肝再生磷酸酶-3在胃癌中的研究进展

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.     

RNAi沉默促肝细胞再生磷酸酶-3基因抑制肝癌细胞侵袭

促肝细胞再生磷酸酶(PRL)-3基因是PRLs家族重要成员之一,与某些肿瘤侵袭转移有关[1].最近研究发现,PRL-3基因在肝癌细胞中高表达[2],但目前尚不清楚PRL-3基因在肝癌细胞侵袭转移中的可能作用和机制.     

大黄素对卵巢癌HO-8910PM细胞中PRL-3和NAG-1表达的影响

目的 研究大黄素对人高转移卵巢癌HO-8910PM细胞中促肝细胞再生磷酸酶-3(PRL-3)和非类固醇抗炎药物激活基因(NAG-1)表达的影响.方法 采用Western blot法检测大黄素对HO-8910PM细胞中PRL-3和NAG-1蛋白表达的影响,荧光定量实时PCR分析大黄素对HO-8910PM细胞中PRL-3和NAG-1 mRNA表达的影响.结果 大黄素能明显下调HO-8910PM细胞中PRL-3的表达,强烈上调NAG-1表达.结论 大黄素影响高转移卵巢癌细胞转移能力可能与PRL-3和NAG-1表达有关.     

PRL-3真核绿色荧光蛋白表达载体的构建及其在结肠癌SW480细胞中的表达

目的:构建蛋白酪氨酸磷酸酶-3(PRL-3)绿色荧光蛋白真核表达载体pEGFP-N1-PRL-3,为PRL-3基因功能研究奠定基础.方法:设计人PRL-3特异性引物,提取人大肠癌细胞系SW620细胞总RNA,应用RT-PCR方法获取人PRL-3全长cDNA,分别克隆至T载体及真核绿色荧光蛋白表达载体pEGFP-N1,应用PCR、酶切及DNA测序进行鉴定,确认后转染大肠癌细胞SW480,应用G418进行筛选获取PRL-3稳定表达细胞克隆,应用荧光定量PCR检测PRL-3基因表达.结果:获得522 bp的PRL-3基因编码序列并成功构建了真核绿色荧光蛋白表达载体pEGFPN1-PRL-3,该重组载体能够在SW480细胞中稳定表达.结论:成功构建真核绿色荧光蛋白表达载体pEGFP-N1-PRL-3和建立PRL-3稳定表达SW480细胞,为进一步深入研究PRL-3基因在大肠癌发生发展中的作用奠定基础.     

PRL-3、MMP-2和CD105在食管鳞癌中的表达及其与临床病理特征的关系

目的探讨蛋白酪氨酸磷酸酶(PRL)-3、基因金属蛋白酶(MMP)-2和细胞膜糖蛋白(CD105)在食管鳞状细胞癌中的表达及其与食管鳞癌临床病理特征之间的关系。方法选取2012年12月至2014年12月在该院收集的食管鳞癌患者手术组织细胞标本为研究组,同时取该组患者癌旁组织细胞标本为对照组,应用免疫组化SP检测PRL-3蛋白、MMP-2蛋白和CD105-MVD的表达,并计算PRL-3蛋白、MMP-2蛋白和CD105-MVD阳性表达与食管鳞癌临床病理学参数之间的关系。结果 PRL-3、MMP-2和CD105-MVD在食管鳞状细胞癌中的阳性表达率显著高于癌旁组织(P0.05)。MMP-2和CD105-MVD与食管鳞癌患者的分期、淋巴结转移、浸润程度有相关性,而PRL-3与食管鳞癌患者的淋巴结转移有显著相关性。结论 PRL-3、MMP-2和CD105-MVD在食管鳞状细胞癌中高表达,提示三者联合检测或可以筛选成为食管鳞状细胞癌诊断的早期标志物,而MMP-2和CD105可以作为预测食管鳞癌侵袭转移和评估预后的生物标志物。     

磷酸酶SHP-1与消化系统肿瘤关系的研究现状

含有SH2结构域的蛋白酪氨酸磷酸酶SHP-1,属于非受体型蛋白酪氨酸磷酸酶家族,主要在造血细胞和上皮细胞中表达,是多种信号转导通路如JAK/STAT、PI3K-AKT等的负向调节因子.SHP-1在抑制淋巴瘤、白血病等肿瘤的发生及发展中起到重要作用.随着生物技术的进步及SHP-1基因突变小鼠模型被广泛应用,SHP-1在消化系统肿瘤(特别是肝癌)的发生、发展方面所起的作用也逐渐被发现.     

子宫腺肌病组织中PRL-3、VEGF的表达变化及意义

目的 观察子宫腺肌病（AM）组织中促肝细胞再生磷酸酶-3（PRL-3）及血管内皮细胞生长因子（VEGF）的表达变化,并探讨其意义.方法 选取30例AM患者异位内膜（异位组）、在位内膜组织（在位组）,以及30例子宫肌瘤患者正常子宫内膜组织（对照组）,采用免疫组化法检测各组织中PRL-3、VEGF的表达水平.结果 VEGF及PRL-3在异位组表达显著高于在位组及对照组（P均＜0.01）,且在在位组的表达高于对照组（P均＜0.05）.异位组、在位组增生期与分泌期内膜中PRL-3的表达均高于同时相对照组（P均＜0.05）.对照组中VEGF在分泌期内膜中的表达高于增生期（P＜0.05）.PRL-3与VEGF在在位组中的表达呈正相关关系（r=0.439,P＜0.05）.结论 PRL-3及VEGF在AM异位内膜中呈高表达,二者可能与AM的发病有关.     

抑制促肝再生磷酸酶-3对大肠癌细胞迁移能力的影响

目的:观察促肝再生磷酸酶-3(phosphatase ofregeneration 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Western blot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0.5 μmol/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-3 mRNA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48 h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0-48 h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±6.78 μm/h,差异非常显著(P<0.00001).SoV作用细胞48 h后原位杂交结果未见PRL-3 mRNA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.     

RNAi沉默PRL-3基因对大肠癌细胞侵袭的抑制

目的:探讨RNA干扰(RNA interference,RNAi)沉默促肝细胞再生磷酸酶3(proteins in regenerating liver cell-3,PRL-3)基因对大肠癌细胞侵袭的影响.方法:应用PRL-3基因小干扰RNA(small interfering RNA,siRNA)转染处理大肠癌细胞系HCT116后,采用荧光实时定量PCR方法检测PRL-3基因mRNA水平,分别采用软琼脂集落培养实验和Boyden小室模型实验检测癌细胞的锚着不依赖性增殖和侵袭能力.其次将转染48 h的细胞接种裸鼠,观察对癌细胞体内侵袭的影响.结果:与两对照组比较,siRNA组PRL-3mRNA水平明显降低,且呈浓度和时间依赖性(P＜0.05).体外实验发现:与对照组比较,PRL-3 siRNA转染组软琼脂集落形成数和穿过滤膜的癌细胞均呈剂量依赖性减少(P＜0.05,P＜0.05).体内实验发现:空白对照组和空载对照组有较多癌细胞侵袭癌肿组织周围的横纹肌,并侵犯血管,而siRNA转染组未见这些现象.结论:PRL-3在大肠癌细胞侵袭中起着重要的作用,采用PRL-3 siRNA转染可抑制大肠癌细胞侵袭.     

肝再生磷酸酶3对SGC7901细胞迁移侵袭及RhoC表达的影响

目的:通过观察肝再生磷酸酶3(phosphatase of regenerating liver 3,PRL-3)对胃癌SGC7901细胞迁移侵袭及RhoC表达的影响,探讨胃癌SGC7901细胞迁移侵袭的机制中PRL-3和RhoC是否位于同一信号通路.方法:体外培养人胃癌SGC7901细胞,经不同浓度PRL-3抗体(1∶600,1∶400,1∶200)作用后,采用细胞划痕实验观察不同时间段(0、12、24、48 h)SGC7901细胞的迁移距离;通过Transwell迁移侵袭实验检测不同浓度PRL-3抗体(1∶600,1∶400,1∶200)作用48 h后SGC7901细胞迁移侵袭的穿膜细胞数,并采用Real-time PCR和ELISA检测SGC7901细胞中RhoC mRNA及蛋白表达量的变化,比较不同浓度PRL-3抗体处理后SGC7901细胞迁移侵袭的细胞数和RhoC mRNA及蛋白的表达量.结果:与对照组比较,随着PRL-3抗体浓度的升高和作用时间的延长,SGC7901细胞的迁移距离逐渐减小,在48 h,经不同浓度PRL-3抗体(1∶600,1∶400,1∶200)处理SGC7901细胞后,其迁移和侵袭的细胞数,RhoCmRNA相对表达量和蛋白表达水平与对照组相比均明显降低(迁移SGC7901细胞数:365.0±5.0,165.3±5.0,90.3±5.5 vs 512.3±4.9;侵袭SGC7901细胞数:321.3±6.1,179.0±6.1,75.7±4.0 vs 545.3±5.0;RhoC mRNA的相对表达量:0.910±0.022,0.742±0.018,0.539±0.015 vs 1.000±0.000;RhoC蛋白表达量:1130.77 g/mL±15.32 g/mL,981.52 g/mL±14.44 g/mL,893.03 g/mL±11.10 g/mL vs 1212.42 g/mL±18.37g/mL)(均P0.01),不同PRL-3抗体浓度组两两比较均有统计学意义(均P0.01).结论:PRL-3在体外能显著促进SGC7901细胞的迁移侵袭,并能增强RhoC基因和蛋白的表达,提示在SGC7901细胞迁移侵袭的机制中PRL-3和RhoC可能位于同一信号通路.     

The association of the expression level of protein tyrosine phosphatase PRL-3 protein with liver metastasis and prognosis of patients with colorectal cancer

PURPOSE: To investigate PRL-3 protein expression in normal colorectal epithelia and colorectal cancers with monoclonal antibody (MAb) against PRL-3. METHODS: MAb against PRL-3 was prepared with the hybridoma technique, and its specificity was confirmed with ELISA and Western blotting assays. The expression of PRL-3 protein in normal colorectal epithelia and colorectal cancers was examined by immunohistochemistry assay. Logistic regression and survival analysis were performed to determine the clinical significance of PRL-3 expression. RESULTS: MAb 3B6 against PRL-3 was obtained and showed high specificity. PRL-3 protein was expressed in two of 28 (7.1%) normal colorectal epithelia, 21 of 88 (23.9%) primary colorectal cancers, 22 of 41 (53.7%) metastatic lymph nodes and eight of 12 (66.7%) liver metastases, respectively. The PRL-3 expression rates of metastases were significantly higher than those of primary colorectal cancers and normal colorectal epithelia (P < 0.05). PRL-3 expression was significantly associated with the liver metastasis of colorectal cancer (P = 0.004) and tended to shorten survival time (P = 0.0145). CONCLUSIONS: This is the first study demonstrating that PRL-3 is a potential marker for liver metastasis of colorectal cancer and negatively influences the prognosis of colorectal cancer patients.     

Overexpression and involvement in migration by the metastasis-associated phosphatase PRL-3 in human myeloma cells

Multiple myeloma (MM) is characterized by accumulation and dissemination of malignant plasma cells (PCs) in the bone marrow (BM). Gene expression profiling of 2 MM cell lines (OH-2 and IH-1) indicated that expression of PRL-3, a metastasis-associated tyrosine phosphatase, was induced by several mitogenic cytokines. Cytokine-driven PRL-3 expression could be shown in several myeloma cell lines at both the mRNA and protein levels. There was significantly higher expression of the PRL-3 gene in PCs from patients with monoclonal gammopathy of undetermined significance (MGUS), smoldering myeloma (SMM), and myeloma than in PCs from healthy persons. Among 7 MM subgroups identified by unsupervised hierarchical cluster analysis, PRL-3 gene expression was significantly higher in the 3 groups denoted as "proliferation," "low bone disease," and "MMSET/FGFR3." PRL-3 protein was detected in 18 of 20 BM biopsies from patients with MM. Silencing of the PRL-3 gene by siRNA reduced cell migration in the MM cell line INA-6, but had no detectable effect on proliferation and cell-cycle phase distribution of the cells. In conclusion, PRL-3 is a gene product specifically expressed in malignant plasma cells and may have a role in migration of these cells.     

Expression of phosphatase regenerating liver 3 is an independent prognostic indicator for gastric cancer

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. Nonoclonal antibody specifically against PRL-3, 3B6, was obtained with hybridoma technique.
RESULTS： Positive PRL-3 expression was detected in 43.3% （227 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 52.9%, 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 stage Ⅲ patients.
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,     

PRL-2 increases Epo and IL-3 responses in hematopoietic cells

Dual specificity protein tyrosine phosphatase PRL-2 is overexpressed in pediatric acute myeloid leukemia (AML) and is located at human chromosome 1p35, a region often rearranged or amplified in malignant lymphoma and B-cell chronic lymphocytic leukemia (B-CLL). Little is known of the significance of PRL-2 expression in hematopoietic malignancies. Herein we demonstrated that ectopic expression of PRL-2 in murine pre-B-cell line Baf3ER and mouse bone marrow cells induced key features associated with malignant progression and metastasis. PRL-2-transfected Baf3ER cells had augmented growth responses to hematopoietic growth factors Epo or IL-3 with shortened cell cycle, reduced requirement (5×) for Epo in cell survival, increased cell migration (3×), reduced cell adhesion (5×), and conversion to an immature cell morphology in association with increased expression (3×) of stem cell marker Bmi-1. When transduced into mouse bone marrow cells, PRL-2 increased Epo-induced colony formation (4×) and gave rise to larger colonies. These observations provide evidences implicating PRL-2 as a pathogenic molecule in hematopoietic malignancies and suggest its potential as a novel therapeutic target.     

Elevated PRL-3 expression was more frequently detected in the large primary gastric cancer and exhibits a poor prognostic impact on the patients

Purpose  It has been reported that elevated PRL-3 expression was closely associated with lymph node metastasis of gastric cancer. In the present study, we detected the expression of PRL-3 in primary gastric cancer tissue, and evaluated its prognostic impact on the patients. Methods  Total 137 gastric tumor samples were measured for PRL-3 phosphatase expression using immunohistochemistry method, and the overall survival rate was compared between the patients with high PRL-3 expression (n = 85) and those with moderate or low PRL-3 expression (n = 52) in the primary tumor. Results  PRL-3 expression was more frequently detected in the tumors with a diameter >40 mm and in advanced stages (TNM stage III or IV). Furthermore, the overall survival rate of the patients with high expression of PRL-3 in the primary tumor was significantly less than those with moderate or low expression (P < 0.001). Conclusions  PRL-3 expression is associated with gastric cancer progression. Its high expression in the primary lesion had a negative impact on the prognosis of the patients. This strongly suggests that PRL-3 should be considered as a prognostic factor. Zhao Wang and Shi-Rong Cai have contributed equally to this work.     

PRL-3基因对结直肠癌细胞增殖能力的影响

目的:探讨PRL-3的过表达或敲低对结直肠癌细胞增殖能力的影响.方法:利用MTT法、平板克隆形成实验检测PRL-3对细胞体外增殖的影响;应用流式细胞术检测PRL-3对细胞周期的影响.结果:应用MTT法,检测PRL-3对SW480/EGFP、SW480-EGFP-PRL-3、SW480/EGFP/Mock及SW480-PRL-3-KD1细胞体外增殖能力的影响,经析因方差分析,4组差异具有显著性(F=23.463,P=0.000);不同时间点对细胞体外增殖的影响差异具有显著性(F=71.515,P=0.000);各组细胞与各时间组两因素交互效应显著(F=2.128,P=0.008);除第1天外,其他各时间点细胞组间的细胞增殖差异具有显著性.经LSD法多重比较,结果表明,与SW480/EGFP/Mock和SW480/EGFP细胞相比,SW480-EGFP-PRL-3细胞的增殖速度加快,而SW480-PRL-3-KD1细胞的增殖速度减慢.平板克隆形成实验显示SW480-EGFP-PRL-3细胞克隆形成能力明显增强,而SW480-PRL-3-KD1细胞克隆形成能力显著下降,差异具有显著的统计学意义(F=44.411,P=0.000).结论:PRL-3基因可促进结直肠癌细胞的增殖.     

A retrospective cohort study of clinical value of PRL-3 in stage III human colorectal cancer

The aim of this study was to investigate the expression of phosphatase of regenerating live-3 (PRL-3) in human stage III colorectal cancer (CRC) and to evaluate its correlation with metachronous liver metastasis (MLM) and prognosis.The retrospective cohort study included 116 stage III CRC primary tumors and 60 normal colorectal tissues. PRL-3 expression was measured by immunohistochemistry. We investigated the correlation of PRL-3 with clinicopathologic features by the chi-square test. The association of PRL-3 expression with MLM was assessed by binary logistic regression. Overall survival (OS) and disease-free survival (DFS) between patients with positive PRL-3 expression and those with negative PRL-3 expression were compared by the Kaplan–Meier method and Cox proportional hazards regression model.We found that 32.8% of stage III CRC primary tumors were PRL-3 positive, and 15.0% of normal colorectal epithelia showed high PRL-3 expression (P = .012). Seventeen tumors (47.2%) among 36 cases that developed MLM were PRL-3 positive, and only 21 tumors (26.3%) in the 80 cases that did not develop MLM had positive PRL-3 expression (P = .026). PRL-3 expression was associated with MLM (P = .028). Patients with positive expression of PRL-3 showed a significantly shorter OS (40.32 ± 3.97 vs 53.96 ± 2.77 months, P = .009) and DFS (34.97 ± 4.30 vs 44.48 ± 2.89 months, P = .036). A multivariate analysis indicated that PRL-3 expression was an independent unfavorable prognostic factor for OS (P = .007).Our study suggested that high PRL-3 expression is an independent risk factor for MLM and poor prognosis. PRL-3 is expected to be a promising biomarker for predicting the incidence of MLM and prognosis in patients with stage III CRC.     

Combined phenotype of 4 markers improves prognostic value of patients with colon cancer

IntroductionCombination of multiple biomarkers representing distinct aspects of tumor biology will have a better prognostic value. This study was to identify prognostic subgroups of colon adenocarcinoma by combined analysis of synuclein-gamma (SNCG), a human homologue of piwi (Hiwi), phosphatase of regenerating liver-3 (PRL-3), arrest-defective protein 1, homolog A (ARD1) and clinicopathologic features in 225 colon adenocarcinoma specimens.MethodsImmunohistochemistry for 4 tumor markers was performed in whole tissue sections from 225 colon adenocarcinoma patients with complete clinicopathologic data and up to 10-year follow-up. The immunohistochemical expression patterns were examined individually and in multimarker combinations. Univariate and multivariate analyses were performed to identify independent predictive markers of poor outcome.ResultsWith the tumor marker positive rate [32.0% (62/225) for SNCG; 76.9% (173/225) for combined SNCG/Hiwi/PRL-3/ARD1] and the detecting accuracy [61.9% (252/407) for SNCG; 82.6% (336/407) for combined SNCG/Hiwi/PRL-3/ARD1] increasing, incremental value of combined SNCG/Hiwi/PRL-3/ARD1 (P < 0.001; hazard ratios (HR), 3.2) to poor outcome was found. Stratified by lymph node, Hiwi alone (P = 0.004; HR, 3.2) led to poor outcome in patients without lymph node metastasis (LN—), and SNCG (P < 0.001; HR, 2.5) had independently poor prognostic value for patients with lymph node metastasis (LN +).ConclusionsMultimarker phenotypes improved tumor positive rate, detecting accuracy and prognostic value. In addition, a subgroup of more aggressive tumors can be identified by evaluating Hiwi level in LN— cancer, and SNCG level in LN + cancer.     

Prognostic and metastatic value of phosphatase of regenerating liver-3 in invasive breast cancer

Purpose  

The aim of this study was to investigate the expression of the PRL-3 in human invasive breast cancer and to evaluate its clinical and prognostic significance. Its potential role in the invasive-metastatic properties of invasive breast cancer was also investigated.     

Two fragments from fibrinolysin digests of ovine prolactin: characterization and recombination to generate full immunoreactivity.

The action of fibrinolysin (plasmin; EC 3.4.21.7) on ovine prolactin has been investigated. It was found that the enzyme selectively cleaves the bond between Met-53 and Ala-54. The two fragments, PRL-(1-53) and PRL-(54-199), have been purified and characterized. A recombinant molecule has been obtained by noncovalent interaction of PRL-(1-53) and PRL-(54-199). The recombined protein behaves nearly identically to the parent hormone in circular dichroism spectra and exclusion chromatography. The recombinant possesses full immunoreactivity, as revealed by gel double-diffusion and complement fixation. However, the recombined protein exhibits low prolactin activity in the pigeon crop-sac test.