Background Considerable evidence suggests that phosphatase of regenerating liver-3 （PRL-3） plays multiple roles in cancer metastasis; however, the molecular mechanisms remain largely unknown. The aim of this study w...Background Considerable evidence suggests that phosphatase of regenerating liver-3 （PRL-3） plays multiple roles in cancer metastasis; however, the molecular mechanisms remain largely unknown. The aim of this study was to identify proteins associated with PRL-3-promoted colon cancer metastasis, by comparative proteomic analysis. Methods Proteomes of human colon cancer LoVo cells transfected with PRL-3 gene （LoVo-PRL-3） or empty vector PAcGFP-C3 （LoVo-control） were compared using 2D gel electrophoresis. Proteins that varied significantly in concentration were selected and identified using mass spectrometry. Expression of translationally controlled tumor protein （TCTP） mRNA and protein in LoVo-PRL-3 and LoVo-control cells was detected by real-time PCR and Western blotting. Small interfering RNA （siRNA） targeting TCTP was used for silencing TCTP expression in LoVo-PRL-3 cells. Functional significance of TCTP in PRL-3-promoted colon cancer cell proliferation, migration and invasion was investigated by Cell Counting Kit-8 assay and transwell chamber. Results Seventeen proteins displaying significant and reproducible differences between LoVo-PRL-3 and LoVo-control cells were identified. Ten proteins were upregulated and seven were downregulated in LoVo-PRL-3 cells when compared with LoVo-control cells. Eight identified proteins are associated with distinct steps of tumor metastasis： ubiquitin-like protein ISG15, interleukin-18, TCTP, serpin B5, annexin A3, macrophage-capping protein, ATP-dependent RNA helicase DDX3X, and cathepsin D. Real-time PCR and Western blotting results showed that both TCTP mRNA and protein were significantly increased in LoVo-PRL-3 cells compared to LoVo-control cells. Transfection with TCTP siRNA significantly reduced the expression of both mRNA and protein levels of TCTP in LoVo-PRL-3 cells. Knockdown of TCTP by siRNA inhibited PRL-3-promoted proliferation, migration and invasion of LoVo-PRL-3 cells. Conclusion Our results imply that TCTP might be a mediator of PRL-3-promoted proliferation, migration and invasion of human colon cancer cells.展开更多
AIM:To investigate the prognostic significance of phosphatase regenerating liver 3(PRL-3)protein expression in gastric cancer.METHODS:PRL-3 expression in paraffin-embedded tumor specimens from 293 patients with gastri...AIM:To investigate the prognostic significance of phosphatase regenerating liver 3(PRL-3)protein expression in gastric cancer.METHODS:PRL-3 expression in paraffin-embedded tumor specimens from 293 patients with gastric cancer was studied retrospectively by immunohistochemistry.Monoclonal antibody specifically against PRL-3,3B6,was obtained with hybridoma technique.RESULTS:Positive PRL-3 expression was detected in 43.3%(127 of 293)of gastric cancer cases.High expression of PRL-3 was positively correlated with tumor size,depth of invasion,vascular/lymphatic invasion,lymph node metastasis,high TNM stage and tumor recurrence.Patients with positive PRL-3 expression had a significantly lower 5-year survival rate than those with negative expression(28.3%vs 51.9%,P<0.0001).Patients who received curative surgery,and with positive PRL-3 expression had a significant shorter overall survival and disease-free disadvantage over patients with negative expression(hazard ratio of 16.7 and 16.6,respectively;P<0.0001 for both).Multivariate analysis revealed that PRL-3 expression was an independent prognostic indicator for overall and disease-free survival of gastric cancer patients,particularly for survival in TNM stageⅢpatients.CONCLUSION:PRL-3 expression is a new independent prognostic indicator to predict the potential of recurrence and survival in patients with gastric cancer at the time of tumor resection.展开更多
目的:研究miR495、miR551a干扰质粒对SGC7901胃癌细胞中促肝细胞再生磷酸酶-3(phosphatase of regenerating liver-3,PRL-3)表达的影响,探讨基因干扰在胃癌治疗中的价值.方法:分别体外培养稳定转染后的SGC7901细胞系及未经处理的SGC790...目的:研究miR495、miR551a干扰质粒对SGC7901胃癌细胞中促肝细胞再生磷酸酶-3(phosphatase of regenerating liver-3,PRL-3)表达的影响,探讨基因干扰在胃癌治疗中的价值.方法:分别体外培养稳定转染后的SGC7901细胞系及未经处理的SGC7901细胞系,取对数生长期瘤细胞,0.5mL(1×107/mL)细胞悬液接种于Balb/ca(nu/nu)裸鼠腹腔内,在SPF条件下饲养1mo后处死,观察裸鼠成瘤率、瘤体生长情况及腹膜转移等情况,并行组织病理学检查.取各组部分移植瘤行荧光定量PCR检测,对比各组miR495、miR551a及PRL-3mRNA的相对表达水平.结果:各组裸鼠成瘤率均100%,其中两实验组裸鼠一般情况及生存期均好于对照组.取各组移植瘤行荧光定量PCR检测显示,两实验组裸鼠miRNA表达量显著高于对照组,PRL-3mRNA表达量低于对照组.转染质粒组胃癌迁移能力明显减弱.结论:转染靶向干扰PRL-3表达的miR495、mi551a真核质粒可以明显抑制胃癌细胞体内转移侵袭能力.展开更多
胃癌是世界范围内常见肿瘤之一,因胃癌死亡的患者数占据肿瘤相关死因的第3位.肝再生磷酸酶-3(phosphatase of regenerating liver 3,PRL-3)作为一个新近发现的蛋白酪氨酸磷酸酶,近年来研究发现其在胃癌组织中高表达,在胃癌淋巴转移、腹...胃癌是世界范围内常见肿瘤之一,因胃癌死亡的患者数占据肿瘤相关死因的第3位.肝再生磷酸酶-3(phosphatase of regenerating liver 3,PRL-3)作为一个新近发现的蛋白酪氨酸磷酸酶,近年来研究发现其在胃癌组织中高表达,在胃癌淋巴转移、腹膜转移等发挥重要作用,与胃癌患者预后负相关.其后,越来越多的研究关注其在胃癌发生、发展中的调控机制,以期阐明PRL-3在胃癌发生、发展中的具体调节通路及影响因素.尽管随着研究的不断深入,对于PRL-3在胃癌中的作用机制得到一部分阐释,但对于P R L-3在促进胃癌淋巴转移、腹膜转移等恶性进展及复发的机制仍然不是很清楚,本文即对近年来PRL-3的相关研究进展作一综述.展开更多
目的观察慢病毒介导的sh RNA沉默肝再生磷酸酶-3(PRL-3)基因对结肠癌SW480细胞增殖、侵袭、凋亡的影响。方法实验分组为空白对照组、阴性对照组、转染组。将携带PRL-3 sh RNA的慢病毒载体转染结肠癌SW480细胞,建立稳定沉默PRL-3的细胞株...目的观察慢病毒介导的sh RNA沉默肝再生磷酸酶-3(PRL-3)基因对结肠癌SW480细胞增殖、侵袭、凋亡的影响。方法实验分组为空白对照组、阴性对照组、转染组。将携带PRL-3 sh RNA的慢病毒载体转染结肠癌SW480细胞,建立稳定沉默PRL-3的细胞株,real-time PCR检测转染后PRL-3 m RNA的相对表达水平。采用MTT法、平板克隆形成实验检测转染后细胞增殖能力;采用Transwell侵袭实验、侵袭小室法检测转染后细胞迁移及侵袭能力;采用流式细胞术检测转染后细胞凋亡率变化。结果稳定沉默PRL-3的细胞株构建成功,转染组PRL-3m RNA的相对表达水平低于空白对照组、阴性对照组(P<0.05),空白对照组、阴性对照组比较差异无统计学意义。PRL-3 sh RNA转染SW480细胞72 h后,转染组与空白对照组、阴性对照组比较,细胞增殖能力受到抑制,转染120 h时最明显(P<0.05)。转染组克隆形成能力较空白对照组、阴性对照组下降(P<0.05)。转染组与空白对照组、阴性对照组比较,细胞迁移、侵袭能力下降,凋亡率增加(P<0.05)。结论结肠癌SW480细胞转染PRL-3 sh RNA可减少PRL-3的表达,有效抑制SW480细胞增殖,促进其凋亡,PRL-3可能成为治疗结肠癌的靶基因。展开更多
目的:观察促肝再生磷酸酶-3(phosphatase of regeneration liver-3,PRL-3)抑制剂原钒酸钠(sodium orthovanadate,SoV)对大肠癌细胞株Colo-320迁移能力的影响.方法:应用Westernblot方法检测PRL-3在7株大肠癌细胞中的表达,筛选表达最强的...目的:观察促肝再生磷酸酶-3(phosphatase of regeneration liver-3,PRL-3)抑制剂原钒酸钠(sodium orthovanadate,SoV)对大肠癌细胞株Colo-320迁移能力的影响.方法:应用Westernblot方法检测PRL-3在7株大肠癌细胞中的表达,筛选表达最强的1株做下一步抑制实验.选择PRL-3拮抗剂SoV,通过细胞划痕实验观察其在0.5μmol/L浓度下对癌细胞运动能力的影响,高倍倒置显微镜下计算细胞迁移距离.细胞爬片原位杂交观察SoV对PRL-3mRNA的表达.结果:PRL-3表达最强的结肠癌细胞株为Colo-320.细胞划痕实验显示,SoV作用48h细胞仅迁移相当于4-7个细胞的距离,400×倍镜下精确测量20个细胞在0-48h迁移距离,计算对照组细胞迁移速度为39.12±10.11μm/h,SoV为12.84±6.78μm/h,差异非常显著(P<0.00001).SoV作用细胞48h后原位杂交结果未见PRL-3mRNA阳性表达.结论:SoV能显著抑制Colo-320细胞迁移能力,其机制可能与抑制PRL-3酶活性以及基因转录有关.展开更多
肝再生磷酸酶-3（PRL-3）属于蛋白酪氨酸磷酸酶家族，它发挥作用的方式是通过调节酪氨酸残基的磷酸化状态实现对蛋白质活性的调控。已有的研究表明，P R L-3与人多种肿瘤的发生、发展、转移及预后密切相关，可能参与调节肿瘤细胞的骨架...肝再生磷酸酶-3（PRL-3）属于蛋白酪氨酸磷酸酶家族，它发挥作用的方式是通过调节酪氨酸残基的磷酸化状态实现对蛋白质活性的调控。已有的研究表明，P R L-3与人多种肿瘤的发生、发展、转移及预后密切相关，可能参与调节肿瘤细胞的骨架重建、细胞黏附性和上皮间充质化的过程，这可能成为治疗多种类型肿瘤的药物靶点。本文对PRL-3与肿瘤关系的研究现状和进展做一综述。展开更多
文摘Background Considerable evidence suggests that phosphatase of regenerating liver-3 （PRL-3） plays multiple roles in cancer metastasis; however, the molecular mechanisms remain largely unknown. The aim of this study was to identify proteins associated with PRL-3-promoted colon cancer metastasis, by comparative proteomic analysis. Methods Proteomes of human colon cancer LoVo cells transfected with PRL-3 gene （LoVo-PRL-3） or empty vector PAcGFP-C3 （LoVo-control） were compared using 2D gel electrophoresis. Proteins that varied significantly in concentration were selected and identified using mass spectrometry. Expression of translationally controlled tumor protein （TCTP） mRNA and protein in LoVo-PRL-3 and LoVo-control cells was detected by real-time PCR and Western blotting. Small interfering RNA （siRNA） targeting TCTP was used for silencing TCTP expression in LoVo-PRL-3 cells. Functional significance of TCTP in PRL-3-promoted colon cancer cell proliferation, migration and invasion was investigated by Cell Counting Kit-8 assay and transwell chamber. Results Seventeen proteins displaying significant and reproducible differences between LoVo-PRL-3 and LoVo-control cells were identified. Ten proteins were upregulated and seven were downregulated in LoVo-PRL-3 cells when compared with LoVo-control cells. Eight identified proteins are associated with distinct steps of tumor metastasis： ubiquitin-like protein ISG15, interleukin-18, TCTP, serpin B5, annexin A3, macrophage-capping protein, ATP-dependent RNA helicase DDX3X, and cathepsin D. Real-time PCR and Western blotting results showed that both TCTP mRNA and protein were significantly increased in LoVo-PRL-3 cells compared to LoVo-control cells. Transfection with TCTP siRNA significantly reduced the expression of both mRNA and protein levels of TCTP in LoVo-PRL-3 cells. Knockdown of TCTP by siRNA inhibited PRL-3-promoted proliferation, migration and invasion of LoVo-PRL-3 cells. Conclusion Our results imply that TCTP might be a mediator of PRL-3-promoted proliferation, migration and invasion of human colon cancer cells.
基金Supported by Grant for Key Technology Research and Development Program 2002BA711A06National Basic Research Priorities Program 973 Project 1998051203the Ministry of Science and Technology of China,and grant H020920030390,from the Beijing Science and Technology Commission
文摘AIM:To investigate the prognostic significance of phosphatase regenerating liver 3(PRL-3)protein expression in gastric cancer.METHODS:PRL-3 expression in paraffin-embedded tumor specimens from 293 patients with gastric cancer was studied retrospectively by immunohistochemistry.Monoclonal antibody specifically against PRL-3,3B6,was obtained with hybridoma technique.RESULTS:Positive PRL-3 expression was detected in 43.3%(127 of 293)of gastric cancer cases.High expression of PRL-3 was positively correlated with tumor size,depth of invasion,vascular/lymphatic invasion,lymph node metastasis,high TNM stage and tumor recurrence.Patients with positive PRL-3 expression had a significantly lower 5-year survival rate than those with negative expression(28.3%vs 51.9%,P<0.0001).Patients who received curative surgery,and with positive PRL-3 expression had a significant shorter overall survival and disease-free disadvantage over patients with negative expression(hazard ratio of 16.7 and 16.6,respectively;P<0.0001 for both).Multivariate analysis revealed that PRL-3 expression was an independent prognostic indicator for overall and disease-free survival of gastric cancer patients,particularly for survival in TNM stageⅢpatients.CONCLUSION:PRL-3 expression is a new independent prognostic indicator to predict the potential of recurrence and survival in patients with gastric cancer at the time of tumor resection.
文摘目的:研究miR495、miR551a干扰质粒对SGC7901胃癌细胞中促肝细胞再生磷酸酶-3(phosphatase of regenerating liver-3,PRL-3)表达的影响,探讨基因干扰在胃癌治疗中的价值.方法:分别体外培养稳定转染后的SGC7901细胞系及未经处理的SGC7901细胞系,取对数生长期瘤细胞,0.5mL(1×107/mL)细胞悬液接种于Balb/ca(nu/nu)裸鼠腹腔内,在SPF条件下饲养1mo后处死,观察裸鼠成瘤率、瘤体生长情况及腹膜转移等情况,并行组织病理学检查.取各组部分移植瘤行荧光定量PCR检测,对比各组miR495、miR551a及PRL-3mRNA的相对表达水平.结果:各组裸鼠成瘤率均100%,其中两实验组裸鼠一般情况及生存期均好于对照组.取各组移植瘤行荧光定量PCR检测显示,两实验组裸鼠miRNA表达量显著高于对照组,PRL-3mRNA表达量低于对照组.转染质粒组胃癌迁移能力明显减弱.结论:转染靶向干扰PRL-3表达的miR495、mi551a真核质粒可以明显抑制胃癌细胞体内转移侵袭能力.
文摘胃癌是世界范围内常见肿瘤之一,因胃癌死亡的患者数占据肿瘤相关死因的第3位.肝再生磷酸酶-3(phosphatase of regenerating liver 3,PRL-3)作为一个新近发现的蛋白酪氨酸磷酸酶,近年来研究发现其在胃癌组织中高表达,在胃癌淋巴转移、腹膜转移等发挥重要作用,与胃癌患者预后负相关.其后,越来越多的研究关注其在胃癌发生、发展中的调控机制,以期阐明PRL-3在胃癌发生、发展中的具体调节通路及影响因素.尽管随着研究的不断深入,对于PRL-3在胃癌中的作用机制得到一部分阐释,但对于P R L-3在促进胃癌淋巴转移、腹膜转移等恶性进展及复发的机制仍然不是很清楚,本文即对近年来PRL-3的相关研究进展作一综述.
文摘目的观察慢病毒介导的sh RNA沉默肝再生磷酸酶-3(PRL-3)基因对结肠癌SW480细胞增殖、侵袭、凋亡的影响。方法实验分组为空白对照组、阴性对照组、转染组。将携带PRL-3 sh RNA的慢病毒载体转染结肠癌SW480细胞,建立稳定沉默PRL-3的细胞株,real-time PCR检测转染后PRL-3 m RNA的相对表达水平。采用MTT法、平板克隆形成实验检测转染后细胞增殖能力;采用Transwell侵袭实验、侵袭小室法检测转染后细胞迁移及侵袭能力;采用流式细胞术检测转染后细胞凋亡率变化。结果稳定沉默PRL-3的细胞株构建成功,转染组PRL-3m RNA的相对表达水平低于空白对照组、阴性对照组(P<0.05),空白对照组、阴性对照组比较差异无统计学意义。PRL-3 sh RNA转染SW480细胞72 h后,转染组与空白对照组、阴性对照组比较,细胞增殖能力受到抑制,转染120 h时最明显(P<0.05)。转染组克隆形成能力较空白对照组、阴性对照组下降(P<0.05)。转染组与空白对照组、阴性对照组比较,细胞迁移、侵袭能力下降,凋亡率增加(P<0.05)。结论结肠癌SW480细胞转染PRL-3 sh RNA可减少PRL-3的表达,有效抑制SW480细胞增殖,促进其凋亡,PRL-3可能成为治疗结肠癌的靶基因。
文摘目的:观察促肝再生磷酸酶-3(phosphatase of regeneration liver-3,PRL-3)抑制剂原钒酸钠(sodium orthovanadate,SoV)对大肠癌细胞株Colo-320迁移能力的影响.方法:应用Westernblot方法检测PRL-3在7株大肠癌细胞中的表达,筛选表达最强的1株做下一步抑制实验.选择PRL-3拮抗剂SoV,通过细胞划痕实验观察其在0.5μmol/L浓度下对癌细胞运动能力的影响,高倍倒置显微镜下计算细胞迁移距离.细胞爬片原位杂交观察SoV对PRL-3mRNA的表达.结果:PRL-3表达最强的结肠癌细胞株为Colo-320.细胞划痕实验显示,SoV作用48h细胞仅迁移相当于4-7个细胞的距离,400×倍镜下精确测量20个细胞在0-48h迁移距离,计算对照组细胞迁移速度为39.12±10.11μm/h,SoV为12.84±6.78μm/h,差异非常显著(P<0.00001).SoV作用细胞48h后原位杂交结果未见PRL-3mRNA阳性表达.结论:SoV能显著抑制Colo-320细胞迁移能力,其机制可能与抑制PRL-3酶活性以及基因转录有关.
文摘肝再生磷酸酶-3（PRL-3）属于蛋白酪氨酸磷酸酶家族，它发挥作用的方式是通过调节酪氨酸残基的磷酸化状态实现对蛋白质活性的调控。已有的研究表明，P R L-3与人多种肿瘤的发生、发展、转移及预后密切相关，可能参与调节肿瘤细胞的骨架重建、细胞黏附性和上皮间充质化的过程，这可能成为治疗多种类型肿瘤的药物靶点。本文对PRL-3与肿瘤关系的研究现状和进展做一综述。
