Zhang G.,Chongqing Medical University |
Wen K.,Chongqing Medical University |
Zhang X.,Teaching and Research Section of Histology and Embryology |
Wang Y.,Chongqing Medical University |
And 2 more authors.
Chinese Journal of Clinical Oncology | Year: 2012
Objective: To explore the effect of Na +-K +-ATPase subunit expression on the proliferation and invasion of human colorectal cancer cell lines SW480 and SW620 and to determine the potential molecular mechanisms of ouabain anticancer therapeutics in human colorectal cancer. Methods: SW620 (Dukes C phase) and SW480 (Dukes B phase) cells were derived from the same patient and treated with or without a physiological concentration of 1 nM of ouabain. The proliferation of SW620 and SW480 cells was evaluated using an MTT reduction assay (i.e., 3-[ 4, 5]-dimethylthiahiazo [-z-yl]-3, 5-di- phenytetrazoliumromide). The invasion capacity was detected using a Transwell chamber. The Na +-K +-ATPase activity was measured using biochemistry and enzymology. The expression of the Na +-K +-ATPase α 1-, α 1-, and α 3-subunits were determined using real-time quantitative PCR and Western blot analysis. Results: The proliferation and invasion of SW620 cells significantly increased compared with those of SW480 cells (P < 0.05 to P < 0.01). The enzymatic activity of Na +-K +-ATPase remarkably decreased in SW620 cells (P < 0.001). Furthermore, in SW620 cells, the mRNA-level expression of Na +-K +-ATPase α 3-subunits was significantly downregulated compared with that of the α 1 and α 2-subunits. However, the Na +-K +-ATPase α1-subunit expression was significantly upregulated (P < 0.01). Interestingly, the Na +-K +-ATPase α 3-subunit expression in SW480 cells was upregulated (P < 0.001). Nevertheless, the Na +, K +-ATPase α2-subunit expressions in the SW480 and SW620 cells were not significantly different (P > 0.05). In addition, the protein expression of the Na + -K +-ATPase α 1, α 2-, and α 3-subunits were in line with the accompanying mRNA expression. A physiological concentration of 1 nM of ouabain, a Na +, K +-ATPase inhibitor, could significantly downregulate the protein-level expression of N +-K +-ATPase α 1-subunits in SW620 and SW480 cells within 48 h, thus inhibiting the growth and decreasing the invasion capacity of SW620 and SW480 cells. Ouabain could increase Na +-K +-ATPase activity in SW620 cells. The Na +-K +-ATPase α 2- and α 3-subunit expression in the SW620 and SW480 cells treated with ouabain for 48 h were not significantly different. Conclusion: The aberrant activity of Na +-K +-ATPase might involve tumor metastasis in human colorectal cancer, which is partly ascribed to the abnormal expression of the Na +-K +-ATPase α 1- and α 3-subunits. However, ouabain could effectively decrease the proliferation and invasion of SW480 and SW620 cells by downregulating the Na +-K +-ATPase α1-subunits.
Zhang L.,Teaching and Research Section of Histology and Embryology |
Zhang J.-K.,Teaching and Research Section of Histology and Embryology |
Guo M.,Teaching and Research Section of Histology and Embryology
Acta Anatomica Sinica | Year: 2014
Objective: To observe the expression of Bcl-2 and Bax in the development of C57/BL6 mice hippocampus. Methods: Mice hippocampus of Embryos (E) 18 day (d), E 20 d, postnatal (P) 1 d, P 3 d, P 7 d, P 14 d, P 28 d, P 2 month (M), P3 M, P6 M, P15 M, P18 M were used in this study. Immunohistochemistry and stereoscopy were used to detect the expression of Bcl-2 and Bax protein in the mice hippocampus at different ages. Results: From E18 d to P21 d, Bcl-2 and Bax immunoreactive cells in both Ammon' cornu (CA) and dentate gyrus (DG) firstly and gradually increased and then decreased (P < 0.01). The volume density of both Bcl-2 and Bax immunoreactive cells came to the top on P7 d except the Bax expression in DG which was the highest on P14 d(P < 0.01). Conclusion: The expression of Bcl-2 and Bax is high at the early stage of hippocampus development and then low at its adult stage and senectitude, which may be involved with the forming of hippocampus.