A Synthetic Biscoumarin Suppresses Lung Cancer Cell Proliferation and Induces Cell Apoptosis by Increasing Expression of RIP1
2
Citation
38
Reference
10
Related Paper
Citation Trend
Abstract:
Coumarin has a variety of biological activities and widely exists in plants. Biscoumarin, derived from coumarin, their synthetic methods and bioactivities of biscoumarins is the hotspot of the current research. In this study, we evaluated for the first time the anticancer of a synthetic biscoumarin (3,3'-(4-chlorophenyl)methylene)bis(4-hydroxy-2H-chromen-2-one, C3) on lung cancer cells and explored the related mechanism. C3 was simply prepared by 4-hydroxycoumarin and 4-chlorobenzaldehyde under ethanol. The structure of C3 was elucidated by various spectroscopic analyses. The antiproliferation effect of C3 was evaluated by the cell counting kit-8 assay. Cell cycle and apoptosis analysis were detected by flow cytometry. The expression of correlated proteins was determined using Western blotting. The result showed that C3 displayed a strong cytostatic effect on Lewis lung cancer (LLC) cells. C3 inhibited the proliferation of LLC cells, and induced G2/M phase cell cycle arrest. In addition, C3 possessed a significant reduction on cell apoptosis by increasing of RIP1 expression. Our data showed that C3 suppresses lung cancer cell proliferation and induces cell apoptosis, which is possibly involved with the RIP1.Background: Ligustrazine, active ingredients extracted from the natural herb Ligusticum Chuanxiong Hort, has promising anti-tumor properties on tumor cell lines. However, the potential anti-tumor activity of ligustrazine on colorectal cancer (CRC) cells and the molecular mechanisms have not been elucidated. In this study, we explored the critical functions of ligustrazine on SW480 and CT26 cells at cellular levels. Methods: CCK-8 assay was performed to analyze the cell viability. Flow cytometry analysis was applied to study cell apoptosis and cell cycle. The expressions of cell apoptosis and cell cycle-associated proteins were conducted by western blot and qRT-PCR analysis. Results: Ligustrazine showed significant inhibitory effects on the proliferation of SW480 and CT26 cells. Ligustrazine induced cell apoptosis was associated with the up-regulation of pro-apoptotic protein and the down-regulation of anti-apoptotic protein in an activated mitochondrial-dependent pathway. And it indicated that ligustrazine induced cell cycle arrest by changing the cell cycle distribution, which leads to cell cycle arrest at the G0/G1 phase. Besides, the ligustrazine-induced cell apoptosis and cell cycle arrest were markedly reversed by pifithrin-α (p53 inhibitor), which suggested that ligustrazine-induced cell apoptosis was achieved by regulating p53-dependent mitochondrial pathway and cell cycle arrest at the G0/G1 phase. Conclusions: These findings demonstrated that ligustrazine could induce SW480 and CT26 cells apoptosis via a p53-dependent mitochondrial pathway and cell cycle arrest at the G0/G1 phase. Ligustrazine may serve as a potential anti-cancer agent for CRC.
G1 phase
Viability assay
Cite
Citations (19)
Objective to study the effect of SudanⅠ,Ⅲ,Ⅳ on the growth of SGC-7901.The change of DNA content is determined with Flow Cytometry,and the cell cycle is analyzed.Results show that the average content of DNA decreases in G_1 phase and increases in S and G_2 phase.The index of PI and SPF increase.It is indicated that SudanⅠ,Ⅲ,Ⅳ accelerate the cell cycle from G_1 phase to S and G_2 phase.It is also shown that the effects on SGC-7901 by Sudan Ⅲ in the dose of 50 μg·mL~(-1) and by SudanⅠ,Ⅳ in the dose of 10μg·mL~(-1) is very effective(P0.01).It is concluded that SudanⅠ,Ⅲ,Ⅳ affect SGC-7901 cell cycle to advance its differentiation and proliferation.
Cytometry
Proliferation index
Cite
Citations (0)
The levels of constitutive and inducible forms of heat shock protein 70 (hsp73 and hsp72, respectively) through the cell cycle were measured in CHO cells by flow cytometry and Western blotting at various times after heating. Cells were labeled with antibody C92 (hsp72) or N27 (hsp73) and propidium iodide prior to analysis by flow cytometry. Cells were heated for 15 min at 45°C, then analyzed from 3 to 36 h later. There was about a tenfold increase in hsp72 in early S phase cells beginning within 6 h after heating and these cells gradually cycled though S phase so by 36 h most of them had divided. When CHO cells were exposed to 10 μM sodium vanadate, an inhibitor of tyrosine phosphatase, for 24 h prior to heating, the induction of hsp72 in early S phase cells was almost completely inhibited. Heated cells did not express hsp73 in a cell-cycle-dependent manner. Hsp73 increased uniformly in all cells by 10 h after heating and sodium vanadate did not affect the expression. Quantitative comparisons of the relative levels of hsp72 and hsp73 measured by flow cytometry and Western blotting were in excellent agreement. Control and heated cells were labeled with Hoechst 33342 and sorted from G1, S, and G2/M phases and processed by Western blotting to verify the cell cycle dependent increase in hsp72 as measured by flow cytometry. Again there was excellent agreement between the Western blotting and flow cytometry results. © 1996 Wiley-Liss, Inc.
Propidium iodide
Cytometry
Cite
Citations (0)
The levels of constitutive and inducible forms of heat shock protein 70 (hsp73 and hsp72, respectively) through the cell cycle were measured in CHO cells by flow cytometry and Western blotting at various times after heating. Cells were labeled with antibody C92 (hsp72) or N27 (hsp73) and propidium iodide prior to analysis by flow cytometry. Cells were heated for 15 min at 45°C, then analyzed from 3 to 36 h later. There was about a tenfold increase in hsp72 in early S phase cells beginning within 6 h after heating and these cells gradually cycled though S phase so by 36 h most of them had divided. When CHO cells were exposed to 10 μM sodium vanadate, an inhibitor of tyrosine phosphatase, for 24 h prior to heating, the induction of hsp72 in early S phase cells was almost completely inhibited. Heated cells did not express hsp73 in a cell-cycle-dependent manner. Hsp73 increased uniformly in all cells by 10 h after heating and sodium vanadate did not affect the expression. Quantitative comparisons of the relative levels of hsp72 and hsp73 measured by flow cytometry and Western blotting were in excellent agreement. Control and heated cells were labeled with Hoechst 33342 and sorted from G1, S, and G2/M phases and processed by Western blotting to verify the cell cycle dependent increase in hsp72 as measured by flow cytometry. Again there was excellent agreement between the Western blotting and flow cytometry results. © 1996 Wiley-Liss, Inc.
Propidium iodide
Cytometry
Cite
Citations (27)
Abstract In this unit, two protocols are described for analyzing cell cycle status using flow cytometry. The first is based on the simultaneous analysis of proliferation‐specific marker (Ki‐67) and cellular DNA content, which discriminate resting/quiescent cell populations (G0 cell) and quantify cell cycle distribution (G1, S, or G2/M), respectively. The second is based on differential staining of DNA and RNA through co‐staining of Hoechst 33342 and Pyronin Y, which is also useful to identify G0 cells from G1 cells. Along with these methods for analyzing cell cycle status, two additional methods for cell proliferation assays with recent updates of newly developed fluorophores, which allow multiplex analysis of cell cycle status, cell proliferation, and a gene of interest using flow cytometry, are outlined. © 2015 by John Wiley & Sons, Inc.
Multiplex
Cytometry
Cell counting
Proliferation Marker
Cite
Citations (249)
G1 phase
Viability assay
Cite
Citations (1)
Plant-derived polyphenols are being tested as chemopreventive agents; some polyphenols arrest the cell cycle at G1 phase, whereas others inhibit cell cycle proliferation at G2/M phase. Therefore, polyphenols have been proposed to inhibit cell cycle progression at different phases via distinct mechanisms. Indeed, our previous studies showed that small structural differences in polyphenols cause large differences in their biological activities; however, the details of the structural properties causing G1 cell cycle arrest remain unknown. In this study, we prepared 27 polyphenols, including eight different scaffolds, to gain insight into the structural conditions that arrest the cell cycle at G1 phase in a quantitative structure–activity relationship study. We used cell cycle profiles to determine the biophores responsible for G1 cell cycle arrest and believe that the biophores identified in this study will help design polyphenols that cause G1 cell cycle arrest.
G1 phase
Cite
Citations (32)
Cytometry
Cite
Citations (2)
Cytometry
Cite
Citations (18)
Cell cycle arrest after different types of DNA damage can occur in either G1 phase or G2 phase of the cell cycle, involving the distinct mechanisms of p53/p21 Cip1/Waf1 induction, and phosphorylation of Cdc2, respectively. Treatment of asynchronously growing Swiss3T3 cells with the chemotherapeutic drug adriamycin induced a predominantly G2 cell cycle arrest. Here we investigate why Swiss3T3 cells were arrested in G2 phase and not in G1 phase after adriamycin‐induced damage. We show that adriamycin was capable of inducing a G1 cell cycle arrest, both during the G0‐G1 transition and during the G1 phase of the normal cell cycle. In G0 cells, adriamycin induced a prolonged cell cycle arrest. However, adriamycin caused only a transient cell cycle delay when added to cells at later time points during G0‐G1 transition or at the G1 phase of normal cell cycle. The G1 arrest correlated with the induction of p53 and p21 Cip1/Waf1 , and the exit from the arrest correlated with the decline of their expression. In contrast to the G1 arrest, adriamycin‐induced G2 arrest was relatively tight and correlated with the Thr‐14/Tyr‐15 phosphorylation of cyclin B‐Cdc2 complexes. The relative stringency of the G1 versus G2 cell cycle arrest may explain the predominance of G2 arrest after adriamycin treatment in mammalian cells.
G1 phase
Restriction point
Cyclin B1
Cite
Citations (52)