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Biomedical subjects

Da-xing Xie

Publications and source records attributed to Da-xing Xie.

3 recordsLinked to original sources

[PTEN induces anoikis through its phosphatase activity in hepatocellular carcinoma cells].

OBJECTIVE: To investigate the effect and mechanisms of tumor suppressor gene PTEN on the induction of anoikis of hepatocellular carcinoma SMMC-7721 cells. METHODS: SMMC-7721 cells were transfected with GFP plasmids containing wild-type PTEN or phosphatase inactivating mutant PTEN (C124A-PTEN) in vitro; The PTEN expression and the phosphorylation levels of focal adhesion kinase (FAK) and protein kinase B (PKB/Akt) were detected by Western blotting; Flow cytometry assay and laser scanning confocal microscopy were used to analyze apoptosis in adherent and non-adherent cells. RESULTS: Compared with the control, PTEN expression in the cells transfected with wild-type PTEN increased to 248%, while the phosphorylation level of FAK and Akt decreased 65.2% and 89.1%, respectively; and the anoikis percentage increased from 9.5% to 31.3%. In the cells transfected with C124A-PTEN, neither the phosphorylation of FAK and Akt nor the anoikis percentage had obviously changed, although the PTEN expression enhanced dramatically in comparison with the control. CONCLUSION: Through its phosphatase activity, tumor suppressor gene PTEN can suppress the phosphorylation of FAK and Akt, and induce anoikis in hepatocellular carcinoma cells.

Anoikis↗

Peplomycin induces G1-phase specific apoptosis in liver carcinoma cell line Bel-7402 involving G2-phase arrest.

AIM: To investigate the mechanism of peplomycin (PEP)-induced apoptosis in liver carcinoma cell line (Bel-7402). METHODS: Growth inhibition by PEP was analyzed using 3- 4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptotic cells were detected using Hoechest 33258 staining, and confirmed by flow cytometric analysis and DNA fragmentation analysis. The expression of cyclin A and B1 were determined by flow cytometry and Western blot. Annexin V assay was measured by flow cytometric analysis. RESULTS: PEP induced apoptosis and then inhibited cell proliferation in liver carcinoma cell line Bel-7402. Cells treated with PEP 50 mumol/L for 15 h were arrested in G2-phase with dramatical expression of cyclin A and a little change in cyclin B1. Almost all the apoptosis occurred in cells undergoing the G1-phase after treatment for 24 h. CONCLUSION: Peplomycin induced G1-phase specific apoptosis in Bel-7402 involving G2-phase arrest.

Antibiotics, Antineoplastic↗

[Quantitative analysis of cell cyclin E expression threshold].

BACKGROUND & OBJECTIVES: It is important to analyze the threshold of cyclins when we research the mechanism of cell cycle progressing. However, there was no effective way to caculate quantitatively. This study was designed to analyze cyclin E expression threshold quantitatively. METHODS: MOLT-4 cells were detected at different photomultiplier tube (PMT) voltage by cyclin E/DNA multiparameter flow cytometry. Using this method, MOLT-4 cells were detected at the same voltage after being treated with caffeine and cycloheximide (CHX), and then MOLT-4 cells and JURKAT cells were detected at the same voltage. Threshold of cyclin E was counted by using formula B2/A x C (A, B, C indicates the minimum, threshold, and maximum of cyclin E fluorescence intensity respectively). RESULTS: Cyclin E threshold of MOLT-4 cells calculated by formula B2/A x C was invariable at different voltage. It decreased when cells treaded with caffeine and unchanged when treated with cycloheximide. At the same time, cyclin E threshold of JURKAT cells calculated by formula was much lower than that of Molt-4 cells. CONCLUSIONS: Formula B2/A x C can be used to analyze cyclin E expression threshold quantitatively.

Caffeine↗