DEAH-box polypeptide 32 promotes hepatocellular carcinoma progression via activating the β-catenin pathway

PurposeHepatocellular carcinoma (HCC) is refractory cancer with high morbidity and high mortality. DEAH-box polypeptide 32 (DHX32) was upregulated in several types of malignancies and predicted poor prognosis. Herein, we investigated the role of DHX32 in HCC progression.MethodsThe expression of DHX32, β-catenin, and epithelial-mesenchymal transition (EMT)-related makers were determined by Western blot and quantitative real-time PCR assays. Cell proliferation was tested by EdU cell proliferation assay. The effect of DHX32 and β-catenin on cell migration and invasion were detected by wound-healing and Traswell invasion assays. Tumour xenografts were performed to determine the effect of DHX32 on HCC tumour growth.ResultsHigh level of DHX32 expression was associated with reduced overall survival in HCC patients. DHX32 expression was upregulated in human HCC cells and ectopic expression of DHX32 induced EMT, promoted the mobility and proliferation of HCC cells, and enhanced tumour growth in vivo. Silencing DHX32 reversed EMT, inhibited the malignancy behaviors of HCC cells, and suppressed tumour growth. Mechanistically, silencing DHX32 decreased the expression of β-cateninin in nucleus and β-catenin siRNA abrogated DHX32-mediated HCC progression.ConclusionDHX32 was an attractive regulator of HCC progression and indicated DHX32 canserve as a potential biomarker and therapeutic target for HCC patients.     

CircRNA WHSC1 promotes non‐small cell lung cancer progression via sponging microRNA‐296‐3p and up‐regulating expression of AKT serine/threonine kinase 3

BackgroundLung cancer is the most commonly diagnosed cancer and leading cause of cancer death, with 80%–85% of non‐small cell lung cancer (NSCLC). Circular RNAs (circRNAs) have been shown to be promising early diagnostic and therapeutic molecular biomarkers for NSCLC. However, biological role and regulatory mechanism of circRNA WHSC1 (circWHSC1) in NSCLC are unknown. Therefore, we aim to explore the function and mechanism of circWHSC1 in NSCLC oncogenesis and progression.MethodsqRT‐PCR was used for circWHSC1 level evaluation; Kaplan‐Meier was used for survival analysis; bioinformatics, dual‐luciferase activity, and RNA pull‐down were used for evaluating competing endogenous RNA (ceRNA) network; cell viability, colony formation, apoptosis, migration, and invasion were used for cell function analysis; function gain and loss with rescue experiments were used for exploring mechanism of circWHSC1 in NSCLC development.ResultsSignificantly up‐regulated circWHSC1 and down‐regulated microRNA‐296‐3p (miR‐296‐3p) were identified in NSCLC tissues and cells. Up‐regulated circWHSC1 was associated with poor prognosis in NSCLC patients. MiR‐296‐3p was sponged by circWHSC1, and AKT serine/threonine kinase 3 (AKT3) was target of miR‐296‐3p; meanwhile, miR‐296‐3p over‐expression significantly down‐regulated AKT3 expression, and co‐transfecting anti‐miR‐296‐3p rescued circWHSC1 silence caused AKT3 down‐regulation. CircWHSC1 silence significantly inhibited colony formation, viability, invasion, and migration, while increased NSCLC cell apoptosis, which were partially rescued by anti‐miR‐296‐3p.ConclusionCircWHSC1 is an independent indicator of poor prognosis in NSCLC patients, and functions as a ceRNA of miR‐296‐3p to up‐regulate AKT3, consequently promotes NSCLC cell growth and metastasis. Targeting circWHSC1 might be a prospective strategy for diagnosis, therapeutics, and prognosis of NSCLC.     

Curcumin ameliorates epithelial-to-mesenchymal transition of podocytes in vivo and in vitro via regulating caveolin-1

AimsEpithelial-mesenchymal transition (EMT) is recognized to play a key role in diabetic nephropathy (DN). Curcumin, the main active component of turmeric extracted from the roots of the Curcuma longa plant, has been reported for its anti-fibrotic effects in kidney fibrosis. The purpose of our study was to investigate the effects of curcumin in reversing epithelial-to-mesenchymal transition (EMT) of podocytes in vivo and in vitro.Materials/methodsIn vivo streptozotocin (STZ)-induced diabetic rats received vehicle or curcumin, and podocytes were treated with high glucose (HG) in the presence or absence of curcumin in vitro. And we investigated the effect of curcumin on HG-induced phosphorylation of cav-1 on the stability cav-1 and β-catenin using immunoprecipitation and fluorescence microscopy analysis.ResultsCurcumin treatment dramatically ameliorated metabolic parameters, renal function, morphological parameters in diabetic rats. We found that HG treatment led to significant down-regulation of p-cadherin and synaptopodin, as well as remarkable up-regulation of α-SMA and FSP-1 in vivo and in vitro. Furthermore, curcumin inhibited HG-induced caveolin-1 (cav-1) Tyr¹⁴ phosphorylation associating with the suppression of stabilization of cav-1 and β-catenin.ConclusionsIn summary, these findings suggest that curcumin prevents EMT of podocytes, proteinuria, and kidney injury in DN by suppressing the phosphorylation of cav-1, and increasing stabilization of cav-1 and β-catenin.     

肾细胞癌中Wif-1基因的甲基化状态及表达

目的检测肾细胞癌(RCC)中Wnt抑制因子-1(Wif-1)基因的甲基化状态及表达水平,探讨其在肾细胞癌发生发展中的可能机制。方法分别应用甲基化特异性PCR(MSP)和逆转录-聚合酶链式反应(RT-PCR)检测56例肾细胞癌及相应癌旁正常肾组织中Wif-1基因的甲基化状态及表达水平。结果 Wif-1基因在RCC组织中的甲基化率显著高于相应癌旁正常肾组织(P<0.05)。Wif-1基因在RCC组织中的相对表达量显著低于相应癌旁正常肾组织(P<0.05)。在RCC组织中,Wif-1基因甲基化阳性组mRNA的相对表达量与甲基化阴性组mRNA的相对表达量比较无显著差异(P>0.05)。结论 Wif-1基因启动子区的高甲基化在肾细胞癌的发生中是一个频发事件。     

肾透明细胞癌组织中B7-H3的表达及意义

目的评价B7-H3在肾透明细胞癌组织中的表达及其与预后的关系。方法采用免疫组化方法检测154例肾透明细胞癌患者肿瘤标本中B7-H3的表达,分析其与患者临床病理因素以及术后生存时间之间的关系。结果 154例患者中,B7-H3阳性28例（18.18%）,其中无瘤生存6例,复发7例,死亡15例,至最后随访日期,总生存期（OS）为（19.71±20.32）个月;B7-H3阴性126例（81.82%）,其中无瘤生存119例,复发3例,死亡4例,OS为（33.47±18.32）个月,两者比较差异有统计学意义（P〈0.05）。单因素分析表明,B7-H3阳性表达患者在肿瘤大小、原发肿瘤分期、区域淋巴结转移、远处转移、临床分期等方面较阴性表达患者有明显差异,B7-H3阳性表达患者术后生存时间较阴性表达患者明显缩短。结论 B7-H3的表达可能与肾透明细胞癌的转移和进展有关,可以作为肾透明细胞癌的一个独立预后因素。     

MicroRNA-199a-3p在肾癌中的表达及作用

目的检测microRNA-199a-3p（miR-199a-3p）在肾癌细胞株和组织中的表达情况并探究miR-199a-3p在肾癌细胞中的作用。方法利用实时定量RT-PCR检测miR-199a-3p在肾癌细胞和组织中的表达水平;利用miR-199a-3p模拟物转染肾癌细胞786-0上调miR-199a-3p后,通过CCK-8、克隆形成、Transwell以及细胞周期检测来探究其在肾癌细胞中的作用。结果 miR-199a-3p在肾癌细胞中明显低表达,在78%（14/18）的肾癌组织中亦明显低表达;上调miR-199a-3p可显著抑制肾癌细胞的增殖、存活和侵袭并能诱导细胞周期G1期阻滞。结论我们的研究显示在肾癌中miR-199a-3p明显低表达并参与肾癌的发生、发展,这表明miR-199a-3p具有作为肾癌诊断和治疗靶点的潜能。     

肾透明细胞癌组织中B7-H1的表达及其与预后的关系

目的探讨B7-H1在肾透明细胞癌组织中的表达及其与预后的关系。方法采用免疫组织化学方法检测132例肾透明细胞癌患者术后肿瘤标本中B7-H1的表达,单因素及多因素分析B7-H1的表达与预后的关系。结果 132例患者中,B7-H1阳性46例（34.85%）,其中无瘤生存26例,复发6例,死亡14例,至最后随访期限,疾病无进展生存期（PFS）为（32.29±12.56）个月,总生存期（OS）为（27.08±15.43）个月;阴性86例（65.15%）,其中无瘤生存79例,复发3例,死亡4例,PFS为（38.77±20.00）个月,OS为（37.02±20.00）个月,两者比较差异有统计学意义（P〈0.05）。单因素分析表明,B7-H1阳性表达与肿瘤大小、原发肿瘤分期、区域淋巴结转移、远处转移、临床分期、组织学分级呈显著正相关（均P〈0.05）,而与患者年龄、性别无相关性。多因素分析表明,在控制肿瘤大小、TNM分期、临床分期、组织学分级的影响后,B7-H1阳性表达具有更高的肿瘤特异性死亡率[危险因素（RR）3.918,95%可信区间（95%CI）1.305～11.765,P〈0.05]和复发率[RR4.486,95%CI1.404～14.333,P〈0.05]。结论 B7-H1阳性表达与肾透明细胞癌患者术后死亡率及复发率呈正相关,B7-H1是肾透明细胞癌的一个独立预后因素。     

Claudin-1、Id-1、Id-3蛋白在胃癌中的表达及临床意义

目的研究胃癌组织Claudin-1、Id-1、Id-3表达与胃癌细胞分化、增殖、侵袭转移和预后的关系。方法应用免疫组织化学检测Claudin-1、Id-1、Id3-蛋白在胃癌组织中的表达。结果Claudin-1表达水平与胃癌组织学类型、肿瘤侵袭程度及淋巴结转移相关（P〈0．05）,Id-1蛋白表达与胃癌淋巴结转移呈负相关（P〈0．05）;Id-3蛋白表达在不同组织起源的胃癌之间差异有统计学意义（P〈0．05）;Claudin-1作为细胞间紧密连接的构成成份在胃癌组织中的表达作用与Id蛋白表达正相关。结论Claudin-1表达上调以及在胃癌侵袭过程中表达转化提示参与肿瘤的生物学行为的转化;Id-1在胃癌淋巴结转移的预测中有一定的应用价值;不同组织起源的胃癌Id-3呈差异性表达;Claudin及Id蛋白在胃癌的侵袭转移及临床治疗及预后判断有一定的应用价值。     

肾透明细胞癌组织中Cripto-1的表达及其对预后的预测价值

目的观察Cripto-1在肾透明细胞癌(ccRCC)中的表达情况,并统计分析其对预后的价值。方法收集76对ccRCC和周围正常肾组织的石蜡标本,应用免疫组织化学法(IHC)、RT-qPCR和Western blot检测其中Cripto-1的表达情况,并分析其表达与预后的关系。体外实验分析Cripto-1对细胞系的增殖、迁移、侵袭的影响。结果Cripto-1在ccRCC组织中的表达显著高于正常肾组织(P<0.05);Cripto-1高表达的患者预后更差;Cripto-1可以促进肿瘤细胞增殖,提高迁移和侵袭的能力。结论Cripto-1在肾透明细胞癌中表达升高,可以作为判断预后的一项指标,同时也可能成为治疗ccRCC的新靶点。     

PROX1通过上皮间质转化促进非小细胞肺癌进展

目的： 探讨转录因子Prospero同源框1（Prospero-related homeobox 1，PROX1）在非小细胞肺癌（non-small cell lung cancer，NSCLC）进展中的作用和机制。方法： 对含有108对非小细胞肺癌组织的组织芯片进行免疫组化染色，分析PROX1表达和NSCLC患者预后间的关系。采用Western印迹法和实时定量PCR分析PROX1在NSCLC细胞系中的表达情况和上皮间质转化的标志物。采用CCK-8、Transwell试验检测肿瘤细胞的增殖、迁移和侵袭能力。结果： PROX1在非小细胞肺癌组织中的表达上调。PROX1的表达与肿瘤直径、淋巴转移显著相关（P=0.003、0.042）。高表达PROX1的NSCLC患者其5年复发率高于PROX1低表达组（70.9%vs 50.9%，P=0.005），而5年生存率低于PROX1低表达组（49.1%vs 66.0%，P=0.016）。CCK-8和Transwell实验的结果显示，敲减PROX1后肿瘤的增殖、侵袭和转移能力明显减弱（P<0.01、P<0.05）。敲减PROX1后E-钙粘蛋白表达上调而波形蛋白表达下降（P<0.001）。结论： PROX1表达增高可通过促进上皮间质转化而促进NSCLC进展；PROX1是潜在的NSCLC预后预测因子。     

Immunohistochemical expression of MAP1LC3A and MAP1LC3B protein in breast carcinoma tissues

Autophagy is a protein degradation process within the cell and its deregulation has been linked to various diseases and the formation of cancer. One of the important proteins involved in the autophagy process is microtubule‐associated protein 1 light chain 3 (MAP1LC3). The aims of this study were to determine the MAP1LC3A and MAP1LC3B protein expression in both normal and cancer breast tissues and to determine the relationship between the expression of these proteins and type of tissues. Immunohistochemistry assessments were carried out on tissue microarrays consisting of breast tissues. MAP1LC3A expression was detected in 52/56 of normal breast tissue cores and 65/67 of breast cancer tissue cores. MAP1LC3B expression was detected in 55/56 of normal breast tissue cores and 67/67 of breast cancer tissue cores. MAP1LC3A and MAP1LC3B protein are expressed in the majority of normal and cancer breast tissues. A large number of MAP1LC3A and MAP1LC3B positive breast cancer tissues cores have high proportion of stained cells (81–100%) as compared with normal breast tissues. However, a significantly higher number of breast cancer tissues were found to express the MAP1LC3A protein with strong immunoreactivity as compared with the normal tissues, suggesting that MAP1LC3A may play a role in breast cancer development. J. Clin. Lab. Anal. 23:249–258, 2009. © 2009 Wiley‐Liss, Inc.     

甾醇类新药NSC67657诱导THP-1细胞分化及ICAT蛋白在细胞分化中的作用研究

目的 研究新的甲磺酸甾醇类药物NSC67657对白血病细胞的诱导分化作用及可能机制.方法 采用MTT法分析在不同浓度NSC67657作用下THP-1细胞的增殖水平,通过细胞表面分化抗原的检测,观察不同药物浓度、不同作用时间处理的THP-1细胞分化程度,并对完全分化细胞做形态学分析.通过RT-PCR和Western blot方法观察药物作用细胞前后β-catenin相关蛋白1(ICAT)基因和蛋白的表达情况;构建pDsRed-ICAT真核表达载体,测序后转染THP-1细胞,筛选阳性克隆,并做表达验证.采用流式细胞术,瑞特染色和超微结构观察,分析重组质粒转染细胞在药物处理前和处理24 h后THP-1细胞的分化情况.结果 通过比较可见药物处理后的THP-1细胞增殖明显受抑;细胞表面分化抗原CD14的表达水平随药物处理时间的延长和药物浓度的升高而增加,结合增殖分析,以10 μmol/L药物、连续诱导5 d为宜,细胞分化可达到90%以上.形态学观察验证了THP-1细胞在NSC67657的作用下向单核系分化.真核表达载体构建成功,电转后THP-1细胞ICAT基因和蛋白表达升高.药物作用前后,重组质粒转染THP-1细胞CD14的表达与对照组比较无明显差异;通过瑞特染色和超微结构观察,发现药物作用重组质粒转染THP-1细胞24 h后,细胞仍处初级分化阶段.结论 NSC67657可诱导THP-1细胞向单核系分化,并激活ICAT基因的表达,但仅是该基因的高表达并不 足以诱导THP-1细胞分化,也不会增加THP-1细胞对NSC67657 药物作用的敏感性.     

The tumor-suppressor gene LZTS1 suppresses hepatocellular carcinoma proliferation by impairing PI3K/Akt pathway

BackgroundTo study the role of LZTS1 in hepatocellular carcinoma (HCC) proliferation and the molecular mechanism involved.MethodsLZTS1 expression was studied in 10 HCC cell lines and 1 normal hepatocyte cell line by western blot analysis and qRT-PCR. One HCC cell line was selected and transfected with LZTS1 lentivirus. Cell proliferation and cell cycle were then determined by CCK-8 assay and flow cytometry, respectively. LZTS1, cyclin D1, CDK1, Cdc25C, pS473 Akt, and pT308 Akt mRNA and protein expressions were measured. PS473 Akt and pT308 Akt expression level was also compared with the HCC cells treated with LY294002.ResultsCompared with the normal hepatocyte cells, LZTS1 expression in HCC cells was significantly lower. After the transfection with LZTS1 lentivirus, HCC cell proliferation ability decreased markedly and HCC cells were blocked at G2/M phase. Cyclin D1 and CDK1 expression were both decreased but not significantly. Cdc25C expression was increased significantly. PS473 Akt and pT308 Akt expression level was increased significantly as well, which were almost the same with those transfected with LY294002.ConclusionLZTS1 could inhibit HCC cell proliferation by impairing PI3K/Akt pathway.     

核苷酸结合寡聚化结构域样受体蛋白3和caspase-1表达与非小细胞肺癌患者疾病进展的关系

目的 观察非小细胞肺癌(non-small cell lung cancer,NSCLC)患者外周血核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)及血清caspase-1 的表达情况,探...     

Upregulation of miR-370 contributes to the progression of gastric carcinoma via suppression of FOXO1

FOXO1 is downregulated in a number of cancers. However, the underlying mechanisms are poorly understood. In this study, we report that the expression of miR-370 was upregulated in gastric cancer cell lines and gastric cancer tissues. Overexpression of miR-370 in gastric cancer cells promoted the cell proliferation and anchorage-independent growth, while silencing of miR-370 showed opposite effects. miR-370-induced proliferation was correlated with the downregulation of cyclin-dependent kinase inhibitors, p27^Kip1 and p21^Cip1, and the upregulation of the cell cycle regulator cyclin D1. Furthermore, we identified that FOXO1 is the functional target of miR-370. Restored expression of FOXO1 together with miR-370 strongly abrogated miR-370-induced cell proliferation. Taken together, our results revealed a novel mechanism of FOXO1 suppression mediated by miR-370 in gastric cancer.     

HMGB1和MMP2在肾透明细胞癌中的表达及意义

目的研究高迁移率族蛋白B1（high mobility group box-1 protein,HMGB1）和基质金属蛋白酶2（matrix metalloproteinase-2,MMP2）在肾透明细胞癌（renal clear cell carcinoma,RCCC）中的表达及临床意义。方法收集58例RCCC、30例癌旁非癌肾组织、18例正常肾组织,采用免疫组织化学二步法检测HMGB1和MMP2的表达,并分析它们与临床病理参数之间的关系。58例RCCC中,组织学分级为高分化24例、中分化16例、低分化18例;临床分期为Ⅰ期13例、Ⅱ期15例、Ⅲ期21例、Ⅳ期9例。结果 HMGB1在正常肾组织、癌旁组织、RCCC中的阳性表达率分别为22.2%、36.6%、82.7%,呈现逐渐升高的趋势（P〈0.05）;在RCCC中,HMGB1蛋白阳性表达率在临床分期Ⅲ期和Ⅳ期高于Ⅰ期和Ⅱ期,有淋巴结转移组高于无淋巴结转移组,肿瘤直径≥5cm组高于肿瘤直径〈5cm组,肿瘤有坏死组高于无坏死组,HMGB1表达与年龄、性别、分化程度、肿瘤所在部位均无关（P〉0.05）。MMP2在正常肾组织、癌旁组织、RCCC中的阳性表达率分别为16.6%、33.3%、79.3%,呈现逐渐升高的趋势（P〈0.05）;在RCCC中,MMP2蛋白阳性表达率在临床分期Ⅲ期和Ⅳ期高于Ⅰ期和Ⅱ期,低分化组高于中、高分化组,有淋巴结转移组高于无淋巴结转移组,MMP2表达与年龄、性别、肿瘤所在部位、大小、是否坏死均无关（P〉0.05）。结论 HMGB1和MMP2蛋白的高表达参与了RCCC的发生、发展,可以作为判断RCCC预后的分子指标。     

Knockdown of Forkhead box A1 suppresses the tumorigenesis and progression of human colon cancer cells through regulating the phosphatase and tensin homolog/Akt pathway

BackgroundThis study aimed to evaluate the role and the underlying mechanisms of Forkhead box A1 (encoded by FOXA1) in colon cancer.MethodsWe analyzed FOXA1 mRNA and protein expression in colon cancer tissues and cell lines. We also silenced FOXA1 expression in HCT116 and SW480 cells to evaluate the effects on cell proliferation, cell cycle, migration, and invasion by using MTT, colony formation, flow cytometry, and the Transwell assay, respectively.ResultsFOXA1 immunostaining was higher in colon cancer tissues than adjacent healthy tissues. FOXA1 mRNA and protein expression was significantly increased in human colon cancer cells compared with a normal colonic cell line. FOXA1 expression was also significantly higher in colorectal cancer tissues from TCGA data sets and was associated with worse prognosis in the R2 database. FOXA1 expression was negatively correlated with the extent of its methylation, and its knockdown reduced proliferation, migration, and invasion, and induced G2/M phase arrest in HCT116 and SW480 cells by suppressing the phosphatase and tensin homolog/Akt signaling pathway and inhibiting epithelial–mesenchymal transition.ConclusionFOXA1 may act as an oncogene in colon cancer tumorigenesis and development.     

DKK-3和WIF-1基因启动子甲基化状态与肝细胞癌的关系研究

目的探讨DKK-3和WIF-1基因启动子甲基化状态与肝细胞癌发生的相关性。方法应用甲基化特异性聚合酶链反应检测33例肝细胞癌及相应的癌旁组织和20例正常对照肝组织中DKK-3和WIF-1基因启动子甲基化状态,分析上述组织中两种基因甲基化状态与临床资料的关系。结果DKK-3和WIF-1基因在癌组织中甲基化率明显高于癌旁组织和正常组织(P<0.05,P<0.05),而癌旁组织和正常组织中DKK-3和WIF-1基因甲基化率相比无明显差异;癌组织中DKK-3基因甲基化与临床资料中年龄及肝硬化有关;癌组织中WIF-1基因甲基化与临床资料中乙肝表面抗原、肝硬化有关。结论DKK-3和WIF-1基因启动子甲基化与HCC的发生相关;DKK-3和WIF-1基因致肝细胞癌的发生机制可能与肝硬化致肝细胞癌的发生机制不尽相同,两者是否有相互作用尚不清楚。