Summary: This study examined the effect of artesunate (Art) on the proliferation, DNA replication, cell cycles and apoptosis of vascular smooth muscle cells (VSMCs). Primary cultures of VSMCs were established from aor...Summary: This study examined the effect of artesunate (Art) on the proliferation, DNA replication, cell cycles and apoptosis of vascular smooth muscle cells (VSMCs). Primary cultures of VSMCs were established from aortas of mice and artesunate of different concentrations was added into the medium. The number of VSMCs was counted and the curve of cell growth was recorded. The activity of VSMCs was assessed by using MTT method and inhibitory rate was calculated. DNA replication was evaluated by [3H]-TdR method and apoptosis by DNA laddering and HE staining. Flowmetry was used for simultaneous analysis of cell apoptosis and cell cycles. Compared with the control group, VSMCs proliferation in Art interfering groups were inhibited and [3H]-TdR incorprating rate were decreased as well as cell apoptosis was induced. The progress of cell cycle was blocked in G 0/G 1 by Art in a dose-dependent manner. It is concluded that Art inhibits VSMCs proliferation by disturbing DNA replication, inducing cell apoptosis and blocking cell cycle in G 0/G 1 phase.展开更多
Background:This study aimed to select compounds with unique inhibitory effects on muscle-invasive bladder cancer(MIBC)from coumarone derivatives with similar parent nuclear structures and to reveal their tumor-suppres...Background:This study aimed to select compounds with unique inhibitory effects on muscle-invasive bladder cancer(MIBC)from coumarone derivatives with similar parent nuclear structures and to reveal their tumor-suppressive effects using various approaches.Methods:Bladder cancer cell lines SW780 and T24,as well as human normal bladder epithelial cell line SV-HUC-1 were selected as the study model,and these urinary system cells were co-incubated with various concentrations of(S,E)-4-(4-methylbenzylidene)-3-phenylchroman-3-ol,(S,E)-4-(4-isocyanobenzylidene)-3-phenylchroman-3-ol,(S,E)-4-(4-fluorobenzylidene)-3-phenylchroman-3-ol(FPO),and(S,E)-3-phenyl-4-(4-(trifluoromethoxy)benzylidene)chroman-3-ol.Cell activity was detected using cell counting kit-8.FPO showed the strongest inhibitory effect on MIBC cells;therefore,it was selected for further experiments.We monitored the FPO-induced T24 cell morphological changes with an inverted microscope.The FPO-inhibited migration of T24 cells was examined using a cell scratch assay.We detected the clonogenic ability of T24 cells through a clone formation test and evaluated their proliferative ability using a 5-ethynyl-2’-deoxyuridine fluorescence staining kit.The inhibitory effect of FPO against the cell cycle was monitored using flow cytometry,and its suppressive effect on the DNA replication ability of T24 cells was detected using double fluorescence staining(Ki67 and phalloidin).Results:Among the four candidate coumarone derivatives,FPO showed the most significant inhibitory effect on MIBC cells and was less toxic to normal urothelial cells.FPO inhibited T24 cell growth in time and dose-dependent manners(the half-inhibitory concentration is 8μM).FPO significantly repressed the proliferation,migration,and clonogenic ability of bladder cancer T24 cells.Cell mobility was significantly inhibited by FPO:30μM FPO almost completely repressed migration occurred at after 24 h treatment.Moreover,FPO significantly suppressed the clonogenicity of bladder cancer cells in a dose-dependent manner.Mechanistically,FPO targeted the cell cycle,arresting the S and G2 phases on bladder cancer T24 cells.Conclusion:We discovered a novel anticancer chemical,FPO,and proposed a potential mechanism,through which it suppresses MIBC T24 cells by repressing the cell cycle in the S and G2 phases.This study contributes to the development of novel anticancer drugs for MIBC.展开更多
Objective: To research the inhibitory effect on SGC-7901 cells of α-pinene, and the related mechanism of α-pinene. Methods: Used the MTT method to detect inhibition rate and western blotting to detect the influence ...Objective: To research the inhibitory effect on SGC-7901 cells of α-pinene, and the related mechanism of α-pinene. Methods: Used the MTT method to detect inhibition rate and western blotting to detect the influence on expression of ATM, Phos-S1981ATM, H2AX, γH2AX, CHK2 and p-CHK2, p53 and phos-p53 cell cycle related protein in SGC-7901 cells. Results: The research found α-pinene could inhibit the proliferation of SGC-7901 cells observably in vitro, and the inhibition rate assumes the dependence on concentration;and western blotting results showed that, α-pinene could activate phospho-ATM, increase the amount of γH2AX (p p < 0.05);increase the expression of p-CHK2, p53 and phos-p53 (p p p < 0.05);But there is no significant effect on expression of CHK2 (p > 0.05). Conclusions: α-pinene could inhibit the proliferation of SGC-7901 cells, and by inducing ATM (Ataxia Telangiectasia-Mutated) kinase signal pathway in DNA damage response, activating cell cycle checkpoint, making the cell cycle arrest then exerts its anti-tumor effects.展开更多
The proliferation of vascular smooth muscle cells(VSMCs) plays a major role in the pathogenesis of many cardiovascular diseases.Geminin regulates DNA replication and cell cycle progression and plays a key role in the ...The proliferation of vascular smooth muscle cells(VSMCs) plays a major role in the pathogenesis of many cardiovascular diseases.Geminin regulates DNA replication and cell cycle progression and plays a key role in the proliferation of cancer cells.We therefore hypothesized that geminin regulates the proliferation of VSMCs.The present study demonstrates that the level of geminin expression was low in quiescent VSMCs(approximately 90% and 10% of cells in the G1 and in S/G2/M phases of the cell cycle,respectively),increased as more cells entered in S/G2/M,and then decreased as cells exited S/G2/M.Further,angiotensin II and norepinephrine stimulated expression of geminin in VSMCs.However,the DNA content,nuclear morphology,percentage of cells at different stages of the cell cycle,and rate of proliferation of VSMCs from which geminin was either depleted or overexpressed were all similar.These findings indicate geminin functions differently in VSMCs than it does in cancer cell lines and that it may provide a target for treating cancers without affecting normal cells.展开更多
Background: DNA replication and sister chromatid cohesion 1 (DSCC1) (also called DCC1) is a component of an alternative replication factor C complex that loads proliferating cell nuclear antigen onto DNA during S phas...Background: DNA replication and sister chromatid cohesion 1 (DSCC1) (also called DCC1) is a component of an alternative replication factor C complex that loads proliferating cell nuclear antigen onto DNA during S phase of the cell cycle. It is located at 8q24 and frequently amplified in hepatocellular carcinoma (HCC). However, the role of DSCC1 in the carcinogenesis and progress of HCC has not been fully investigated. Here, we aimed to assert the importance of DSCC1 in the HCC. Methods: In this study, copy number variation data and RNA sequencing data were used to calculate the DNA copy number and mRNA expression of DSCC1 in HCC. Quantitative polymerase chain reaction, Western blotting, and immunohistochemistry analysis were used to determine the mRNA and protein level of DSCC1 in HCC. The Kaplan–Meier analysis and univariate and multivariate Cox regression analysis were used to assess the association of DSCC1 with the overall survival (OS) of HCC patients. Moreover, lentiviral shRNA was used to knockdown DSCC1, and then, colony?forming assay, cell cycle assay, and cell proliferation assay were performed to evaluate the impact of DSCC1 silencing on HCC cell lines. Results: We found that DSCC1 was amplified and highly expressed in HCC tumor tissues than in nontumor tissues. We then found that the overexpression of both mRNA and protein of DSCC1 was linked to the bad prognosis of HCC patients. Astonishingly, the protein level of DSCC1 was an independent prognostic factor for OS (hazard ratio, 1.79; 95% confidence interval, 1.17–2.74; P = 0.007). Furthermore, the clonogenic capacity of DSCC1?amplified HCC cell lines (MHCC?97H, MHCC?97L, and Hep3B) was significantly inhibited by transduction of a lentiviral shRNA that targets DSCC1. We also showed that knockdown of DSCC1 induced G0–G1 cell cycle arrest (increased from 60% to more than 80%) and greatly inhibited the proliferation of HCC cell lines. Conclusion: These results suggest that DSCC1 is a putative HCC driver gene that promotes proliferation and is associated with poor prognosis in HCC.展开更多
目的了解供肝复流后细胞增殖周期的变化,明确药物预处理和缺血预处理对其的影响.方法建立大鼠三袖套法原位肝移植模型,观察移植肝细胞复流前后不同时间增殖细胞核抗原的表达情况;对供肝分别作药物预处理和缺血预处理,比较正常肝、预处...目的了解供肝复流后细胞增殖周期的变化,明确药物预处理和缺血预处理对其的影响.方法建立大鼠三袖套法原位肝移植模型,观察移植肝细胞复流前后不同时间增殖细胞核抗原的表达情况;对供肝分别作药物预处理和缺血预处理,比较正常肝、预处理供肝和未预处理供肝复流2 h 后的增殖细胞核抗原、肝酶谱及肝细胞凋亡状况.结果移植肝细胞复流2 h 后增殖细胞核抗原表达显著增高,12 h 后恢复正常水平;正常肝、药物预处理供肝、缺血预处理供肝、未预处理供肝复流2 h 后的增殖细胞核抗原表达和肝酶谱依次升高,PCNA(%):2.1±0.1,4.3±0.2,7.0±0.4,9.4±0.4,AST(nKat·L^(-1)):3.5±1.6,28.1±11.9,53.8±12.3,75.2±24.1,ALT(nKat·L^(-1)):2.8±2.7,46.1±17.6,61.7±22.9,90.5±30.1,LDH(nKat·L^(-1)):49.6±7.6,61.4±15.3,95.4±23.2,148.3±30.1.结论供肝复流后肝细胞 G_1期缩短至2 h 以内,整个细胞周期时间缩短至20 h 左右.正常肝、药物预处理供肝、缺血预处理供肝、未预处理供肝肝损害程度依次加重,整个细胞周期时间依次缩短.展开更多
文摘Summary: This study examined the effect of artesunate (Art) on the proliferation, DNA replication, cell cycles and apoptosis of vascular smooth muscle cells (VSMCs). Primary cultures of VSMCs were established from aortas of mice and artesunate of different concentrations was added into the medium. The number of VSMCs was counted and the curve of cell growth was recorded. The activity of VSMCs was assessed by using MTT method and inhibitory rate was calculated. DNA replication was evaluated by [3H]-TdR method and apoptosis by DNA laddering and HE staining. Flowmetry was used for simultaneous analysis of cell apoptosis and cell cycles. Compared with the control group, VSMCs proliferation in Art interfering groups were inhibited and [3H]-TdR incorprating rate were decreased as well as cell apoptosis was induced. The progress of cell cycle was blocked in G 0/G 1 by Art in a dose-dependent manner. It is concluded that Art inhibits VSMCs proliferation by disturbing DNA replication, inducing cell apoptosis and blocking cell cycle in G 0/G 1 phase.
基金supported by National Nature Science Foundation of China(82172978)Taishan Scholars Program of Shandong Province(Grant No.tsqn201909147)+1 种基金the Key Project at Central Government Level:the ability establishment of sustainable use for valuable Chinese medicine resources(2060302)the Student Innovation Training Program in Jining Medical University(cx2021116).
文摘Background:This study aimed to select compounds with unique inhibitory effects on muscle-invasive bladder cancer(MIBC)from coumarone derivatives with similar parent nuclear structures and to reveal their tumor-suppressive effects using various approaches.Methods:Bladder cancer cell lines SW780 and T24,as well as human normal bladder epithelial cell line SV-HUC-1 were selected as the study model,and these urinary system cells were co-incubated with various concentrations of(S,E)-4-(4-methylbenzylidene)-3-phenylchroman-3-ol,(S,E)-4-(4-isocyanobenzylidene)-3-phenylchroman-3-ol,(S,E)-4-(4-fluorobenzylidene)-3-phenylchroman-3-ol(FPO),and(S,E)-3-phenyl-4-(4-(trifluoromethoxy)benzylidene)chroman-3-ol.Cell activity was detected using cell counting kit-8.FPO showed the strongest inhibitory effect on MIBC cells;therefore,it was selected for further experiments.We monitored the FPO-induced T24 cell morphological changes with an inverted microscope.The FPO-inhibited migration of T24 cells was examined using a cell scratch assay.We detected the clonogenic ability of T24 cells through a clone formation test and evaluated their proliferative ability using a 5-ethynyl-2’-deoxyuridine fluorescence staining kit.The inhibitory effect of FPO against the cell cycle was monitored using flow cytometry,and its suppressive effect on the DNA replication ability of T24 cells was detected using double fluorescence staining(Ki67 and phalloidin).Results:Among the four candidate coumarone derivatives,FPO showed the most significant inhibitory effect on MIBC cells and was less toxic to normal urothelial cells.FPO inhibited T24 cell growth in time and dose-dependent manners(the half-inhibitory concentration is 8μM).FPO significantly repressed the proliferation,migration,and clonogenic ability of bladder cancer T24 cells.Cell mobility was significantly inhibited by FPO:30μM FPO almost completely repressed migration occurred at after 24 h treatment.Moreover,FPO significantly suppressed the clonogenicity of bladder cancer cells in a dose-dependent manner.Mechanistically,FPO targeted the cell cycle,arresting the S and G2 phases on bladder cancer T24 cells.Conclusion:We discovered a novel anticancer chemical,FPO,and proposed a potential mechanism,through which it suppresses MIBC T24 cells by repressing the cell cycle in the S and G2 phases.This study contributes to the development of novel anticancer drugs for MIBC.
文摘Objective: To research the inhibitory effect on SGC-7901 cells of α-pinene, and the related mechanism of α-pinene. Methods: Used the MTT method to detect inhibition rate and western blotting to detect the influence on expression of ATM, Phos-S1981ATM, H2AX, γH2AX, CHK2 and p-CHK2, p53 and phos-p53 cell cycle related protein in SGC-7901 cells. Results: The research found α-pinene could inhibit the proliferation of SGC-7901 cells observably in vitro, and the inhibition rate assumes the dependence on concentration;and western blotting results showed that, α-pinene could activate phospho-ATM, increase the amount of γH2AX (p p < 0.05);increase the expression of p-CHK2, p53 and phos-p53 (p p p < 0.05);But there is no significant effect on expression of CHK2 (p > 0.05). Conclusions: α-pinene could inhibit the proliferation of SGC-7901 cells, and by inducing ATM (Ataxia Telangiectasia-Mutated) kinase signal pathway in DNA damage response, activating cell cycle checkpoint, making the cell cycle arrest then exerts its anti-tumor effects.
基金supported by Beijing Municipal Natural Science Foundation (5102040)
文摘The proliferation of vascular smooth muscle cells(VSMCs) plays a major role in the pathogenesis of many cardiovascular diseases.Geminin regulates DNA replication and cell cycle progression and plays a key role in the proliferation of cancer cells.We therefore hypothesized that geminin regulates the proliferation of VSMCs.The present study demonstrates that the level of geminin expression was low in quiescent VSMCs(approximately 90% and 10% of cells in the G1 and in S/G2/M phases of the cell cycle,respectively),increased as more cells entered in S/G2/M,and then decreased as cells exited S/G2/M.Further,angiotensin II and norepinephrine stimulated expression of geminin in VSMCs.However,the DNA content,nuclear morphology,percentage of cells at different stages of the cell cycle,and rate of proliferation of VSMCs from which geminin was either depleted or overexpressed were all similar.These findings indicate geminin functions differently in VSMCs than it does in cancer cell lines and that it may provide a target for treating cancers without affecting normal cells.
基金grants from the National Natural Science Foundation of China (No.81201569and No.81541151)the Beijing Natural Science Foundation (No.7132186)the National Key Sci-Tech Special Project of China (No.2018ZX10302207and No.2017ZX10203202).
文摘Background: DNA replication and sister chromatid cohesion 1 (DSCC1) (also called DCC1) is a component of an alternative replication factor C complex that loads proliferating cell nuclear antigen onto DNA during S phase of the cell cycle. It is located at 8q24 and frequently amplified in hepatocellular carcinoma (HCC). However, the role of DSCC1 in the carcinogenesis and progress of HCC has not been fully investigated. Here, we aimed to assert the importance of DSCC1 in the HCC. Methods: In this study, copy number variation data and RNA sequencing data were used to calculate the DNA copy number and mRNA expression of DSCC1 in HCC. Quantitative polymerase chain reaction, Western blotting, and immunohistochemistry analysis were used to determine the mRNA and protein level of DSCC1 in HCC. The Kaplan–Meier analysis and univariate and multivariate Cox regression analysis were used to assess the association of DSCC1 with the overall survival (OS) of HCC patients. Moreover, lentiviral shRNA was used to knockdown DSCC1, and then, colony?forming assay, cell cycle assay, and cell proliferation assay were performed to evaluate the impact of DSCC1 silencing on HCC cell lines. Results: We found that DSCC1 was amplified and highly expressed in HCC tumor tissues than in nontumor tissues. We then found that the overexpression of both mRNA and protein of DSCC1 was linked to the bad prognosis of HCC patients. Astonishingly, the protein level of DSCC1 was an independent prognostic factor for OS (hazard ratio, 1.79; 95% confidence interval, 1.17–2.74; P = 0.007). Furthermore, the clonogenic capacity of DSCC1?amplified HCC cell lines (MHCC?97H, MHCC?97L, and Hep3B) was significantly inhibited by transduction of a lentiviral shRNA that targets DSCC1. We also showed that knockdown of DSCC1 induced G0–G1 cell cycle arrest (increased from 60% to more than 80%) and greatly inhibited the proliferation of HCC cell lines. Conclusion: These results suggest that DSCC1 is a putative HCC driver gene that promotes proliferation and is associated with poor prognosis in HCC.
文摘目的了解供肝复流后细胞增殖周期的变化,明确药物预处理和缺血预处理对其的影响.方法建立大鼠三袖套法原位肝移植模型,观察移植肝细胞复流前后不同时间增殖细胞核抗原的表达情况;对供肝分别作药物预处理和缺血预处理,比较正常肝、预处理供肝和未预处理供肝复流2 h 后的增殖细胞核抗原、肝酶谱及肝细胞凋亡状况.结果移植肝细胞复流2 h 后增殖细胞核抗原表达显著增高,12 h 后恢复正常水平;正常肝、药物预处理供肝、缺血预处理供肝、未预处理供肝复流2 h 后的增殖细胞核抗原表达和肝酶谱依次升高,PCNA(%):2.1±0.1,4.3±0.2,7.0±0.4,9.4±0.4,AST(nKat·L^(-1)):3.5±1.6,28.1±11.9,53.8±12.3,75.2±24.1,ALT(nKat·L^(-1)):2.8±2.7,46.1±17.6,61.7±22.9,90.5±30.1,LDH(nKat·L^(-1)):49.6±7.6,61.4±15.3,95.4±23.2,148.3±30.1.结论供肝复流后肝细胞 G_1期缩短至2 h 以内,整个细胞周期时间缩短至20 h 左右.正常肝、药物预处理供肝、缺血预处理供肝、未预处理供肝肝损害程度依次加重,整个细胞周期时间依次缩短.
