Downregulation of long non-coding RNA ENSG00000241684 is associated with poor prognosis in advanced clear cell renal cell carcinoma

Objective

In order to identify potential novel biomarkers of advanced clear cell renal cell carcinoma (ccRCC), we re-evaluated published long non-coding RNA (lncRNA) expression profiling data.

基于生物信息学分析铁死亡相关lncRNA对肾透明细胞癌组织免疫浸润和患者预后评估的意义

目的：采用生物信息学方法探索与肾透明细胞癌(ccRCC)组织中铁死亡相关的lncRNA,并探讨其与免疫细胞浸润及患者预后的相关性,为ccRCC患者提供新的分子靶点。方法：从癌症基因组图谱(TCGA)数据库下载cc RCC的转录本数据和临床数据,利用单样本基因集富集分析(ssGSEA)及相关性分析获得与铁死亡相关的lncRNA;通过单因素和多因素回归分析构建与铁死亡相关的lncRNA特征图,分析其与预后的关系;利用R软件分析铁死亡相关lncRNA与肿瘤免疫细胞浸润和药物敏感性之间的关系。构建铁死亡相关RNA网络,并通过qPCR验证中国人ccRCC组织和癌旁组织（取自2019年12月至2021年03月间在西南医科大学附属医院手术切除8例标本）中关键lncRNA的表达。结果：Kaplan-Meier分析表明,铁死亡评分高的患者总OS率低于铁死亡评分低的患者。单因素和多因素回归分析确定11个ccRCC铁死亡相关lncRNA可评估患者预后,并构建ccRCC患者1、3、5年预后预测列线图。免疫细胞浸润分析表明,铁死亡相关lncRNA与ccRCC免疫细胞浸润密切相关,其中LINC01871、PRKA...     

lncRNA NONHSAT113026在肾透明细胞癌中的表达及功能

目的:本研究旨在探讨lncRNA NONHSAT113026（lncRNA 026）在肾透明细胞癌（clear cell renal cell carcinoma,ccRCC）组织中的表达及对肾癌细胞增殖、迁移和侵袭的影响。方法:采用荧光定量聚合酶链反应（quantitative real-time polymerase chain reaction,qRT-PCR）分别在83例肾透明细胞癌和对应癌旁组织、肾癌细胞株786-O及ACHN中检测lncRNA 026的表达水平,并分析其表达水平与患者临床病理参数的相关性。通过慢病毒载体技术构建稳定过表达lncRNA 026的肾癌细胞786-O及ACHN,采用细胞计数试剂盒（cell counting kit-8,CCK-8）检测细胞增殖活性,Transwell小室检测细胞转移能力,蛋白印迹法（Western blot）分析PI3K/Akt信号通路中相关蛋白的表达情况。结果:与癌旁组织相比,lncRNA 026在ccRCC组织中的表达水平显著下调（P＜0.000 1）;在肾癌细胞786-O和ACHN中过表达lncRNA 026能显著抑制细胞的增殖、迁移和侵袭（P＜0.01）;Western blot结果显示,过表达lncRNA 026可下调PI3K/Akt信号通路中p-PI3K、p-Akt、GSK-3β的表达（P＜0.01）。结论:lncRNA 026在肾透明细胞癌组织中低表达,并且过表达lncRNA 026能显著降低肾癌细胞的增殖、侵袭和迁移能力,其作用机制可能与其抑制PI3K/Akt信号通路活化有关。提示lncRNA 026在肾癌中发挥了重要作用,有望成为治疗肾癌的潜在药物作用靶点。     

Piwi-interacting RNAs as novel prognostic markers in clear cell renal cell carcinomas

Background

Piwi-interacting RNAs (piRNAs) are small RNAs of 27–30 nucleotides mapping to transposons or clustering in repeat genomic regions. Preliminary studies suggest an important role in cancerogenesis. This study is the first one investigating their prognostic impact in clear cell renal cell cancer (ccRCC) patients.

Methods

Three piRNAs (piR-30924, piR-57125, and piR-38756) selected on the basis of initial piRNA microarray analyses were determined using RT-qPCR in non-metastatic (n = 76) and metastatic (n = 30) ccRCC tissue at the time of nephrectomy in comparison to normal renal tissue (n = 77) and tissue from distant ccRCC metastases (n = 13). Primary clinical end points were recurrence-free and overall survival.

Results

piR-57125 showed lower expression in metastatic than in non-metastatic tumors, whereas the expression of piR-30924 and piR-38756 increased in metastatic tumors. The higher expression of piR-30924 and piR-38756 as well as the lower expression of piR-57125 in metastatic primary tumors were significantly associated with tumor recurrence and overall survival. Multivariate Cox regression analyses revealed both piR-30924 and piR-57125 as independent prognostic predictors. This impact was even more pronounced in non-metastatic patients.

Conclusions

This study demonstrates that the expression levels of these piRNAs in primary non-metastatic and metastatic ccRCC tissue can serve as potential prognostic biomarkers in combination with clinicopathological factors.

Electronic supplementary material

The online version of this article (doi:10.1186/s13046-015-0180-3) contains supplementary material, which is available to authorized users.     

术前血清白球比预测局限性和局部进展性肾透明细胞癌患者预后的单中心回顾性研究

目的:评估术前白蛋白球蛋白比值,即白球比（AGR）与局限性及局部进展性肾透明细胞癌（ccRCC）患者临床和病理特征的相关性,评估其预后价值。方法:回顾性收集苏州大学附属第三医院415例在2003年至2012年间经肾切除手术局限性和局部进展性ccRCC患者的临床病理数据及随访资料,整合系统炎性指标及传统预后评估系统,绘制ROC曲线。结果:AGR越小,肿瘤的恶性程度越高;AGR是研究对象的独立预后因素（HR:6.799,95%CI:3.215~14.377,P＜0.01）;与现有的系统炎性指标与传统预后评估系统相比,AGR对局限性及局部进展性ccRCC的预后预测能力更强。结论:AGR是一个预测局限性和局部进展性ccRCC患者术后总体生存期（OS）的可靠指标,是患者OS独立的危险因子,相较系统炎性指标与传统预后评估系统有更高的实用价值。     

TNFAIP3在透明细胞肾细胞癌中的表达及临床意义

目的:探讨TNFAIP3在透明细胞肾细胞癌（clear cell renal cell carcinoma,ccRCC）中的表达情况,并分析其表达与患者临床病理特征的相关性。方法:利用GEPIA2数据库分析TNFAIP3在ccRCC组织和正常肾脏组织中的差异表达,收集具备完整临床资料的97例ccRCC组织及对应的癌旁正常肾脏组织,采用免疫组织化学EnVision两步法检测TNFAIP3蛋白在ccRCC组织及对应癌旁正常肾脏组织中的表达,分析TNFAIP3蛋白的表达情况与患者年龄、性别、WHO/ISUP分级、肿瘤最大径及TNM分期等临床病理特征的关系。结果:GEPIA2数据库分析显示,与正常肾脏组织相比TNFAIP3在ccRCC组织中呈高表达（P＜0.05）,但TNFAIP3高表达ccRCC患者总生存时间和无瘤生存时间均显著高于TNFAIP3低表达ccRCC患者（P＜0.05）;TNFAIP3免疫组化检测证实,TNFAIP3在ccRCC组织中呈淡黄至棕褐色表达于细胞膜和细胞质,在ccRCC组织中高表达比例明显高于癌旁正常肾脏组织（P＜0.05）,TNFAIP3表达水平与ccRCC组织的WHO/ISUP肾细胞癌分级、肿瘤最大径、TNM分期、有无淋巴结转移临床参数有关（P＜0.05）,TNFAIP3高表达则提示较好的预后。结论:ccRCC组织中TNFAIP3的表达明显升高,而TNFAIP3高表达的ccRCC患者总生存期和无瘤生存期均好于TNFAIP3低表达的ccRCC患者,TNFAIP3参与ccRCC增殖、转移中的作用方式与分子调控机制则待进一步研究。     

宫颈鳞状细胞癌组织LncRNA EXOC7表达与预后相关性研究

目的 宫颈癌是女性生殖系统常见的恶性肿瘤之一.长链非编码RNA(lncRNA)的异常表达与肿瘤的发生、发展密切相关.本研究拟探讨长链非编码RNA外泌体复合物7(Long non-coding RNA Exocyst complex component 7,LncRNA EXOC7)在宫颈鳞状细胞癌(cervical squamous cell cancer,SCC)组织中的表达及临床意义.方法 收集2012-01-01-2016-12-30四川省人民医院肿瘤科及妇产科行手术及穿刺活检的98例SCC患者组织标本,其中癌旁组织(距癌组织＞2 cm)58例;同期收集该院40例正常宫颈组织.采用QRT-PCR法检测SCC组织及癌旁组织标本中Ln-cRNA EXOC7的表达,采用单因素及多因素Cox回归分析LncRNA EXOC7的表达与患者临床病理特征及预后的关系,生存曲线分析LncRNA EXOC7与患者总生存时间(overall survival,OS)及无进展生存时间(progression free survival,PFS)的关系.结果 SCC组织中lncRNA EXOC7的平均表达量为(6.805±1.475),明显高于癌旁组织(1.510±0.860)及正常宫颈组织(1.476±0.33)的平均表达量,差异有统计学意义,F=74.43,P＜0.001.LncRNA EXOC7高表达与患者的FIGO分期、淋巴结转移和肿瘤浸润深度相关,差异均有统计学意义,均P＜0.05;高表达组LncRNA EXOC7患者PFS为(12.00±1.55)个月,较低表达组(27.35±2.25)个月明显缩短,差异有统计学意义,x2=51.650,P＜0.001;高表达组LncRNA EXOC7患者OS为(25.08±3.32)个月,较低表达组(44.80±2.92)个月明显缩短,差异有统计学意义,x2=38.23,P＜0.001;Cox多因素分析显示,LncRNA EXOC7的表达是SCC患者的独立预后因素,HR=3.750,95％CI为1.530～7.97,P＜0.001.结论 高表达LncRNA EXOC7 SCC患者的预后差,LncRNA EXOC7可能作为潜在的SCC治疗靶点及新预后评估分子标志物.     

Integrated analysis of long noncoding RNA associated‐competing endogenous RNA as prognostic biomarkers in clear cell renal carcinoma

Clear cell renal cell carcinoma (ccRCC) is one of the most common malignant carcinomas and its molecular mechanisms remain unclear. Long noncoding RNA (lncRNA) could bind sites of miRNA which affect the expression of mRNA according to the competing endogenous (ceRNA) theory. The aim of the present study was to construct a ceRNA network and to identify key lncRNA to predict survival prognosis. We identified differentially expressed mRNA, lncRNA and miRNA between tumor tissues and normal tissues from The Cancer Genome Atlas database. Then, using bioinformatics tools, we explored the connection of 89 lncRNA, 10 miRNA and 22 mRNA, and we constructed the ceRNA network. Furthermore, we analyzed the functions and pathways of 22 differentially expressed mRNA. Then, univariate and multivariate Cox regression analyses of these 89 lncRNA and overall survival were explored. Nine lncRNA were finally screened out in the training group. The patients were divided into high‐risk and low‐risk groups according to the 9 lncRNA and low‐risk scores having better clinical overall survival (P < .01). Furthermore, the receiver operating characteristic curve demonstrates the predicted role of the 9 lncRNA. The 9‐lncRNA signature was successfully proved in the testing group and the entire group. Finally, multivariate Cox regression analysis and stratification analysis further proved that the 9‐lncRNA signature was an independent factor to predict survival. In summary, the present study provides a deeper understanding of the lncRNA‐related ceRNA network in ccRCC and suggests that the 9‐lncRNA signature could serve as an independent biomarker to predict survival in ccRCC patients.     

小细胞肺癌组织中lncRNA表达谱的差异

目的：观察长链非编码RNA（ lncRNA）在未发生转移的小细胞肺癌和发生转移的小细胞肺癌组织中表达的差异。方法：利用高通量lncRNA芯片技术分别检测3例未发生转移的小细胞肺癌组织和3例发生转移的小细胞肺癌组织中lncRNA表达谱的变异,经对原始数据进行预处理达到均一化后,筛选出差异表达的lncRNA,进行聚类分析。结果：将lncRNA表达在发生转移的小细胞肺癌组织中与未发生转移的小细胞肺癌组织中的变化倍数在2倍以上并有显著差异（ P＜0.05）的lncRNA,确定为差异性表达的lncRNA。其中在3例发生远处转移的患者肺癌组织中变化均在2倍以上的共842条,占所有lncRNA的12.1％。其中,2倍以上表达上调的共440条,2倍以上表达降低的共402条;5倍以上表达升高的共31条;5倍以上表达降低的共65条。结论：发生转移的小细胞肺癌组织与未发生转移的小细胞肺癌组织比较,lncRNA表达谱发生显著变化。提示差异性表达的lncRNAs参与了小细胞肺癌侵袭和转移的恶性表型。     

沉默lncRNA TUG1对肾透明细胞癌细胞增殖、凋亡及迁移的影响

目的:探究沉默lncRNA TUG1对肾透明细胞癌（clear cell renal cell carcinoma,ccRCC)细胞增殖、凋亡及迁移的影响。方法:选用37例ccRCC组织及癌旁组织样本,通过提取样本总RNA并行逆转录及转染,利用qRT-PCR检测lncRNA TUG1在ccRCC组织、细胞系及癌旁组织中的表达,并通过CCK-8法、流式细胞检测法及划痕实验检测沉默lncRNA TUG1对ccRCC细胞A704、A498增殖、凋亡及迁移的影响,采用统计软件进行数据处理,探究lncRNA TUG1在ccRCC中的表达及临床意义。结果:qRT-PCR检测lncRNA TUG1表达情况,结果显示:lncRNA TUG1在ccRCC组织中的表达显著高于癌旁组织,在肾细胞癌细胞系A704和A498中的表达明显高于正常细胞系HK2,差异有统计学意义（P＜0.05）;CCK-8检测细胞增殖情况,结果显示:A704和A498细胞经转染干扰后细胞增殖活性较对照组显著降低（P＜0.05）,沉默lncRNA TUG1可抑制细胞增殖;流式细胞仪检测细胞凋亡情况,结果显示:A704和A498细胞凋亡比例较空白对照组明显增多（P＜0.05）,沉默lncRNA TUG1可促进细胞凋亡;划痕实验检测细胞迁移能力,结果显示:A704和A498细胞迁移能力较空白对照组降低（P＜0.05）,沉默lncRNA TUG1可抑制细胞迁移。结论:沉默lncRNA TUG1可诱导ccRCC细胞凋亡,抑制其增殖、迁移,可作为临床研究的新靶点。     

长链非编码RNA MAFG-AS1在骨肉瘤中的表达及功能

目的:探讨及分析骨肉瘤中差异表达长链非编码RNA（long non-coding RNA,lncRNA）的表达谱及功能。方法:本课题前期通过5对组织（骨肉瘤组织和瘤旁组织）进行高通量转录组学测序筛选出差异表达的lncRNAs,并筛选出上调变化的前5位lncRNAs,通过荧光定量PCR在骨肉瘤细胞系内检测lncRNA MAFG-AS1的差异表达,并通过小干扰RNA技术干扰其表达,CCK-8法检测敲低lncRNA后细胞增殖是否发生变化,Western-blotting检测下游蛋白变化。结果:高通量数据分词浅析显示,共有376个lncRNAs在骨肉瘤组织中差异表达,其中206个上调,170个下调,并筛选出上调变化的前5位lncRNAs。荧光定量PCR显示lncRNA MAFG-AS1在骨肉瘤细胞内高表达。CCK-8法结果显示该lncRNA MAFG-AS1具有影响骨肉瘤细胞增殖的作用,且下游蛋白RRM2表达发生变化。结论:骨肉瘤组织中存在明显的lncRNAs差异性表达。且lncRNA MAFG-AS1在骨肉瘤发生和发展中发挥中重要的作用。     

Epithelial cell adhesion molecule is an independent prognostic marker in clear cell renal carcinoma

Epithelial cell adhesion molecule (EPCAM) has recently attained a renewed interest as a candidate protein in diagnosis, prognostication and therapy of various tumor entities. The molecular epidemiology and prognostic relevance of EPCAM in renal cell carcinoma (RCC) and amongst the histological subtypes of RCC are unclear. We analyzed the prevalence and prognostic significance of EPCAM in a tumor tissue microarray composed of 1,088 independent RCCs samples by immunohistochemistry (IHC). We found significant variations of EPCAM IHC staining intensities in between the RCC subtypes: in papillary and chromophobe RCC, the majority of tumors (89–93%) showed an at least weak EPCAM protein expression. In the largest subgroup, the clear cell (cc)RCC (n = 767), a negative EPCAM IHC was found in 1/3 of the patients and was associated with high‐grade disease and nodal metastases. Kaplan–Meier analyses demonstrated a significant association between positive EPCAM IHC and prolonged overall survival, even in a subset of low‐risk ccRCC. In multivariable analyses, EPCAM represented an independent risk factor of survival throughout all subgroups. For localized, low‐grade ccRCC, information of EPCAM IHC raised predictive accuracy of a multivariate model by ～5%, compared to T‐stage and grade alone. Our findings indicate that EPCAM is an independent prognostic molecular marker in ccRCC and, especially in localized ccRCC, might be able to provide auxiliary information for a better prognostication.     

Lymphangiogenesis in kidney cancer: expression of VEGF-C, VEGF-D and VEGFR-3 in clear cell and papillary renal cell carcinoma

The vascular endothelial growth factors VEGF-C, VEGF-D and its receptor, VEGFR-3, are overexpressed in different malignancies and associated with lymph node metastasis and poor prognosis. We analysed these factors in clear cell (ccRCC) and papillary (pRCC) renal cell carcinoma (RCC). The results were correlated with various clinicopathological parameters (CPP). We constructed a tissue microarray with tumor samples of 135 (81%) ccRCC and 31 (19%) pRCC. After immunohistochemical staining using polyclonal antibodies for VEGF-C, VEGF-D and VEGFR-3, a semiquantitative analysis was performed to determine the levels of expression. The results were compared between the two subgroups and were correlated with CPP. In the two subgroups the expression of VEGF-C was significantly correlated with that of VEGF-D (p<0.001). There was an increased expression of VEGF-C in 11% of ccRCC and 36% of pRCC (p=0.002). VEGF-D expression was positive by means of analysis in 22% of ccRCC and 42% of pRCC (p=0.039). There was no significant difference regarding the expression of VEGFR-3 between the subgroups (44% ccRCC and 61% pRCC, p=0.11). No correlation was found between the expression of the analysed parameters and CPP (TNM, grading, progression-free survival and overall survival) in either the entire group or in the two subgroups. In summary, ccRCC and pRCC show a different expression pattern of the analysed lymphangiogenic factors. Further studies are necessary to confirm these results and to determine whether the VEGF-C/VEGF-D/VEGFR-3-axis can play a role as a prognostic tool or a target for therapeutic intervention in renal cell carcinoma.     

High expression levels of survivin protein independently predict a poor outcome for patients who undergo surgery for clear cell renal cell carcinoma

BACKGROUND: In a previous study of gene array data, the authors identified survivin as a candidate marker of aggressiveness in clear cell renal cell carcinoma (ccRCC). What remained in question was whether survivin expression at the protein level is an independent predictor of disease progression and cancer-specific survival. METHODS: Between 1990 and 1994, 312 patients underwent nephrectomy for ccRCC at Mayo Clinic Rochester and had paraffin tissue available. The authors performed immunohistochemistry with antisurvivin antibody, quantitated the expression by using an image-analysis system, and analyzed the association of survivin expression with disease progression and cancer-specific survival. RESULTS: Within the cohort, 97 patients (31.1%) had high levels of survivin expression. Patients who had high survivin expression levels were at significantly increased risk of death from RCC compared with patients who had low expression levels (risk ratio [RR], 5.3; 95% confidence interval [95% CI], 3.5-7.9). The 5-year cancer-specific survival rate was 43.0% for patients with high survivin expression and 87.2% for patients with low survivin expression. In multivariate analysis, survivin expression remained associated with death from RCC even after adjusting for the Eastern Cooperative Oncology Group performance status; 2002 Tumor, Lymph Node, Metastases (TNM) stage groupings and nuclear grade (RR, 2.4; 95%CI, 1.5-3.8); and the Mayo Clinic composite TNM stage groupings, tumor size, nuclear grade, and tumor necrosis (SSIGN) score (RR, 1.8; 95%CI, 1.1-2.9). Among 273 patients who had localized ccRCC, survivin expression was associated significantly with cancer progression (RR, 3.9; 95%CI, 2.4-6.2). CONCLUSIONS: Survivin expression is an independent predictor of ccRCC progression and death from RCC. Thus, survivin has the potential to offer additional prognostic information and to provide a novel target for the development of new adjuvant therapies.     

Survivin and HLA-I expression predicts survival of patients with clear cell renal cell carcinoma

Altered expression of survivin and leukocyte antigen class I (HLA-I) proteins is associated with tumor progression. This study investigated their expressions in clear cell renal cell carcinoma (ccRCC) tissues for association with a clinical significance of ccRCC patients. Ninety ccRCC and 20 normal tissue samples (i.e., control) were immunohistochemically stained for survivin and HLA-I expression for an association with clinicopathological data and survival of ccRCC patients. Survivin protein was expressed in 82.2 % (74/90) of ccRCC tissue samples compared to 0 % in the normal tissues, and HLA-I protein was expressed in 90 % (18/20) of the normal tissues vs. 67.8 % (61/90) in ccRCC samples. Survivin expression was associated with tumor grade, stage, and lymph node metastasis (p?=?0.000, p?=?0.016, and p?=?0.001, respectively). Conversely, lost HLA-I expression did not have any associations with clinicopathological data (p?>?0.05). Survivin-negative patients had a higher tumor-free survival rate than patients with survivin expression (p?=?0.037). Patients with normal HLA-I levels had a higher tumor-free survival rate than those with reduced HLA-I levels (p?=?0.02). The uni- and multivariate analyses indicated that expression of survivin and HLA-I, individually and in combination, was an independent predictor for survival of ccRCC patients. Overexpression of survivin but reduced HLA-I expression is useful in the prediction of tumor-free survival of ccRCC patients.