Objective:Deleted in liver cancer 1(DLC1)is a GTPase-activating protein that is reported as a suppressor in certain human cancers.However,the detailed biological function of DLC1 is still unclear in human prostate can...Objective:Deleted in liver cancer 1(DLC1)is a GTPase-activating protein that is reported as a suppressor in certain human cancers.However,the detailed biological function of DLC1 is still unclear in human prostate cancer(PCa).In the present study,we aimed to explore the function of DLC1 in PCa cells.Methods:Silencing and overexpression of DLC1 were induced in an androgen-sensitive PCa cell line(LNCaP)using RNA interference and lentiviral vector transduction.The Cell Counting Kit-8 assay was performed to determine cell proliferation.The cell cycle was examined by performing a propidium iodide staining assay.Results:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of LNCaP cells.Moreover,DLC1 expression was negatively correlated with Rho-associated protein kinase(ROCK)expression in LNCaP cells.Importantly,this study showed that the ROCK inhibitor Y27632 restored the function of DLC1 in LNCaP cells and reduced the tumorigenicity of LNCaP cells in vivo.Conclusion:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of PCa cells and negatively correlated with ROCK expression in PCa cells and tissue.展开更多
目的:探索肝癌缺失因子1(deleted in liver cancer 1,DLC-1)对耐恩杂鲁胺前列腺癌(prostate cancer,PCa)细胞增殖,侵袭和凋亡的影响以及作用机制。方法:体外以不同浓度的恩杂鲁胺药物培养人前列腺癌lncap及C4-2细胞株6个月以上,构建恩...目的:探索肝癌缺失因子1(deleted in liver cancer 1,DLC-1)对耐恩杂鲁胺前列腺癌(prostate cancer,PCa)细胞增殖,侵袭和凋亡的影响以及作用机制。方法:体外以不同浓度的恩杂鲁胺药物培养人前列腺癌lncap及C4-2细胞株6个月以上,构建恩杂鲁胺耐药细胞系lncap-R,C4-2-R。CCK-8法检测其IC50,验证恩杂鲁胺耐药株是否构建成功。利用慢病毒转染lncap-R,C4-2-R,以敲低和过表达DLC-1,采用克隆实验、Transwell实验和流式细胞术检测DLC-1对耐恩杂鲁胺的前列腺癌细胞的增殖、侵袭和凋亡的影响。结果:与对照组相比,过表达组DLC-1显著抑制了2个耐药株的增殖和侵袭进程。机制上,WB实验结果表明DLC-1表达与2个耐药株中Rho蛋白的表达呈负相关。Rho抑制剂rhosin逆转了因敲低DLC-1后lncap-R细胞凋亡减少的情况。结论:DLC-1可以下调耐药株中的Rho蛋白的表达,并抑制耐药株的增殖和侵袭。展开更多
Chronic hepatitis B is a global health problem. The clinical outcomes of chronic hepatitis B infection include asymptomatic carrier state, chronic hepatitis(CH), liver cirrhosis(LC), and hepatocellular carcinoma(HCC)....Chronic hepatitis B is a global health problem. The clinical outcomes of chronic hepatitis B infection include asymptomatic carrier state, chronic hepatitis(CH), liver cirrhosis(LC), and hepatocellular carcinoma(HCC). Because of the spontaneous error rate inherent to viral reverse transcriptase, the hepatitis B virus(HBV) genome evolves during the course of infection under the antiviral pressure of host immunity. The clinical significance of pre-S/S variants has become increasingly recognized in patients with chronic HBV infection. Pre-S/S variants are often identified in hepatitis B carriers with CH, LC, and HCC, which suggests that these naturally occurring pre-S/S variants may contribute to the development of progressive liver damage and hepatocarcinogenesis. This paper reviews the function of the pre-S/S region along with recent findings related to the role of pre-S/S variants in liver diseases. According to the mutation type, five pre-S/S variants have been identified: pre-S deletion, pre-S point mutation, pre-S1 splice variant, C-terminus S point mutation, and pre-S/S nonsense mutation. Their associations with HBV genotype and the possible pathogenesis of pre-S/S variants are discussed. Different pre-S/S variants cause liver diseases through different mechanisms. Most cause the intracellular retention of HBV envelope proteins and induction of endoplasmic reticulum stress, which results in liver diseases. Pre-S/S variants should be routinely determined in HBV carriers to help identify individuals who may be at a high risk of less favorable liver disease progression. Additional investigations are required to explore the molecular mechanisms of the pre-S/S variants involved in the pathogenesis of each stage of liver disease.展开更多
AIM To investigate the associations of different types of pre-S deletions with hepatitis B virus(HBV) genotypes.METHODS The sequences of the pre-S region, basal core promoter(BCP) mutation, and precore(PC) mutation we...AIM To investigate the associations of different types of pre-S deletions with hepatitis B virus(HBV) genotypes.METHODS The sequences of the pre-S region, basal core promoter(BCP) mutation, and precore(PC) mutation were examined through direct DNA sequencing or clonal analysis and sequencing in 273 HBV carriers, namely 55 asymptomatic carriers, 55 carriers with chronic hepatitis(CH), 55 with liver cirrhosis(LC), 53 with liver cirrhotic hepatocellular carcinoma(LC-HCC), and 55 with noncirrhotic HCC. A total of 126 HBV carriers(46.2%) harbored pre-S deletions. The DNA sequences of pre-S deletion mutants from 43 age-matched genotype B(HBV/B)-infected carriers and 43 agematched genotype C(HBV/C)-infected carriers were further examined, aligned, and compared.RESULTS No significant difference was observed in the mean age distribution(P = 0.464), male sex(P = 0.805), viral load(P = 0.635), or BCP mutation(P = 0.117) between the HBV/B and HBV/C groups. However, the rate of PC mutation was significantly higher in the HBV/B-infected carriers than in the HBV/C-infected carriers(P = 0.003). Both genotypes exhibited a high rate of deletion in the C-terminal half of the pre-S1 region and N-terminus of the pre-S2 region(86.0% and 79.1% in the HBV/B group; 69.8% and 72.1% in the HBV/C group, respectively). Epitope mapping showed that deletion in several epitope sites was frequent i n both genotypes, particularly p S1-BT and p S2-B2. Conversely, the rate of p S2-B1 deletion was significantly higher in the HBV/B group(72.1% vs 37.2%, P = 0.002), and the rate of pS 2-T deletion was significantly higher in the HBV/C group(48.8% vs 25.6%, P = 0.044). Functional mapping showed that the rate of deletion in three functional sites(the nucleocapsid binding site, start codon of M, and site for viral secretion) located in the N-terminus of the pre-S2 region was significantly higher in the HBV/B group(P < 0.05). One type of N-terminus pre-S1 deletion mutant with deletion of the start codon of the L protein was frequently observed in the HBV/C group(20.9% vs 9.3%, P = 0.228), particularly in the LC patients(42.9% vs 12.5%). Different patterns of pre-S deletions were also found between the HBV/B and HBV/C groups according to different clinical outcomes. In CH patients, deletion in the site for polymerized human serum albumin was more frequent in the HBV/B group(88.9% vs 36.4%, P = 0.028). In the LC-HCC patients, the rate of deletion in the pre-S2 region was significantly higher in the HBV/B group than in the HBV/C group(P < 0.05).CONCLUSION HBV/B- and HBV/C-infected carriers exhibit different patterns of pre-S deletion, which may be associated with the progression of liver diseases.展开更多
肝癌缺失基因-1(deleted in liver cancer-1,DLC-1)在多种肿瘤中呈现表达缺失或表达下调,这种异常表达主要与由DNA甲基转移酶(DNA methyltransferases,DNMTs)参与的启动子区异常甲基化有关。RT-PCR结果显示DLC-1在永生化鼻咽上皮细胞NP6...肝癌缺失基因-1(deleted in liver cancer-1,DLC-1)在多种肿瘤中呈现表达缺失或表达下调,这种异常表达主要与由DNA甲基转移酶(DNA methyltransferases,DNMTs)参与的启动子区异常甲基化有关。RT-PCR结果显示DLC-1在永生化鼻咽上皮细胞NP69和干扰DNMTs的5-8F细胞中的表达水平较未干扰的鼻咽癌细胞明显升高。甲基化特异性PCR(methylation-specific PCR,MSPCR)结果则表明DLC-1启动子区在表达下调或缺失的鼻咽癌细胞中均存在异常高甲基化,而干扰DNMTs后5-8F细胞中DLC-1启动子区甲基化状态被逆转,其中特异性干扰DNMT1后效果略为显著,提示DNA甲基转移酶活性对于鼻咽癌中DLC-1启动子区甲基化水平具有重要的调控作用,而DNMT1的调控作用更为突出。展开更多
目的探讨乳腺癌缺失基因1(deleted in breast cancer 1,DBC1)对肝癌细胞增殖和凋亡的影响。方法构建pcDNA3.1-DBC1的正义表达载体,转染肝癌细胞SMMC-7721,MTT法检测DBC1对肝癌细胞SMMC-7721生长增殖的影响,采用流式细胞术(FCM)检测DBC1...目的探讨乳腺癌缺失基因1(deleted in breast cancer 1,DBC1)对肝癌细胞增殖和凋亡的影响。方法构建pcDNA3.1-DBC1的正义表达载体,转染肝癌细胞SMMC-7721,MTT法检测DBC1对肝癌细胞SMMC-7721生长增殖的影响,采用流式细胞术(FCM)检测DBC1对肝癌细胞SMMC-7721的细胞周期和细胞凋亡的影响。结果与转染空载体的SMMC-7721细胞相比,pcDNA3.1-DBC1正义表达载体转染上调DBC1的表达,可显著抑制肝癌细胞(SMMC-7721)增殖,升高G1期细胞比例(51.0%vs 64.9%,P=0.002),降低S期细胞比例(29.7%vs 14.0%,P<0.001),诱导细胞凋亡(8.8%vs 24.6%,P<0.001)。结论 DBC1通过抑制肿瘤细胞增殖,阻滞细胞周期进展,促进细胞凋亡,抑制肝癌的发生和发展。展开更多
肝癌缺失基因(deleted in liver cancer genes,DLCs)家族存在于人体多种组织中,其表达产物为RhoGTP酶活化蛋白。该基因家族在多种肿瘤中低表达或不表达,主要通过负性调控Rho蛋白(RhoA,Cdc42)和黏着斑蛋白及两者下游效应因子的活性,抑制...肝癌缺失基因(deleted in liver cancer genes,DLCs)家族存在于人体多种组织中,其表达产物为RhoGTP酶活化蛋白。该基因家族在多种肿瘤中低表达或不表达,主要通过负性调控Rho蛋白(RhoA,Cdc42)和黏着斑蛋白及两者下游效应因子的活性,抑制细胞骨架形成、黏着斑重组,在肿瘤细胞生长、增殖、侵袭和转移过程中发挥重要作用。雌、孕激素和胰岛素可通过转录后修饰途径调控DLCs蛋白活性,在此类激素相关肿瘤的发生发展中发挥作用。展开更多
基金This study was supported by the Key Scientific Research Project of Shanghai Municipal Commission of Health and Family Planning(No.201640014)the project of Natural Science Foundation of Jiangxi(No.20171BAB205019)the Special Diseases Program of Pudong New Area Health System(No.PWZzb2017-06).
文摘Objective:Deleted in liver cancer 1(DLC1)is a GTPase-activating protein that is reported as a suppressor in certain human cancers.However,the detailed biological function of DLC1 is still unclear in human prostate cancer(PCa).In the present study,we aimed to explore the function of DLC1 in PCa cells.Methods:Silencing and overexpression of DLC1 were induced in an androgen-sensitive PCa cell line(LNCaP)using RNA interference and lentiviral vector transduction.The Cell Counting Kit-8 assay was performed to determine cell proliferation.The cell cycle was examined by performing a propidium iodide staining assay.Results:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of LNCaP cells.Moreover,DLC1 expression was negatively correlated with Rho-associated protein kinase(ROCK)expression in LNCaP cells.Importantly,this study showed that the ROCK inhibitor Y27632 restored the function of DLC1 in LNCaP cells and reduced the tumorigenicity of LNCaP cells in vivo.Conclusion:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of PCa cells and negatively correlated with ROCK expression in PCa cells and tissue.
文摘目的:探索肝癌缺失因子1(deleted in liver cancer 1,DLC-1)对耐恩杂鲁胺前列腺癌(prostate cancer,PCa)细胞增殖,侵袭和凋亡的影响以及作用机制。方法:体外以不同浓度的恩杂鲁胺药物培养人前列腺癌lncap及C4-2细胞株6个月以上,构建恩杂鲁胺耐药细胞系lncap-R,C4-2-R。CCK-8法检测其IC50,验证恩杂鲁胺耐药株是否构建成功。利用慢病毒转染lncap-R,C4-2-R,以敲低和过表达DLC-1,采用克隆实验、Transwell实验和流式细胞术检测DLC-1对耐恩杂鲁胺的前列腺癌细胞的增殖、侵袭和凋亡的影响。结果:与对照组相比,过表达组DLC-1显著抑制了2个耐药株的增殖和侵袭进程。机制上,WB实验结果表明DLC-1表达与2个耐药株中Rho蛋白的表达呈负相关。Rho抑制剂rhosin逆转了因敲低DLC-1后lncap-R细胞凋亡减少的情况。结论:DLC-1可以下调耐药株中的Rho蛋白的表达,并抑制耐药株的增殖和侵袭。
基金Supported by the grant from the National Science Council(NSC 96-2320-B-030-004-MY3),Executive Yuan,Taiwan
文摘Chronic hepatitis B is a global health problem. The clinical outcomes of chronic hepatitis B infection include asymptomatic carrier state, chronic hepatitis(CH), liver cirrhosis(LC), and hepatocellular carcinoma(HCC). Because of the spontaneous error rate inherent to viral reverse transcriptase, the hepatitis B virus(HBV) genome evolves during the course of infection under the antiviral pressure of host immunity. The clinical significance of pre-S/S variants has become increasingly recognized in patients with chronic HBV infection. Pre-S/S variants are often identified in hepatitis B carriers with CH, LC, and HCC, which suggests that these naturally occurring pre-S/S variants may contribute to the development of progressive liver damage and hepatocarcinogenesis. This paper reviews the function of the pre-S/S region along with recent findings related to the role of pre-S/S variants in liver diseases. According to the mutation type, five pre-S/S variants have been identified: pre-S deletion, pre-S point mutation, pre-S1 splice variant, C-terminus S point mutation, and pre-S/S nonsense mutation. Their associations with HBV genotype and the possible pathogenesis of pre-S/S variants are discussed. Different pre-S/S variants cause liver diseases through different mechanisms. Most cause the intracellular retention of HBV envelope proteins and induction of endoplasmic reticulum stress, which results in liver diseases. Pre-S/S variants should be routinely determined in HBV carriers to help identify individuals who may be at a high risk of less favorable liver disease progression. Additional investigations are required to explore the molecular mechanisms of the pre-S/S variants involved in the pathogenesis of each stage of liver disease.
基金Supported by the National Science Council,Taiwan,No.NSC 96-2320-B-030-004-MY3
文摘AIM To investigate the associations of different types of pre-S deletions with hepatitis B virus(HBV) genotypes.METHODS The sequences of the pre-S region, basal core promoter(BCP) mutation, and precore(PC) mutation were examined through direct DNA sequencing or clonal analysis and sequencing in 273 HBV carriers, namely 55 asymptomatic carriers, 55 carriers with chronic hepatitis(CH), 55 with liver cirrhosis(LC), 53 with liver cirrhotic hepatocellular carcinoma(LC-HCC), and 55 with noncirrhotic HCC. A total of 126 HBV carriers(46.2%) harbored pre-S deletions. The DNA sequences of pre-S deletion mutants from 43 age-matched genotype B(HBV/B)-infected carriers and 43 agematched genotype C(HBV/C)-infected carriers were further examined, aligned, and compared.RESULTS No significant difference was observed in the mean age distribution(P = 0.464), male sex(P = 0.805), viral load(P = 0.635), or BCP mutation(P = 0.117) between the HBV/B and HBV/C groups. However, the rate of PC mutation was significantly higher in the HBV/B-infected carriers than in the HBV/C-infected carriers(P = 0.003). Both genotypes exhibited a high rate of deletion in the C-terminal half of the pre-S1 region and N-terminus of the pre-S2 region(86.0% and 79.1% in the HBV/B group; 69.8% and 72.1% in the HBV/C group, respectively). Epitope mapping showed that deletion in several epitope sites was frequent i n both genotypes, particularly p S1-BT and p S2-B2. Conversely, the rate of p S2-B1 deletion was significantly higher in the HBV/B group(72.1% vs 37.2%, P = 0.002), and the rate of pS 2-T deletion was significantly higher in the HBV/C group(48.8% vs 25.6%, P = 0.044). Functional mapping showed that the rate of deletion in three functional sites(the nucleocapsid binding site, start codon of M, and site for viral secretion) located in the N-terminus of the pre-S2 region was significantly higher in the HBV/B group(P < 0.05). One type of N-terminus pre-S1 deletion mutant with deletion of the start codon of the L protein was frequently observed in the HBV/C group(20.9% vs 9.3%, P = 0.228), particularly in the LC patients(42.9% vs 12.5%). Different patterns of pre-S deletions were also found between the HBV/B and HBV/C groups according to different clinical outcomes. In CH patients, deletion in the site for polymerized human serum albumin was more frequent in the HBV/B group(88.9% vs 36.4%, P = 0.028). In the LC-HCC patients, the rate of deletion in the pre-S2 region was significantly higher in the HBV/B group than in the HBV/C group(P < 0.05).CONCLUSION HBV/B- and HBV/C-infected carriers exhibit different patterns of pre-S deletion, which may be associated with the progression of liver diseases.
文摘目的探讨乳腺癌缺失基因1(deleted in breast cancer 1,DBC1)对肝癌细胞增殖和凋亡的影响。方法构建pcDNA3.1-DBC1的正义表达载体,转染肝癌细胞SMMC-7721,MTT法检测DBC1对肝癌细胞SMMC-7721生长增殖的影响,采用流式细胞术(FCM)检测DBC1对肝癌细胞SMMC-7721的细胞周期和细胞凋亡的影响。结果与转染空载体的SMMC-7721细胞相比,pcDNA3.1-DBC1正义表达载体转染上调DBC1的表达,可显著抑制肝癌细胞(SMMC-7721)增殖,升高G1期细胞比例(51.0%vs 64.9%,P=0.002),降低S期细胞比例(29.7%vs 14.0%,P<0.001),诱导细胞凋亡(8.8%vs 24.6%,P<0.001)。结论 DBC1通过抑制肿瘤细胞增殖,阻滞细胞周期进展,促进细胞凋亡,抑制肝癌的发生和发展。
文摘肝癌缺失基因(deleted in liver cancer genes,DLCs)家族存在于人体多种组织中,其表达产物为RhoGTP酶活化蛋白。该基因家族在多种肿瘤中低表达或不表达,主要通过负性调控Rho蛋白(RhoA,Cdc42)和黏着斑蛋白及两者下游效应因子的活性,抑制细胞骨架形成、黏着斑重组,在肿瘤细胞生长、增殖、侵袭和转移过程中发挥重要作用。雌、孕激素和胰岛素可通过转录后修饰途径调控DLCs蛋白活性,在此类激素相关肿瘤的发生发展中发挥作用。
