长链非编码 RNA MIAT 在肾透明细胞癌中的表达情况和预后意义

目的 探讨长链非编码RNAMIAT在肾透明细胞癌中的表达情况及与患者临床指标的相关性,分析其作为肾透明细胞癌分子标记物的可能性。方法 通过荧光定量PCR方法 检测MIAT在40例肾透明细胞癌组织和40例癌旁正常组织中的表达情况,同时结合TCGA数据库分析MIAT表达水平与肾透明细胞癌患者临床指标和预后的关系。结果 MIAT在肾透明细胞癌组织中的表达明显高于癌旁正常组织,在肾癌细胞系中的表达明显高于正常肾小管上皮细胞,差异均具有统计学意义（P<0.05）。TCGA数据库资料分析表明,MIAT表达水平与肾癌患者T分期（P<0.001）、M分期呈正相关（P<0.05）。Kaplan-Meier生存分析表明,高表达MIAT的肾癌患者总体生存时间明显低于低表达MIAT的肾癌患者（Log-rankP<0.05）。结论 MIAT在肾透明细胞癌组织和肾癌细胞系中高表达,有可能成为肾透明细胞癌的分子标记物。     

KCNN4 is a potential prognostic marker and critical factor affecting the immune status of the tumor microenvironment in kidney renal clear cell carcinoma

BackgroundThe tumor microenvironment (TME) has emerged as a crucial factor in cancer development and progression. Recent findings have indicated that tumor-infiltrating immune cells (TICs) in the TME may predict cancer prognosis and response to treatment. Herein, we sought to identify critical modulators of the kidney renal clear cell carcinoma (KIRC) TME.MethodsKIRC datasets from The Cancer Genome Atlas (TCGA) were analyzed using the ESTIMATE algorithm to determine the ImmuneScore and StromalScore. By profiling the differentially expressed genes (DEGs) in the ImmuneScore and StromalScore, we finally identified the immune- and stromal-related DEGs of the cases, through which we then performed intersection analysis to determine the immune-related genes (IRGs). Cox regression analysis and least absolute shrinkage and selection operator (LASSO) regression analysis were used to identify critical IRGs and construct a prognostic model. The CIBERSORT algorithm was used to calculate the relative content of 22 immune cell types. Finally, the datasets from the Gene Expression Omnibus (GEO) database were analyzed to validate results from the above analyses. Experimental validation was used on KIRC tissues by quantitative polymerase chain reaction (qPCR) and western blot.ResultsWe found that the ImmuneScore was negatively correlated with patients’ prognosis. Intersection analysis of the ImmuneScore and StromalScore identified 118 IRGs that were enriched in immune-related functions. Following IRGs screening by Cox and LASSO regression analyses, six genes were identified and used to construct a KIRC prognostic model. Intersection analysis of these six genes and protein-protein interaction (PPI) were performed and obtained the most critical gene: Potassium Calcium-Activated Channel Subfamily N Member 4 (KCNN4). Further analysis showed that KCNN4 expression was higher in tumor samples relative to normal controls, and was negatively correlated with prognosis. CIBERSORT analysis revealed significant correlation between KCNN4 expression and multiple types of TICs, demonstrating that KCNN4 may affect KIRC prognosis by influencing the TME immune status. Ultimately, the GEO datasets and validation experiments confirmed that KCNN4 was highly expressed in tumor tissues compared to the corresponding normal tissues.ConclusionsOur study demonstrated that KCNN4 might be a potential prognostic marker in KIRC, offering a novel therapeutic avenue.     

Identification of genes that correlate clear cell renal cell carcinoma and obesity and exhibit potential prognostic value

BackgroundRenal cell carcinoma (RCC) is a common urologic malignancy. Although the relationship between clear cell RCC (ccRCC) and obesity has been well-established by several large-scale retrospective studies, the molecular mechanisms and genetic characteristics behind this correlation remains unclear. In the current study, several bioinformatics tools were used to identify the key genes in ccRCC related to obesity.MethodsMicroarray data comparing ccRCC with normal renal tissues in patients with and without obesity were downloaded from the GEO database for screening of differentially expressed genes (DEGs). The DEGs were verified with expression level and survival analysis using several online bioinformatics tools.ResultsIn the current study, the differential expression of five genes correlated with both ccRCC and obesity; IGHA1 and IGKC as oncogenes, and MAOA, MUC20 and TRPM3 as tumor suppressor genes. These genes were verified by comparing the relationship between the expression levels and survival outcomes from open-source data in The Cancer Genome Atlas (TCGA) dataset.ConclusionsIn conclusion, the five genes differentially expressed in ccRCC and obesity are related to disease progression and prognosis, and therefore could provide prognostic value for patients with ccRCC.     

Expression and clinical significance of NUF2 in kidney renal clear cell carcinoma

BackgroundTo explore the expression and clinical significance of the cytokinesis-related gene NUF2 in kidney renal clear cell carcinoma (KIRC).MethodsGene expression profiles of KIRC patients were extracted from The Cancer Genome Atlas (TCGA) database. The differences in NUF2 mRNA expression between patients and controls, as well as the relationship between the clinical characteristics and overall survival of the patients, were analyzed. The expression of NUF2 protein in 83 cancer tissues and para-cancerous tissues was detected to analyze the relationship with clinical characteristics. Gene Set Enrichment Analysis (GSEA) was used to investigate the possible regulatory pathways of the NUF2 in the development of KIRC.ResultsNUF2 mRNA was significantly higher in patients with KIRC, and the prognosis of patients with high expression of NUF2 mRNA was significantly worse than those with low expression, and was related to the AJCC stage, T stage, lymph node metastases, and distant metastases. NUF2 mRNA was an independent prognostic risk factor for KIRC patients. The expression of NUF2 protein was significantly higher in KIRC patients than in paraneoplastic tissues and was markedly associated with the pathological grade. In addition, the high expression of NUF2 was associated with the upregulation of pathways such as homologous recombination and DNA replication.ConclusionsNUF2 may act as an independent prognostic biomarker for predicting the survival of KIRC patients.     

A preoperative clinical prognostic model for non-metastatic renal cell carcinoma

OBJECTIVE: To develop a model to predict the outcome before surgery for non-metastatic renal cell carcinoma (RCC). PATIENTS AND METHODS: The records of 660 patients with non-metastatic RCC, operated at three European medical institutes, were reviewed. Univariate and multivariate analyses were used to assess the clinical and pathological variables affecting disease-free survival. RESULTS: The median (range) follow-up was 42 (2-180) months; the disease recurred in 110 patients (16%). The 2- and 5-year overall survival was 87% and 54%, respectively. Five variables were significant in the univariate analysis, i.e. clinical presentation, clinical and pathological size, tumour grade and stage (P < 0.05). The preoperative variables, e.g. clinical presentation and clinical tumour size, were retained from the multivariate model. A recurrence risk formula (RRF) was constructed from this model, as (1.28 x presentation (asymptomatic = 0; symptomatic = 1) + (0.13 x clinical size)). Using this equation, the 2- and 5-year disease-free survival was 96% and 93% for an RRF of < or = 1.2 and 83% and 68% for an RRF of > 1.2. CONCLUSION: A formula was developed which, independent of stage, can be used to predict the rate of treatment failure in patients who undergo nephrectomy for non-metastatic RCC. The RRF might be useful for more accurate sub-grouping of good-prognosis patients, and for counselling patients before surgery, their personalized follow-up or adjuvant treatment once available.     

Assessment of the prognostic value of SPOCK1 in clear cell renal cell carcinoma: a bioinformatics analysis

BackgroundClear cell renal cell carcinoma (ccRCC) is one of the most common urological malignancies, and once metastasis occurs, it often has a poor prognosis and lacks effective treatment. Therefore, there is an urgent need to screen some new biomarkers and explore their molecular mechanisms to improve the early clinical diagnosis and targeted therapy of ccRCC. SPOCK1 (SPARC/osteonectin, CWCV and Kazal-like domains proteoglycan 1) is a conserved multi-domain proteoglycan that plays an important role in the development of multiple cancer types; however, its prognostic value in ccRCC has not been investigated. The study of the prognostic value of SPOCK1 in ccRCC is a good complement to the study of ccRCC biomarkers.MethodsDatabases of this study included Oncomine, Kaplan-Meier Plotter, GEPIA, GeneMANIA, cBioPortal, and TIMER. Student’s t-test was used to analyze the differences in SPOCK1 expression in ccRCC tissues compared with tumor-adjacent normal tissues. Kaplan-Meier curves for survival analysis were used to assess the correlation between the expression of SPOCK1 and the prognostic outcomes. Correlation module drew the expression scatterplots between SPOCK1 and immune cell infiltration in ccRCC, together with the Spearman’s rho value and estimated statistical significance.ResultsThe SPOCK1 mRNA expression was significantly higher in ccRCC tissues (mean expression ± SD: 920.2±195.2) than in normal tissues (mean expression ± SD: 358.4±29.1, P=0.008), and high SPOCK1 expression significantly and positively correlated with the pathological stage of ccRCC patients (F value =10.2, P<0.001). Higher expression of SPOCK1 was also associated with significantly shorter overall survival (OS) and disease-free survival (DFS) in ccRCC patients (GEPIA: P=0.046, P<0.001, respectively; Kaplan-Meier Plotter: P=0.002, P=0.0022, respectively). The function of SPOCK1 is mainly related to tumor development and extracellular matrix remodeling, and it may participate in the epithelial-mesenchymal transition process. SPOCK1 expression significantly and positively correlated with infiltration of several immune cells in ccRCC, including cancer-associated fibroblasts (CAFs) (Rho =0.333, P=2.16×10⁻¹³), tumor-associated macrophages (TAMs) (Rho =0.18, P=1.02×10⁻⁴), and tumor-associated neutrophils (TANs) (Rho =0.165, P=3.83×10⁻⁴). Conversely, there was a significant and negative correlation between SPOCK1 expression and infiltration of CD4⁺ T cells (Rho =−0.113, P=0.015).ConclusionsSPOCK1 may be a potential prognostic biomarker in ccRCC.     

基于TCGA数据库分析BIRC5在肾透明细胞癌中的表达及预后

目的通过分析杆状病毒凋亡抑制蛋白5(BIRC5)在肾透明细胞癌(ccRCC)组织中的表达,阐明BIRC5对其早期诊断及作为预后预测因子的作用。方法利用GEO、TCGA数据库和HPA分析BIRC5在ccRCC组织中mRNA和蛋白质水平的变化。运用UALCAN和LinkedOmics数据库阐述BIRC5表达与ccRCC临床病理学参数的相关性及对预后的影响。采用GEPIA、Kaplan-Meier Plotter、SurvExpress分析BIRC5表达与ccRCC预后的关系。结果BIRC5 mRNA在ccRCC中高表达,并且与ccRCC患者TNM分期相关(P<0.05),与ccRCC进展高度相关,高表达BIRC5mRNA是ccRCC患者不良预后指标。免疫组织化学结果证实,与正常肾组织相比,ccRCC中BIRC5蛋白表达量显著升高。结论在ccRCC中,BIRC5高表达是一种重要的早期诊断及不良预后指标,有望成为ccRCC早期诊断和预后预测标志物。     

Survivin和Vimentin在肾透明细胞癌中的表达及临床意义

目的探讨凋亡蛋白抑制因子生存素（Survivin）和波形蛋白（Vimentin）在肾透明细胞癌（clearcellrenalcellcarcinoma,ccRCC）中的表达及临床意义。方法采用免疫组化SP法检测56例ccRCC、25例癌旁组织及13例正常肾组织中Survivin和Vimentin的表达及定位情况。结果Survivin在肾透明细胞癌中高表达,而在癌旁及正常组织中未见表达;其表达与ccRCC临床分期及病理分级呈正相关,差异有统计学意义（P〈0．01）。Vimentin在正常肾组织及癌旁组织中亦未见表达,而在肾透明细胞癌中高表达,且与临床分期及病理分级呈正相关,差异有统计学意义（P〈0．01）。并且两个因子的表达呈显著正相关（r=0．566,P〈0．01）。结论Survivin和Vimentin在ccRCC组织中高表达,共同促进了ccRCC的发生、发展、侵袭与转移,联合两因子检测可为ccRCC的预后评估提供重要的信息。     

Identification of a novel immune-related microRNA prognostic model in clear cell renal cell carcinoma

BackgroundClear cell renal cell carcinoma (ccRCC) is a type of kidney cancer, and one of the most common malignant tumors. Many studies have shown that certain microRNAs (miRNAs) play an important role in the occurrence and development of ccRCC. Nevertheless, the prognosis of ccRCC patients is very rarely based on these “immuno-miRs”. Our aim was thus to determine the relationship between immune-related miRNA signatures and ccRCC.MethodsWe downloaded the miRNA expression data from 521 KIRC and 71 normal tissues in The Cancer Genome Atlas (TCGA). We used “limma” package and univariate Cox regression analysis to identify differentially expressed miRNAs (DEMs) that related to overall survival (OS). We applied lasso and multivariate Cox regression analyses to construct a prognostic model based on immuno-miRs. We evaluated the performance of model by using the Kaplan-Meier method. Furthermore, Cox regression analysis was used to determine independent prognostic signatures in ccRCC.ResultsA total of 59 significant immuno-miRs were identified. We use univariate Cox regression analysis to acquire 18 immune-related miRNAs which were markedly related to OS of ccRCC patients in the training set. We then constructed the 9-immune-related-miRNA prognostic model (miR-21, miR-342, miR-149, miR-130b, miR-223, miR-365a, miR-9-1, and miR-146b) by using lasso and multivariate Cox regression. Further analysis suggested that the immune-related prognostic model could be an independent prognostic indicator for patients with ccRCC. The prognostic performance of the 9-immune-related-miRNA prognostic model was further validated successfully in the testing set.ConclusionsWe established a novel immune-based prognostic model of ccRCC based on potential prognostic immune-related miRNAs. Our results indicated that the 9-miRNA signature could be a practical and reliable prognostic tool for ccRCC.     

Evaluation of the association of current cigarette smoking and outcome for patients with clear cell renal cell carcinoma

Objectives:   Cigarette smoking is a well known risk factor for the development of renal cell carcinoma (RCC); however, its association with tumor aggressiveness and patient outcome remains in question. Herein, we test the hypothesis that cigarette smoking is associated with a more aggressive phenotype and poorer outcome among patients with RCC.
Methods:   We examined data on 2242 patients treated with radical nephrectomy or nephron-sparing surgery for unilateral, sporadic, clear cell RCC at Mayo Clinic Rochester between 1970 and 2002. Associations of self-reported smoking status with death from RCC were assessed using Cox proportional hazards regression models summarized with hazard ratios (HR) and 95% confidence intervals (CI).
Results:   While former cigarette smoking was not associated with an increased risk of RCC death, current cigarette smokers were 31% more likely to die from RCC compared with non-smokers on a hazard ratio scale (HR 1.31; 95% CI 1.09–1.58; P  = 0.004). Interestingly, current smokers were more likely to present with advanced disease (i.e. later TNM stage) compared with both former and never smokers. After adjustment for TNM stage group and tumor grade, there was no longer a statistically significant increase in the risk of death from RCC for current cigarette smokers (HR 0.99; 95% CI 0.82–1.19; P  = 0.875).
Conclusions:   Patients who report current smoking at time of surgery are at increased risk of RCC death; however, this association is attenuated after adjustment for standard pathological indices and is therefore of little prognostic value. Nevertheless, the association of current smoking with more advanced disease at presentation (e.g. metastatic spread) warrants further investigation.     

C3, C3AR1, HLA-DRA,and HLA-E as potential prognostic biomarkers for renal clear cell carcinoma

BackgroundPrognostic biomarkers play a vital role in the early detection of the cancer and assessment of prognosis. With advances in technology, a large number of biomarkers of kidney renal clear cell carcinoma (KIRC) have been discovered, but their prognostic value has not been fully investigated, and thus have not been widely used in clinical practice. We aimed to identify the reliable markers associated with the prognosis of KIRC patients.MethodsWe obtained 72 normal samples and 539 tumor samples from The Cancer Genome Atlas (TCGA), and 23 normal samples and 32 tumor samples from the Gene Expression Omnibus (GEO). Overlapping differentially expressed genes (ODEGs) were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, followed by construction of a protein-protein interaction (PPI) network to screen hub genes. Kaplan-Meier analysis, univariate Cox analysis, multivariate Cox analysis, Wilcoxon signed-rank test, Kruskal-Wallis test, and gene set enrichment analysis (GSEA) were performed to verify the prognostic value and function of the markers we selected. The relationships among gene expression level, tumor immune cell infiltration, and immune-checkpoints were also analyzed.ResultsA total of 910 genes were screened out, and C3, C3AR1, HLA-DRA, and HLA-E were identified as potential tumor markers. The expression of each gene was closely associated with tumor immune cell infiltration, survival rate, and the patients’ clinical characteristics (P<0.05). C3AR1, HLA-DRA, and HLA-E were also verified as independent prognostic factors of KIRC (P<0.05), and all these potential biomarkers had a close correlation with immune checkpoints.ConclusionsC3, C3AR1, HLA-DRA, and HLA-E could be reliable biomarkers of KIRC and may have a significant contribution to make in immunotherapy, thus playing an important role in the improvement of prognosis.     

Validation of the World Health Organization/International Society of Urological Pathology grading for Chinese patients with clear cell renal cell carcinoma

BackgroundThis study aimed to compare the World Health Organization/International Society of Urological Pathology (WHO/ISUP) grading system and the Fuhrman grading system and to verify the WHO/ISUP grade as a prognostic parameter of clear cell renal cell carcinoma (ccRCC) in a Chinese population.MethodsThe study consisted of 753 ccRCC patients treated with curative surgery between 2010 and 2018 at Xiangya Hospital Central South University (Changsha, China). All pathologic data were retrospectively reviewed by two pathologists. Cancer-specific survival (CSS) and recurrence-free survival (RFS) were examined as clinical outcomes.ResultsAccording to the WHO/ISUP grading system (ISUP group), nephrectomy type, pT stage and WHO/ISUP grade were independent risk factors for CSS (P<0.0001, P=0.0127 and P<0.0001, respectively) and RFS (P<0.0001, P=0.0077, and P<0.0001, respectively). In the Fuhrman group, nephrectomy type, pT stage and Fuhrman grade were independent risk factors for CSS (P<0.0001, P=0.0004, and P<0.0001, respectively) and RFS (P<0.0001, P=0.0001, and P<0.0001, respectively). The C-index for CSS and RFS using the Fuhrman grading system was 0.6323 and 0.6342, respectively, and that using the WHO/ISUP grading system was 0.6983 and 0.7005, respectively, both higher than the former (P=0.0185, and P=0.0172, respectively). In addition, upgrading from Fuhrman grade 2 to ISUP grade 3 resulted in worse CSS and RFS for ccRCC patients (P=0.0033 and P =0.0003, respectively).ConclusionsWe first verified correlations between the postoperative prognosis and WHO/ISUP grade of ccRCC in a Chinese population and confirmed that the ability to predict clinical outcomes with the WHO/ISUP grading system was superior to that with the Fuhrman grading system.     

A postoperative prognostic nomogram predicting recurrence for patients with conventional clear cell renal cell carcinoma

PURPOSE: Few published studies have simultaneously analyzed multiple prognostic factors to predict recurrence after surgery for conventional clear cell renal cortical carcinomas. We developed and performed external validation of a postoperative nomogram for this purpose. We used a prospectively updated database of more than 1,400 patients treated at a single institution. MATERIALS AND METHODS: From January 1989 to August 2002, 833 nephrectomies (partial and radical) for renal cell carcinoma of conventional clear cell histology performed at Memorial Sloan-Kettering Cancer Center were reviewed from the center's kidney database. Patients with von Hippel-Lindau disease or familial syndromes, as well as patients presenting with synchronous bilateral renal masses, or distant metastases or metastatic regional lymph nodes before or at surgery were excluded from study. We modeled clinicopathological data and disease followup for 701 patients with conventional clear cell renal cell carcinoma. Prognostic variables for the nomogram included pathological stage, Fuhrman grade, tumor size, necrosis, vascular invasion and clinical presentation (ie incidental asymptomatic, locally symptomatic or systemically symptomatic). RESULTS: Disease recurrence was noted in 72 of 701 patients. Those patients without evidence of disease had a median and maximum followup of 32 and 120 months, respectively. The 5-year probability of freedom from recurrence for the patient cohort was 80.9% (95% confidence interval 75.7% to 85.1%). A nomogram was designed based on a Cox proportional hazards regression model. Following external validation predictions by the nomogram appeared accurate and discriminating, and the concordance index was 0.82. CONCLUSIONS: A nomogram has been developed that can be used to predict the 5-year probability of freedom from recurrence for patients with conventional clear cell renal cell carcinoma. This nomogram may be useful for patient counseling, clinical trial design and effective patient followup strategies.     

Development and validation of the prognostic value of the immune-related genes in clear cell renal cell carcinoma

BackgroundClear cell renal cell carcinoma (ccRCC) is a highly heterogeneous tumor, resulting a challenge of developing target therapeutics. Not long ago, immune checkpoint blockade regimens combine with tyrosin kinase inhibitors have evolved frontline options in metastatic RCC, which implies arrival of the era of tumor immunotherapy. Studies have demonstrated immune-related genes (IRGs) could characterize tumor milieu and related to patient survival. Nevertheless, the clinical significance of classifier depending on IRGs in ccRCC has not been well established.MethodsThe R package limma, univariate and LASSO cox regression analysis were used to screen the prognostic related IRGs from TCGA database. Multivariate cox regression was utilized to establish a risk prediction model for candidate genes. Quantitative real-time PCR was used to confirm the expression of candidates in clinical samples from our institution. CIBERSORT algorithm and correlation analysis were applied to explore tumor-infiltrating immune cells signature between different risk groups. A clinical nomogram was also developed to predict OS by using the rms R package based on the risk prediction model and other independent risk factors. The ICGC data was used for external validation of either gene risk model or nomogram.ResultsWe identified 382 differentially expressed immune related genes. Four unique prognostic IRGs (CRABP2, LTB4R, PTGER1 and TEK) were finally affirmed to associate with tumor survival independently and utilized to establish the risk score model. All candidates’ expression was successfully laboratory confirmed by q-PCR. CIBERSORT analysis implied patients in unfavorable-risk group with high CD8 T cell, regulatory T cell and NK cell infiltration, as well as high expression of PD-1, CTLA4, TNFRSF9, TIGIT and LAG3. A nomogram combined IRGs risk score with age, gender, TNM stage, Fuhrman grade, necrosis was further generated to predict of 3- and 5-year OS, which exhibited superior discriminative power (AUCs were 0.811 and 0.795).ConclusionsOur study established and validated a survival prognostic model system based on 4 unique immune related genes in ccRCC, which expands knowledge in tumor immune status and provide a potent prediction tool in future.     

术前中性粒细胞与淋巴细胞比值和血小板与淋巴细胞比值对肾透明细胞癌的预后评估价值

目的:探究术前外周血中性粒细胞与淋巴细胞比值(NLR)和血小板与淋巴细胞比值(PLR)在评估肾透明细胞癌(ccRCC)患者预后的作用。方法:回顾性分析2001年12月—2010年12月在我院接受手术治疗的352例肾细胞癌(RCC)患者的临床资料,年龄25~82岁,平均(55.1±12.2)岁;随访时间1~200个月,平均(106.1±35.1)个月;中位总生存期(OS)为104个月,中位无复发生存期(RFS)为101个月。通过受试者工作特征曲线(ROC)确定NLR及PLR的最佳临界值并进行分组,通过Kaplan-Meier法和Cox回归对RCC患者中的NLR及PLR进行预后分析。结果:按最佳临界值NLR<2.05(155例)及≥2.05(197例)、PLR<140(236例)及PLR≥140(116例)对患者进行分组。高NLR及PLR与大肿瘤直径(P=0.026,P=0.019)、高肿瘤TNM分期(P=0.003,P<0.001)、高肿瘤Fuhrman分级(P=0.021,P=0.008)及转移或复发有关(P<0.001,P<0.001)。相比于单独使用NLR或PLR,联合NLR及PLR能够更有效地预测OS及RFS。Cox多因素分析结果提示高NLR(P<0.001)、高PLR(P=0.004)、患者年龄≥60岁(P<0.001)、大肿瘤直径(P=0.043)、高肿瘤TNM分期(P<0.001)、高肿瘤Fuhrman分级(P<0.001)与患者OS相关,并且高NLR(P=0.012)、高PLR(P=0.014)、高肿瘤TNM分期(P<0.001)、高肿瘤Fuhrman分级(P=0.002)与患者RFS相关。结论:术前NLR及PLR是ccRCC患者术后OS及RFS的独立预后因素。高NLR、PLR预示着ccRCC患者较高的复发转移风险及较差的生存预后。