ABSTRACT
Objective@#To observe the effect of Yishen Tonglong Decoction (YTD) on the epithelial-mesenchymal transition (EMT) and Ras/ERK signaling pathway in human PCa DU-145 cells and explore its action mechanism.@*METHODS@#We treated human PCa DU-145 cells with normal plasma (the blank control) or plasma containing 5% (low-dose), 10% (medium-dose) and 15% (high-dose) YTD. After intervention, we examined the proliferation of the DU-145 cells in different groups with CCK-8 and their apoptosis by Annexin V/PI double staining. We detected the cell cycle by PI assay, the invasion and migration of the cells using the Transwell chamber and scratch test, and the expressions of the proteins and genes related to the EMT and Ras/ERK signaling pathways in the cells by Western blot and RT-PCR.@*RESULTS@#Compared with the blank control group, high-, medium- and low-dose YTD significantly inhibited the proliferation of the PCa DU-145 cells, decreased their adherence and growth (P < 0.05, P < 0.01), promoted their apoptosis (P < 0.01), regulated their cell cycles (P < 0.05, P < 0.01), and reduced their in vitro invasion and migration abilities (P < 0.05), all in a dose-dependent manner. The results of Western blot and RT-PCR revealed down-regulated protein and mRNA expressions of N-cadherin, zinc finger transcription factor (Snail), Ras, p-ERK1/2 and ERK1/2, but up-regulated protein and mRNA expressions of E-cadherin in the PCa DU-145 cells treated with YTD (P < 0.05, P < 0.01).@*CONCLUSIONS@#Yishen Tonglong Decoction can effectively inhibit the proliferation, promote the apoptosis, regulate the cell cycle and suppress the invasion and migration abilities and EMT process of human PCa DU-145 cells. The mechanism of Yishen Tonglong Decoction acting on PCa may be associated with its inhibitory effect on the EMT process and expression of the Ras/ERK signaling pathway in PCa cells./.
Subject(s)
Humans , Male , Drugs, Chinese Herbal , Epithelial-Mesenchymal Transition , Prostatic Neoplasms , Signal TransductionABSTRACT
@#[Abstract] Objective: To study the effect of microRNA-141(miR-141) expression regulation on cell proliferation, cell cycle, apoptosis, invasion and migration of human prostate cancer cell line DU145 and its mechanism. Methods: MiR-141 mimics (miR-141 up group) and miR-141 inhibitors (miR-141 down group) were transfected into human prostate cancer DU145 cells by using liposome lipofectamine 2000, and the un-transfection group (Control group) and non-sense miRNA sequence transfection group (NC group) were set. The expression of miRNA-141 in DU145 cells in each group before and after transfection was detected by qPCR. MTT assay was used to detect the proliferation viability and sensitivity to cisplatin (DDP) in DU145 cells of each group. Cell cycle and apoptosis rate of DU145 cells under DDP treatment were detected by Flow cytometry; the changes in cell invasion and migration ability were detected by Transwell method. The protein expressions of VEGF and EGFR in DU145 cells of each group were detected by Western blotting. Results: Compared with the Control and NC group, the level of miRNA-141 expression in the miR-141-down group decreased to (0.18±0.08), the cell proliferation viability decreased significantly while its sensitivity to DDP increased significantly, the cell cycle was blocked in the G0+G1 phase, and the apoptosis rate significantly increased to (46.67±5.86)% while cell invasion rate and migration rate significantly decreased to (44.34±8.32)%, (57.73±6.19)%, and the relative expression levels of VEGF and EGFR decreased to (0.47±0.06), (0.36±0.06), (P<0.05 or P<0.01). But in the miR-141-up group, the level of miRNA-141 expression increased to (4.23±0.53), the cell proliferation viability significantly increased while its sensitivity to DDP decreased significantly, and the cell cycle was promoted into S and G2 phase, the apoptosis rate significantly decreased to (18.77±4.24)% while cell invasion and migration rate significantly increased to (89.94±6.34)%, (94.44±5.84)%, and the relative expression levels of VEGF and EGFR were up to (0.89±0.07), (0.73±0.06),(P<0.05 or P<0.01). Conclusion: miR-141 can act as a growth promoting factor in prostate cancer DU145 cells. miR-141 down-regula‐tion can significantly inhibit the proliferation viability, cell cycle, migration and invasion of DU145 cells, and promote cell apoptosis and DDP-sensitivity, and the mechanism of which may be related with inhibition of VEGF and EGFR protein expressions.
ABSTRACT
Objective:To investigate the anti-tumor effect of Hyperoside.Methods: Human γδT cells were amplified by isopentenyl pyrophosphate from peripheral blood cells.The proliferation capacity of γδT cells was measured with CCK-8 assay after treated with different concentrations of Hyperoside.Cytotoxicity of γδT cells was detected with LDH assay , and the expression of granzyme,perforin CD107a and IFN-γonγδT cells were measured by flow cytometry before and after treatment.Results: Hyperoside could significantly stimulate the proliferation of γδT cells at the concentration of 3.13-12.5 μg/ml.Cytotoxicity and expression of granzyme,perforin and IFN-γofγδT cells were increased after treatment.Conclusion:Hyperoside could enhance cytotoxicity of humanγδT cells through up-regulation of granzyme ,perforin CD107 a and IFN-γexpression.