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.     

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.     

胰腺转移性肾透明细胞癌的临床病理特征和预后分析

目的探讨胰腺转移性肾透明细胞癌患者的临床病理特征和预后。方法回顾性分析2000年1月至2018年12月在北京协和医院收治的经病理确诊的18例胰腺转移性肾透明细胞癌患者的临床病理资料。结果18例患者中,男11例,女7例;确诊肾透明细胞癌的平均年龄为51.4岁。其中左肾8例(44.4%),右肾10例(55.6%);同时性转移3例,异时性转移15例,从确诊肾透明细胞癌到发现胰腺转移的中位发生转移的时间为156个月,主要临床表现有腹痛、黄疸、消化道出血、恶心、乏力、体重下降等。其中胰腺单病灶患者7例(38.9%),多病灶患者11例(66.1%),9例患者(50.0%)除胰腺外还同时存在其他部位转移。5例患者行胰腺转移灶切除,15例患者服用靶向药物治疗。随访时间1~361.5个月,平均随访时间为171.7个月,死亡5例,生存13例,中位生存时间为122个月,5年生存率为81.4%。是否为同时性转移、是否为10年后复发、纪念斯隆-凯特琳癌症中心模型预后评分以及国际转移性肾细胞癌联合数据库评分是影响胰腺转移性肾透明细胞癌患者预后的因素。结论胰腺转移性肾透明细胞癌罕见,但预后较好,尤其是10年后复发转移至胰腺的患者,行手术切除胰腺转移灶未发现明显的生存获益。     

MYC pathway is activated in clear cell renal cell carcinoma and essential for proliferation of clear cell renal cell carcinoma cells

Clear cell renal cell carcinoma (ccRCC) is the major and aggressive subtype of RCC. Previously, we identified 383 differentially expressed genes by analyzing full-length cDNA libraries of ccRCC and normal kidney tissues. In this study, we applied functional network analysis to the differentially expressed genes for identifying deregulated molecular pathways in ccRCC, and the results indicated that MYC showed a prominent role in the highest scoring network. The upregulation of MYC expression was validated in ccRCC tissues and cell lines. Furthermore, Knockdown of MYC expression by MYC-specific siRNA significantly inhibited the abilities of uncontrolled proliferation, anchorage-independent growth and arrested cell cycle in the G0/G1 phase in ccRCC cells. Moreover, we found that 37 differentially expressed genes were shown to be MYC-target genes, and the upregulation of the MYC-target genes BCL2, CCND1, PCNA, PGK1, and VEGFA were demonstrated. The expression of these MYC-target genes was significantly correlated with the expression of MYC in ccRCC tissues, and knockdown of MYC also suppressed the expression of these MYC-target genes in ccRCC cells. The recruitment of MYC to the promoter regions of BCL2, CCND1, PCNA, PGK1, and VEGFA was shown by Chromatin immunoprecipitation assay. These results suggest that MYC pathway is activated and plays an essential role in the proliferation of ccRCC cells.     

53例国人肾透明细胞癌中PBRM 1基因的突变分析

目的 分析Polybromo 1（PBRM 1）基因在国人肾透明细胞癌中的突变特点,探讨基因突变与临床病理特征和预后的关系。方法 选取53例肾透明细胞癌患者,利用PCR结合直接测序的方法检测PBRM 1基因2～5号外显子、15～17号外显子的突变情况。分析PBRM 1基因突变与临床病理特征和预后的相关性。结果 53例样本中检测出PBRM 1基因突变14例,突变率为26％。突变较集中分布在第15、17外显子,第15外显子5例,第17外显子6例。PBRM 1基因突变组和无突变组的中位无进展生存期分别为5个月（95％CI：1.6～8.4个月）和14个月（95％CI：9.7～18.3个月）,中位生存期分别为7个月（95％CI：2.6～11.4个月）和18个月（95％CI：5.6～30.4个月）,差异均有统计学意义（P＜0.05）。 PBRM 1基因突变与性别、年龄、发病部位、分期及疾病控制率等无关（P＞0.05）。结论 中国人肾透明细胞癌中PBRM 1基因突变率较高,突变主要位于第15、17外显子。PBRM 1基因突变可能是肾透明细胞癌患者预后不良的危险因素。     

RhoB在肾透明细胞癌中的表达及临床意义

目的:探讨RhoB在肾透明细胞癌中的表达及临床意义.方法:采用实时定量PCR、Western blot和免疫组化方法检测RhoB在肾透明细胞癌组织中的表达情况.选取60例肾透明细胞癌组织标本,详细收集病人临床病理资料,通过免疫组化方法检测RhoB在肾透明细胞癌组织中的蛋白表达水平,并分析RhoB的蛋白水平与临床病理资料的关系.结果:与对应瘤旁肾组织相比,RhoB的mRNA及蛋白表达水平在肾透明细胞癌组织标本中明显降低;RhoB的蛋白表达水平在不同年龄、性别组间差异无统计学意义(P>0.05),而在肿瘤直径大小、T分期、临床分期和组织分级间差异有显著统计学差异(P<0.05).结论:RhoB的表达降低可能在肾透明细胞癌的肿瘤发生中发挥作用,且其表达下降可能与肾透明细胞癌的恶性进展有关.     

A novel inflammation-associated prognostic signature for clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) are typically situated in a complex inflammatory and immune microenvironment, which has been reported to contribute to the unfavorable prognosis of patients with ccRCC. There would be beneficial clinical implications for elucidating the roles of its molecular characteristics in the inflammatory microenvironment. This is because it would facilitate the development of reliable biomarkers for pre-stratification prior to the designation of individualized treatment strategies. In the present study, RNA-sequencing data from 607 patients were retrospectively analyzed to elucidate the profile of inflammatory molecules. Based on this, an inflammatory prognostic signature (IPS) was developed and further validated using clinical ccRCC samples. Subsequently, the associated mechanisms in terms of the immune microenvironment and molecular pathways were then investigated. This proposed IPS was found to exhibit superior accuracy compared with the criterion of a good prognostic model for the prediction of patient prognosis from ccRCC [area under the receiver operating characteristic curve (AUC)=0.811] in addition to being an independent factor for prognostic risk stratification [hazard ratio: 11.73 (95% CI, 26.98-5.10); log-rank test, P<0.001]. Pathologically, ccRCC cells identified as high-risk according to their IPS presented with a more malignant tumor structure, including voluminous eosinophilic cytoplasm, acinar/lamellar/tubular growth patterns and atypic nuclei. High-risk ccRCC also exhibited higher infiltration levels by four types of immune cells, including T regulatory cells, but lower infiltration levels by mast cells. Pathways associated with immune-inflammation interaction, including the IL-17 pathway, were found to be upregulated in IPS-identified high-risk ccRCC. Furthermore, by combining the IPS with clinical factors, an integrated prognostic index was developed and validated for increasing the accuracy of patient risk-stratification for ccRCC (AUC=0.911). In conclusion, the complex regulatory mechanisms and molecular characteristics involved in ccRCC-inflammation interaction, coupled with their prognostic potential, were systematically elucidated in the present study. This may have important implications in furthering the understanding into the molecular mechanisms underlying this ccRCC-inflammation interaction, which can in turn be exploited for identifying high-risk patients with ccRCC prior to designing their clinical treatment strategy.     

Hepcidin as a prognostic biomarker in clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is a common malignancy of urologic neoplasms. Hepcidin is a pivotal modulator of iron metabolism involved in human cancers; however, the biological significance of hepcidin in ccRCC remains to be fully understood. Therefore, in this study, we evaluated the expression profiles of hepcidin in ccRCC from several public databases and found that hepcidin expression was upregulated in ccRCC, which was further validated in ccRCC cell lines, clinical samples, and tissue microarray (TMA) quantitative real-time PCR and immunohistochemistry. In addition, we found that the expression level of hepcidin was correlated with the age, T stage and pathologic stage of patients. Furthermore, hepcidin promoter methylation was significantly associated with the worse poor clinical parameters of ccRCC patients, and hepcidin was an independent prognostic factor. Mechanistically, enrichment analysis revealed that hepcidin participated in the immune-related and metabolism-related pathways. Hepcidin was positively correlated with not only immune infiltration and immune checkpoints but also tumor mutation burden and cytotoxic T lymphocyte. Finally, we validated the positive correlation of hepcidin with the marker of macrophage (CD68) in the TMA. Our findings provide insights into understanding the function and its underlying mechanism of hepcidin in ccRCC and suggest that hepcidin might serve as a potential predictive biomarker of response to immunotherapy and the prognosis of patients with ccRCC.     

肾透明细胞癌联合免疫治疗新策略——有氧糖酵解的研究进展及展望

肾恶性肿瘤的发病率逐年上升,其中肾透明细胞癌约占所有肾恶性肿瘤的80%,肾透明细胞癌独特的遗传背景和突变特征往往涉及以乏氧信号、糖酵解代谢、氨基酸代谢、线粒体氧化磷酸化等通路为代表的肿瘤微环境（tumor microenvironment,TME）内稳态失调。免疫检查点抑制剂（immune checkpoint inhibitor,ICI）联合酪氨酸激酶抑制剂（tyrosine kinase inhibitor,TKI）已经成为晚期肾透明细胞癌患者的一线治疗方案,但是,联合治疗方案的疗效仍有待提高,且缺乏明确诊断、指导用药、评估预后的生物标志物。近年来,多组学研究从不同层次探索肾透明细胞癌分子通路的异常改变。肾透明细胞癌发生代谢重编程,在氧气充足的情况下也以低效能的糖酵解为能量供应来源,促进自身无限生长,并且有氧糖酵解通路展现的显著异常与不良预后相关。肾透明细胞癌异常的糖酵解信号能促进肿瘤生长,并与TME中的免疫细胞相互作用,使促肿瘤免疫和抗肿瘤免疫平衡失调,造成抑制性免疫微环境,介导肿瘤免疫逃逸,从而对免疫治疗产生不利影响。因此,通过阻断异常糖代谢来抑制肿瘤生长,以有氧糖酵解通路和免疫微环境为切入点,可为肾透明细胞癌以及泛肿瘤治疗提供新的研究方向。然而,如何在复杂的肿瘤免疫微环境中最大程度地将肿瘤细胞代谢重编程转化为用药靶点并运用于临床实践仍待探讨。在肾透明细胞癌中,糖酵解抑制剂联合ICI或TKI作为新方案或能协同发挥抗肿瘤效应,逆转治疗抵抗。本文通过对糖酵解代谢途径中的关键限速酶、转运体及其抑制剂与肿瘤免疫微环境之间的关系进行综述,探讨糖酵解抑制剂在肾透明细胞癌中的作用机制和肿瘤免疫微环境的变化,及其与靶向治疗或免疫治疗联合应用的巨大临床转化价值,未来将为肾透明细胞癌的临床诊疗提供新思路,为患者带来临床获益。     

Clear cell renal cell carcinoma bone metastasis: What should be considered in prognostic evaluation

IntroductionKnowledge of clear cell renal cell carcinoma bone metastasis (ccRCC-BM) remains scarce. This study depicts clinical, pathological and outcome features of the disease and provides suggestions to establish prognosis prediction system more appropriate for ccRCC-BM.Materials and methodsPatients with ccRCC-BM had clinical, pathological data collected. Kaplan-Meier survival analysis was used for outcome profiles. Prognostic risks were evaluated using MSKCC/Motzer score. Univariate and multivariate logistic regression were performed to investigate association between clinical, pathological features and prognosis.ResultsIn the series containing 106 ccRCC-BM patients with 4:1 male predominance, 44.3% of them had synchronous bone metastasis and 28.3% had multi-organ metastasis. Axial bone was prone to bone metastasis and the incidence of severe skeletal-related events was 54.7%. Curative bone lesion resection was performed in 70.7% patients. The median overall survival (mOS) time was 45 months for all and 32 months for those in unfavorable risk stratification. Shorter time to bone metastasis (TTBM) [OR 1.019, 95% CI (1.007, 1.031)], elderly age [OR 1.040, 95% CI (1.001, 1.080)], concomitant multi-organ metastasis [OR 3.883, 95% CI (1.375, 10.967)] and carbonic anhydrase (CA)-IX expression loss [OR 58.824, 95% CI (2.653, 1000)] were associated with poor prognosis.ConclusionThe outcome of ccRCC-BM remained poor in unfavorable risk stratification. Bone lesion resection accompanied by systematic therapy for selected patient could improve prognosis. Shorter TTBM, elderly age, concomitant multi-organ metastasis and the expression loss of CA-IX along with gender-bias, feasibility for surgical treatment are suggested to be incorporated in modified ccRCC-BM-specific prognosis prediction system.     

透明细胞乳头状肾细胞癌7例报告并文献复习

背景与目的：透明细胞乳头状肾细胞癌是最近发现的一种少见的肾脏上皮源性恶性肿瘤,其生物学行为惰性,预后较好,该研究旨在分析透明细胞乳头状肾细胞癌的临床及病理学特点,与其他亚型鉴别,以免过度治疗。方法：收集7例透明细胞乳头状肾细胞癌病例,应用组织病理学、免疫组织化学法并结合相关文献分析其镜下及临床特点。结果：7例患者肿块均位于肾内,切面灰红或灰黄色,实性或囊实性,镜下肿瘤组织呈腺管状、微囊状或乳头状结构,肿瘤以一种结构为主或多种结构混合存在。细胞质透明,细胞核圆形或卵圆形,世界卫生组织（World Health Organization,WHO）/国际泌尿病理协会（International Society of Urological Pathology,ISUP）细胞核分级为1或2级。免疫表型：7例均表达CK7、CK8、vimentin、PAX-8、CA9和CK34βE12,Ki-67增殖指数为5%~10%,7例均不表达CD117、TFE3和CD10。对本组7例患者随访2个月至4年,均无复发及转移。结论：透明细胞乳头状肾细胞癌是一种少见的低度恶性肿瘤,形态学上与多种具有透明细胞和（或）乳头状细胞的肾癌有重叠,需要借助免疫组织化学进行鉴别。     

MSCT多期扫描在肾脏乏脂肪血管平滑肌脂肪瘤与透明细胞癌鉴别诊断中的应用价值

目的 探讨肾脏乏脂肪血管平滑肌脂肪瘤(fpAML)和透明细胞癌(ccRCC)多排螺旋CT(MSCT)平扫及多期增强扫描表现,提高诊断准确率.方法 选取2015年1月至2019年12月江苏省中医院经手术病理确诊的fpAML和ccRCC患者各20例,测量肿瘤的短径和长径、CT平扫及多期增强的肿瘤CT值及邻近肾皮质的CT值,...     

细胞骨架相关蛋白4在肾透明细胞癌中的表达及其临床意义

目的 探讨细胞骨架相关蛋白4(cytoskeleton-associated protein 4,CKAP4)在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)中的表达及其临床意义。方法 收集237例ccRCC患者的癌组织和癌旁组织,采用免疫组化法检测CKAP4蛋白的表达水平,采用χ²检验比较不同CKAP4蛋白表达水平患者的临床病理特征,Kaplan-Meier法和Log-rank检验分析不同CKAP4表达水平患者总生存期(overall survival,OS)和无进展生存期(progression-free survival,PFS)的差异,Cox比例风险回归分析影响ccRCC患者术后OS和PFS的因素。结果 ccRCC组织中CKAP4蛋白的高表达率显著高于癌旁组织(54.0%vs 23.6%,χ²=46.050,P<0.001)。Fuhrman分级高、肿瘤浸润深、有淋巴结转移、远处转移以及TNM分期高的ccRCC患者癌组织中CKAP4蛋白高表达(均P<0.05)。CKAP4蛋白高表达...     

环氧合酶-2、血管内皮生长因子在透明细胞性肾细胞癌中的表达及其与肿瘤发生、发展的关系

目的探讨环氧合酶-2（COX-2）、血管内皮生长因子（VEGF）表达与透明细胞性肾细胞癌（CCRCC）患者预后的关系。方法应用免疫组织化学EnVision法检测80例CCRCC患者肿瘤组织和20例输尿管癌或肾盂癌患者正常肾组织中COX一2、VEGF的表达情况,结合临床病理参数分析二者与CCRCC患者预后的关系。结果COX-2在CCRCC组织中的阳性表达率为65．00％（52／80）,显著高于正常肾组织中的10．00％（2／20）（x^2=7．760,P=0．021）;VEGF在CCRCC中的阳性表达率为61．25％（49／80）,高于正常肾组织中的20.00％（4／20）（X^2=8．870,P=0．012）。COX-2的表达和CCRCCTNM分期（X^2=8．200,P=0．005）、组织学分级（X^2=13．860,P=0．000）以及淋巴结转移（X^2=6．050,P=0．001）有相关性,而与年龄（X^2=0．560,P=0．663）和肿瘤大小（x^2=0．700,P=0．528）无相关性;CCRCC组织中COX-2和VEGF的表达呈正相关（r=0．485,P〈0．01）。COX-2、VEGF的表达和CCRCC患者预后相关（分别为X^2=18．280,P=0．038;X^2=6．420,P=0．042）。结论COX-2和VEGF在CCRCC组织中的表达与肿瘤的发生、发展及预后密切相关,可以作为CCRCC患者的预后指标。     

肾透明细胞癌组织Fibulin-3表达及其临床意义

目的 探讨Fibulin-3在肾透明细胞癌(clear cell renal cell carcinoma,CCRCC)中的表达及其临床意义.方法 收集中山大学附属第一医院2000-04-01-2007-04-01手术切除并经病理确诊的157例CCRCC患者的石蜡标本及相关临床病理资料.其中男10例,女56例,平均年龄51.7岁.制作组织芯片,应用免疫组织化学方法检测Fibulin-3蛋白的表达情况,根据评分结果将RCC患者分为Fibulin-3高表达和低表达两组,并分析其与患者多种临床参数的相关性.结果 Fibulin-3在157例CCRCC组织中的阳性表达率为100.0％,高表达58例,低表达99例.Fibulin-3蛋白表达与CCRCC患者的性别、年龄、肿瘤大小、Fuhrman分级和组织学类型等临床病理参数无相关性,P＞0.05.Kaplan-Meier生存分析显示,Fibulin-3高表达与肾癌患者的预后相关,P=0.003;多因素Cox回归分析表明,Fibulin-3是CCRCC患者生存时间的独立影响因素,HR=2.30,95％CI为1.278～4.140,P=0.005.结论 Fibulin-3高表达与CCRCC患者的不良预后相关,提示Fibulin-3是预测CCRCC患者预后的潜在分子标志.     

CIAPIN1 inhibits the growth and proliferation of clear cell renal cell carcinoma

Our previous studies indicated a direct correlation with loss of CIAPIN1 and carcinogenesis of tumor in human gastric cancer. Here we presented that the expression of CIAPIN1 was absent or significantly decreased in 102 cases of clear cell renal cell carcinoma (CCRCC) tissues (P < 0.05). Up-regulating CIAPIN1 by adenoviral vectors exhibited significant inhibition of CCRCC-derived cell growth in vitro and in vivo with G1 cell cycle arrest. Simultaneously, CIAPIN1-induced growth suppression was found partially to regulate various proteins, including inhibition of cyclinD1, cyclinE, cdk2, cdk4, p-Rb and VEGF, but up-regulation of p27^Kip1 and Rb.