ABSTRACT
Background: Colon cancer is a major cause of morbidity and mortality in the world. Juglone is a natural compound which has been isolated from Juglans mandshurica Maxim, and it has various pharmacological effects such as antiviral, antibacterial, and anticancer. In our study, we aimed to investigate the effect of juglone on CCL-228-SW 480 colon carcinoma cell line in monolayer and spheroid culture medium. Materials and Methods: The CCL-228-SW 480 cell lines were cultured in both monolayer and spheroid cultures. Cells were treated with juglone at 24, 48, and 72 h of incubation. ID50 inhibition was determined on the dose for juglone and after it was found 20 μM was applied to the cells to examine the effect of juglone on CCL-228-SW 480 colon carcinoma cell line. After Juglone was applied the BrdU marking index, Transferase dUTP Nick ends Labeling (TUNEL) assay, active caspase-3 assay, apoptosis-inducing factor (AIF) assay were determined by immunohistochemistry in both the monolayer and spheroid cultures. Results: The control group had a healthy pattern of S-phase fraction, and many of the CCL-228-SW 480 cells nuclei were observed to be positive for BrdU. Terminal Deoxynucleotidyl TUNEL-positive cells, active Caspase-3, and AIF were detected after treatment with juglone in both the monolayer and spheroid cultures. Conclusions: The dead cell count was higher in the CCL-228-SW 480 cell lines with juglone applied than in the controls. Juglone significantly inhibits the proliferation and induces the apoptosis of CCL-228-SW 480 cells in vitro
ABSTRACT
Background and purpose:As a new candidate tumor suppressor gene, p33ING1b has many biological functions. This study was done to investigate its role in the regulation of the proliferation of human colon cancer cell line SW480. Methods:The pcDNA3.1(+)/p33ING1b/SW480 cells were identified by Western blot and S-P immunohistochemical method. In order to elucidate the effect of expression of exogenous p33ING1b gene on the colorectal cancer cell SW480, the proliferation rates were analyzed by growth curves and colony formation assay in soft agar for cells including SW480, pcDNA3.1(+)/p33ING1b/SW480 and pcDNA3.1(+)/SW480. At same time, the apoptotic rate of cells and the cell cycle analysis were also tested by flow cytometry.Furthermore,using western blot analysis,we detected the expression level of the protein p53, p21WAF1,Bax and Bcl-2 in those three group cells, which initially indicate the molecule mechanism of inducing apoptosis by gene p33ING1b. Results:The cell growth rates of SW480 cells transfected with pcDNA3.1 (+)/p33ING1b were slower than those transfected with pcDNA3.1(+) or untransfected.The colony formation efficiency of pcDNA3.1(+)/p33ING1b/SW480 were decreased and the apoptotic rates were increased compared with pcDNA3.1(+)/SW480 and SW480(P0.05). Conclusion:After overexpression of exogenous p33ING1b protein in SW480 cell, there were inhibition of the proliferation rate and induction of apoptosis, the molecular mechanism might be associated with up- regulated expression of Bax and down-regulated expression of Bcl-2 by p33ING1b gene.
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Purpose:To investigate the effect of short hairpin RNA (shRNA) targeting CD44v3 on adhesion and invasion behavior of human colon cancer cell line SW480 induced by hyaluronan in vitro. Methods:RNA interference plasmid including U6 promoter and expressing shRNA of CD44v3 was designed, constructed, and transfected into SW480 cell line. CD44v3 expression was examined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Adhesive and invasive ability were examined by plate adhesion model and Boyden chamber model.Results:After plasmid transfection,CD44v3 expression in SW480 cell and the number of SW480 cell induced by hyaluronan adhesiving on plate or permeating septum of Boyden chamber decreased significantly (P
ABSTRACT
Aim To investigate the arrest effect of diallyl disulfide(DADS) in the cell cycle of human nasopharyngeal carcinoma SW480 cell line and its molecular mechanism.Mothdes The growth inhibition effect of DADS of different concentrations on SW480 cell line was measured by MTT assay and cell counting.Phase distribution of cell cycle was analyzed by flow cytometry.Expression of Ubiquitin and FKBP was determined by Western blot.Results The MTT assay showed that DADS inhibited growth of SW480 cells significantly in a dose-dependent manner.Adding 25,35,50 and 70 mg?L-1 DADS for 48 hours,SW480 cell growth was suppressed by 18.67%,33.02%,49.12% and 66.86%,respectively.There were significant differences between the treated and controlled cases(P