancer cells cultured was determined by reverse transcriptase-polymerase chain reaction(RT-PCR). Results:Expression of sodium-iodide symporter mRNA can be found decreasing along with increasing the concentration of cytokine dose-dependently. Conclusion: Cytokine may play a role in iodide-uptake modulating in breast cancer cells by their effect on NIS gene expression.展开更多
文摘AIM: To investigate the expression patterns of TTYH2 in the human colon cancer and colon cancer cell lines and to evaluate the inhibitory effect of small interfering RNA (siRNA) on the expression of TTYH2 in colon cancer cell lines.METHODS: We investigated the expression patterns of TTYH2 in colon cancer, adjacent non-tumorous colon mucosa, and cancer cell lines (DLD-1, caco-2, and Lovo) by RT-PCR. Furthermore, a siRNA plasmid expression vector against TTYH2 was constructed and transfected into DLD-1 and Caco-2 with LipofectamineTM 2000. The down regulation of TTYH2 expression was detected by RT-PCR and the role of siRNA in inducing cell proliferation and cell aggregation was evaluated by MTT and aggregation assay.RESULTS: TTYH2 gene expression in colon cancer tissue was significantly up-regulated compared with normal colonic mucosa (1.23 ± 0.404 vs 0.655 ± 0.373, P = 0.0103). Colon cancer derived cell lines including DLD-1, Caco-2, and Lovo also expressed high levels of TTYH2. In contrast, transfection with siRNA-TTYH2 signifi cantly inhibited both proliferation and scattering of these cancer cell lines.CONCLUSION: The present work demonstrates, for the fi rst time, that the TTYH2 gene expression is signifi cantly up-regulated in colon cancer. The TTYH2 gene may play an important role in regulating both proliferating andmetastatic potentials of colorectal cancer.
基金Supplementary information is linked to the online version of the paper on the Cell Research website.Acknowledgements We thank Dr Jiang Zhu （Shanghai institute of hematology, Rui Jin hospital） and Dr Nelly Kieffer （CNRS LIA, Rui Jin hospital） for their comments. This work was supported by grants from the National High Tech Program of China （863, 2006AA02Z150）, the National Science Foundation of China （30525006）, the Science and Technology Commission of Shanghai Municipality （07XD14022, 06PJ14068）, ATIP program and BNP PARIBAS.
文摘The Small ubiquitin-related modifier （SUMO） conjugation to a variety of proteins regulates diverse cellular processes, including transcription, cell cycle regulation and maintenance of genome integrity. To investigate in vivo biological function of SUMO paralogs, we inactivated them in the early development of zebrafish. While zebrafish embryos deficient for all three SUMO paralogs, as Ubc9-deficient ones, displayed severe defects, loss of individual SUMO paralog was compatible with a normal development. SUMO-deficient embryos can be rescued by a single human or zebrafish SUMO. While key structural basic lysine residues and N-terminal unstructured stretch of SUMO are critical for in vivo rescue, the consensus Kll sumoylation site of SUMO2 is dispensable, implying that chain formation on this potential site is unessential for normal development. Inactivation of all three SUMOs triggered p53- dependent apoptosis and further inactivation of p53 restored normal zebrafish development. Interestingly, we also demonstrate that the dominant negative truncated form of p53, Δ113p53, significantly blunts SUMO depletion-induced p53 activity in vivo. Taken together, our results suggest that SUMO paralogs are indispensable, but redundant, in the early development of zebrafish.
文摘Objective: To investigate the effect of cytokines (TNF-α,IFN-γ and IL-6) on the expression of sodium-iodide symporter(NIS)gene in breast cancer cell(MCF-7). Methods:The breast cancer cell was cultureds with negative control culture or culture with different concentrations of cytokines for 72 h.NIS gene mRNA in breast cancer cells cultured was determined by reverse transcriptase-polymerase chain reaction(RT-PCR). Results:Expression of sodium-iodide symporter mRNA can be found decreasing along with increasing the concentration of cytokine dose-dependently. Conclusion: Cytokine may play a role in iodide-uptake modulating in breast cancer cells by their effect on NIS gene expression.
