RESUMO
ANXA2, a member of the annexin family, is overexpressed and plays important roles in tumor development. However, the significance of ANXA2 expression in gastric carcinoma has not been clarified.To elucidate its roles in growth of gastric cancer, ANXA2 expression in SGC-7901 cells was inhibited with a designated siRNA, then cell proliferation, cell cycling, apoptosis and motility were determined by MTT assay, flow cytometry, Hoechst 33342 staining and wound healing assay, respectively. To further assess the behavior of ANXA2 deleted SGC- 7901 cells, changes of microstructures were observed under fluorescence microscopy, laser scanning confocal microscopy and electron microscopy. We found that inhibition of ANXA2 expression caused cell proliferation to decrease significantly with G1 arrest, motility to be reduced with changes in pseudopodia/filopodia structure and F-actin and ß-tubulin expression, and apoptosis to be enhanced albeit without significance. At the same time, ANXA2 deletion resulted in fewer pseudopodia/filopodia, non-stained areas were increased, contact inhibition among cells reappeared, and expression of F-actin and ß-tubulin was decreased, with induction of polymerized disassembled forms. Taken together, these data suggest that ANXA2 overexpression is important to maintain the malignancy of cancer cells, and this member of the annexin family has potential to be considered as a target for the gene therapy of gastric carcinoma.
Assuntos
Anexina A2/metabolismo , Apoptose , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Actinas/genética , Actinas/metabolismo , Anexina A2/antagonistas & inibidores , Anexina A2/genética , Western Blotting , Ciclo Celular , Humanos , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Gástricas/genética , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo , Células Tumorais Cultivadas , CicatrizaçãoRESUMO
Numerous research results have suggested that the events occurred in a selected cell target and the fates of the cells are spatiotemporally regulated. It has been paying much attention to study the cell events with spatiotemporal proposal designation and analysis. We have been tracking and thinking the new scientific area for many years. Spatiotemporal cell biology and its schematic frame, triple W (when, where, which), are systemically introduced in this study. The triple W under pathological conditions is also discussed.
Assuntos
Biologia Celular/tendências , Fenômenos Fisiológicos Celulares , Animais , Expressão Gênica , Humanos , Resolução de Problemas , Fatores de TempoRESUMO
A beta-1,3-glucanase was detected, using laminarin as substrate, in the culture broth of Chaetomium sp. Major activity was associated with a 70 kDa protein band visualized on a polyacrylamide gel. beta-1,3-Glucanase was purified by a one-step, native gel purification procedure. Optimal activity was observed at pH 6.0 and 30 degrees C (over 30 min). It could degrade cell walls of plant pathogens including Rhizoctonia solani, Gibberella zeae, Fusarium sp., Colletotrichum gloeosporioides and Phoma sp. The N-terminal amino acid residues of the purified beta-1,3-glucanase are PYQLQTP, which do not exhibit homology to other fungal beta-1,3-glucanases suggesting it may be a novel enzyme.
