Expression and clinical significance of PTPN12 in clear cell renal cell carcinoma

BackgroundClear cell renal cell carcinoma (ccRCC) is a common urological disease. Expression of the protein tyrosine phosphatase 12 gene (PTPN12) is decreased in many cancers; however, the relationship between PTPN12 gene function and renal cancer remains unclear.MethodsWe detected PTPN12 protein expression in ccRCC and corresponding normal tissues from 64 patients with ccRCC by immunohistochemistry, and relative PTPN12 mRNA levels by real-time quantitative polymerase chain reaction. The relationships between the relative expression levels of PTPN12 mRNA and the patients’ clinical data were analyzed.ResultsPTPN12 protein and mRNA expression levels were significantly lower in ccRCC compared with the corresponding normal tissues. The mRNA expression levels in the ccRCC and corresponding normal tissues from the 64 patients with ccRCC were 0.459±0.445 and 1.001±0.128, respectively, compared with the control (glyceraldehyde 3-phosphate dehydrogenase). There was a significant correlation between relative expression of PTPN12 mRNA in ccRCC tissues and tumor diameter and clinical stage.ConclusionThe expression levels of PTPN12 protein and mRNA were significantly lower in ccRCC tissues compared with normal tissues. The role of PTPN12 may provide new insights and evidence to aid the diagnosis and targeted therapy of ccRCC.     

Misbehaviour of XIST RNA in Breast Cancer Cells

A role of X chromosome inactivation process in the development of breast cancer have been suggested. In particular, the relationship between the breast cancer predisposing gene BRCA1 and XIST, the main mediator of X chromosome inactivation, has been intensely investigated, but still remains controversial. We investigated this topic by assessing XIST behaviour in different groups of breast carcinomas and in a panel of breast cancer cell lines both BRCA1 mutant and wild type. In addition, we evaluated the occurrence of broader defects of heterochromatin in relation to BRCA1 status in breast cancer cells. We provide evidence that in breast cancer cells BRCA1 is involved in XIST regulation on the active X chromosome, but not in its localization as previously suggested, and that XIST can be unusually expressed by an active X and can decorate it. This indicates that the detection of XIST cloud in cancer cell is not synonymous of the presence of an inactive X chromosome. Moreover, we show that global heterochromatin defects observed in breast tumor cells are independent of BRCA1 status. Our observations sheds light on a possible previously uncharacterized mechanism of breast carcinogenesis mediated by XIST misbehaviour, particularly in BRCA1-related cancers. Moreover, the significant higher levels of XIST-RNA detected in BRCA1-associated respect to sporadic basal-like cancers, opens the possibility to use XIST expression as a marker to discriminate between the two groups of tumors.     

E2F⁃3、E2F⁃4蛋白在透明细胞性肾细胞癌中的表达及临床意义

目的通过检测E2F3、E2F4蛋白在透明细胞性肾细胞癌(ccRCC)及相应癌旁组织中的表达情况,探讨E2F3、E2F4蛋白与ccRCC发生发展的关系。方法采用免疫组织化学SP法检测147例ccRCC及相应癌旁组织中E2F3、E2F4蛋白的表达情况,进一步分析与ccRCC患者临床病理特征的关系。结果E2F3蛋白在ccRCC组织中的阳性率(76.19%)明显高于癌旁组织(18.37%),差异具有显著统计学意义(P<0.001)。E2F4在ccRCC组织中的阳性率(68.71%)明显高于相应的癌旁组织阳性率(28.57%),差异具有显著统计学意义(P<0.001)。E2F3的表达强度与WHO/ISUP分级、临床分期、肿瘤大小、肿瘤性坏死、肾静脉内癌栓相关(P<0.05);而与年龄、性别、有无淋巴结转移、有无远处转移无关(P>0.05)。E2F4的表达强度与WHO/ISUP分级、肿瘤大小、肾静脉内癌栓相关(P<0.05);而与年龄、性别、临床分期、有无淋巴结转移、远处转移、肿瘤性坏死无关(P>0.05)。E2F3、E2F4蛋白表达的相关性检验显示,二者表达无明显相关性(rs=0.105,P=0.206)。结论E2F3、E2F4可能参与了ccRCC的发生及进展,二者表达无明显协同作用。     

精囊蛋白SG1在肾透明细胞癌中的表达和预后意义

目的探讨精囊蛋白SG1在肾透明细胞癌患者癌旁组织和癌组织中的表达差异,结合临床指标,研究其临床意义。方法以qRT-PCR检测24例明确诊断为肾透明细胞癌患者的癌组织和癌旁组织中的SG1表达水平,以免疫组化研究53例肾透明细胞癌标本中的SG1蛋白表达水平,并进一步研究其临床意义。结果 qRT-PCR显示癌组织中的SG1水平明显低于癌旁组织的(P<0.01)。53例癌组织中,抗SG1免疫组化染色30例阳性(57%),明显低于对应癌旁组织的全部阳性(100%)。SG1表达量和肾透明细胞癌临床分期呈负相关(pT1² vs.pT32 vs.pT3⁴,P<0.000 1),且SG1表达阴性的患者复发概率升高(P<0.01)。结论 SG1在肾透明细胞癌中为低表达,SG1可预测肾透明细胞癌的进展和预后。     

Fanconi anemia signaling network regulates the spindle assembly checkpoint

Fanconi anemia (FA) is a heterogenous genetic disease with a high risk of cancer. The FA proteins are essential for interphase DNA damage repair; however, it is incompletely understood why FA-deficient cells also develop gross aneuploidy, leading to cancer. Here, we systematically evaluated the role of the FA proteins in chromosome segregation through functional RNAi screens and analysis of primary cells from patients with FA. We found that FA signaling is essential for the spindle assembly checkpoint and is therefore required for high-fidelity chromosome segregation and prevention of aneuploidy. Furthermore, we discovered that FA proteins differentially localize to key structures of the mitotic apparatus in a cell cycle–dependent manner. The essential role of the FA pathway in mitosis offers a mechanistic explanation for the aneuploidy and malignant transformation known to occur after disruption of FA signaling. Collectively, our findings provide insight into the genetically unstable cancers resulting from inactivation of the FA/BRCA pathway.     

Non‐invasive urine markers for the differentiation between RCCs and oncocytoma

BackgroundRecently, our group showed that Vim3 is overexpressed in tissue samples of renal oncocytomas and Mxi‐2 in clear cell renal carcinoma (ccRCC). The mechanism leading to the truncation of both proteins is known and involves with two miRs, both detectable in urine. Since the analysis of miRs is time‐consuming, our aim was to identify the truncated proteins in urine instead. Furthermore, urine samples from small renal masses (SRMs) (n = 45, <4 cm) were analyzed to get a pre‐surgical differentiation of the cancer subtypes.MethodsUrines were accessed from the urological biobank (n = 350). Proteins were isolated from urine samples, and Western blots were performed. Each sample was analyzed with ELISA for the expression of Vim3 and Mxi‐2. A lateral flow assay was established. For the detection of SRMs, the miRs were isolated and qRT‐PCR was performed.ResultsA significant increase of Vim3 in urines from patients with oncocytoma (n = 20) was detectable with ELISA compared to all other subtypes of RCCs (chromophobe (n = 50), papillary (n = 40), ccRCC (n = 200), and controls (n = 40) (***p < 0.0001)). Mxi‐2 was predominantly overexpressed in ccRCCs (***p < 0.0001). Lateral flow assay of Vim3 and Mxi‐2 shows two bands in the case of oncocytoma and ccRCC indicating the specificity of this test.For SRMs, an overexpression of miR‐15a/Mxi2 was detectable in urine samples from ccRCC and chromoRCC patients. In contrast to that, miR‐498/Vim3 were predominantly overexpressed in oncocytoma patients.ConclusionBoth proteins (Vim3 and Mxi‐2) were detectable in patients’ urines and can be used for the non‐invasive differentiation of kidney cancers.     

HSP60和NRF2在透明细胞性肾细胞癌中的表达及临床价值

目的 研究透明细胞性肾细胞癌(ccRCC)中热休克蛋白60(HSP60)及核因子E2相关因子2(NRF2)的表达及临床意义.方法 选取2017年3月至2019年3月在安康市中心医院诊治的89例ccRCC患者为研究对象,应用荧光定量聚合酶链反应(qPCR)及免疫组织化学检测癌组织及癌旁组织中HSP60、NRF2的表达.统...     

TSP-1在肾透明细胞癌与外周血中的表达及临床意义

目的 检测并分析凝血酶敏感蛋白-1(thrombospondin-1,TSP-1)在肾透明细胞癌患者肾癌、癌旁组织、正常肾组织及外周血中的表达及临床意义.方法 收集47例肾透明细胞癌患者的标本包括:47例肾癌组织、41例癌旁组织、38例正常肾组织、35例患者外周血标本,制备组织抽提液及外周血清.运用ELISA法检测三种组织及血清中TSP-1的表达,并结合临床资料分析其临床意义.结果 TSP-1在肾癌组织、癌旁组织、正常肾组织三者间的表达水平分别为36.44±4.80ng/mL、42.55±4.12ng/mL、46.55±1.55ng/mL,依次递增,三种组织间的表达水平,两两比较均有统计学差异(P均＜0.01).TSP-1在肾癌组织中的表达,随着肾癌TNM分期与Fuhrman分级的提高而均降低,且都呈负相关(r=-0.84,P＜0.01;r=-0.70,P＜0.01).结论 TSP-1在肾透明细胞癌组织中低表达,可能在抑制肿瘤血管形成过程中起重要作用;TSP-1在肾透明细胞癌组织中的表达与肾癌的TNM分期和Fuhrman分级关系密切,可作为评判肾透明细胞癌发展及恶性程度的指标.     

Visfatin and global histone H3K9me levels in colon cancer

BackgroundVisfatin is considered to be a biomarker in various types of cancers, including colon cancer. Moreover, evidence for epigenetic mechanism must be reported for an association between visfatin level and colon cancer. Therefore, this study was designed to investigate the status of visfatin expression and the global histone three modifications in colon cancerous tissue.MethodsColon cancerous tissue and paired adjacent non-cancerous tissue from 30 patients were used to determine the global histone three modifications using Western blot. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to assess visfatin''s expression level in tissues.ResultsThe visfatin and the global H3K9me expression levels were significantly higher in colon cancerous tissue than in the paired adjacent non-cancerous tissue.ConclusionThe present study makes a crucial noteworthy contribution to visfatin effect on colon cancer development via H3K9me.     

LINC01232 serves as a novel biomarker and promotes tumour progression by sponging miR-204-5p and upregulating RAB22A in clear cell renal cell carcinoma

BackgroundLong non-coding RNAs (lncRNAs) are involved in the progression of various cancers, including clear cell renal cell carcinoma (ccRCC). This study aimed to investigate the expression and prognostic value of long intergenic non-protein coding RNA (LINC) 01232 in ccRCC and preliminary explore the molecular mechanism underlying the role of LINC01232 in ccRCC progression.MethodsTumour tissues and adjacent normal tissues of 122 patients with ccRCC were collected in this study. The levels of LINC01232, microRNA (miR)-204-5p and RAB22A were measured by quantitative real-time PCR. The proliferation, migration and invasion of ccRCC cells were detected by cell counting kit-8 assay and Transwell assay, respectively. The interaction among LINC01232, miR-204-5p and RAB22A was confirmed by bioinformatics analysis, dual-luciferase reporter assay and Pearson correlation analysis. The association of LINC01232 and miR-204-5p with ccRCC patient survival was verified by the Kaplan–Meier method and log-rank test. The prognostic value of LINC01232 in ccRCC was confirmed by Cox regression analysis.ResultsLINC01232 expression was increased in ccRCC tumour tissues and ccRCC cells and independently predicted the prognosis of ccRCC patients. In addition, LINC01232 silencing inhibited ccRCC cell proliferation, migration and invasion. Moreover, LINC01232 served as a sponge for miR-204-5p, and miR-204-5p reduction reversed the inhibitory effect of LINC01232 silencing on ccRCC cell function. Furthermore, LINC01232 could sponge miR-204-5p, causing the elevation of RAB22A in ccRCC, thereby promoting ccRCC cell function.ConclusionLINC01232 may be an independent prognostic biomarker in ccRCC and plays an oncogenic role in ccRCC progression by sponging miR-204-5p and upregulating RAB22A.     

Genetic and epigenetic markers to identify high risk patients for multiple early gastric cancers after treatment with endoscopic mucosal resection

The development of multiple gastric cancer is a major problem after the endoscopic resection of the first early gastric cancer. To find out markers to identify high risk patients, we analyzed the microsatellite instability (MSI) status and hypermethylation of tumor-related genes in multiple gastric cancers. Sixty-four adenocarcinomas resected by endoscopy, including 32 early solitary gastric cancers (SGCs) from 32 patients and 32 multiple gastric cancers (MGCs) from 14 patients, were employed. We analyzed MSI and the methylation status of promoter regions of the hMLH1, MGMT, p16 and E-cadherin using methylation-specific Polymerase Chain Reaction. Expression levels of hMLH1 were examined by immunohistochemistry. MSI (+) was detected in 5 of the 14 (35.7%) patients with MGCs, and in only 3 of the 32 patients (9.3%) with SGCs. Significant differences were observed between the 2 groups (p<0.001). Hypermethylation of hMLH1 was more frequently detected in MGCs than in SGCs (p<0.01), whereas significant difference was not observed in the frequency of MGMT, p16 or E-cadherin promoter methylation between the 2 groups. In conclusion, our results indicate that inactivation of hMLH1 through promoter hypermethylation may be involved in the development of multiple gastric cancers following the MSI pathway.     

Amphiregulin Promotes Resistance to Gefitinib in NonSmall Cell Lung Cancer Cells by Regulating Ku70 Acetylation

Multiple molecular resistance mechanisms reduce the efficiency of receptor tyrosine kinase inhibitors such as gefitinib in non-small cell lung cancer (NSCLC). We previously demonstrated that amphiregulin (Areg) inhibits gefitinib-induced apoptosis in NSCLC cells by inactivating the proapoptotic protein BAX. In this part of the investigation, we studied the molecular mechanisms leading to BAX inactivation. We show that Areg prevents gefitinib-mediated acetylation of Ku70. This augments the BAX-Ku70 interaction and therefore prevents BAX-mediated apoptosis. Accordingly, Areg or Ku70 knock down restore BAX activation and apoptosis in gefitinib-treated H358 cells in vitro. In addition, overexpression of the histone acetyltransferase (HAT) CREB-binding protein (CBP) or treatments with histone deacetylase (HDAC) inhibitors sensitize H358 cells to gefitinib. Moreover, a treatment with vorinostat, a HDAC inhibitor strongly sensitized tumors to gefitinib in vivo. These findings suggest new prospects in combining both HDAC and epidermal growth factor receptor inhibitors for the treatment of NSCLC.     

探究PODXL在肾透明细胞癌中的表达及相关意义

目的：研究PODXL在透明细胞肾细胞癌(ccRCC)组织与正常组织中的表达差异,并初步分析PODXL对肾透明细胞癌的临床诊断和患者预后的关系。实验方法：运用癌症基因组图谱(TCGA)数据库收集RNA-Seq数据和临床数据,观察ccRCC患者组织与正常组织中PODXL基因表达量的差异,同时分析PODXL与临床病理学之间的关系及与预后之间的关系。结果：在TCGA数据库中总体526例ccRCC样本的癌和癌旁组织中的PODXL的表达水平分别为6.42±1.42和3.61±1.31,差异有统计学意义(P<0.001)。72例配对的ccRCC和癌旁组织中的PODXL表达水平分别为6.55±1.23和3.53±1.35,有统计学意义(P<0.001)。PODXL的表达在患者性别、肾肿瘤分级、T分期、M分期、临床分期等均有不同差异(P<0.001)。生存分析发现PODXL低表达量组和高表达量组的生存时间分别为(1446.75±956.32)d和(1253.81±939.57)d,差异有统计学意义(Log-rank=14.38,P<0.001)。ROC曲线结果显示,PODXL基因可作为一个灵敏度和特异度较高的ccRCC诊断指标：其中AUC为0.805,敏感性为67.26%,特异性为78.17%。结论：PODXL在ccRCC中高表达与临床病理特征呈正相关,是预后不良因素,有望作为肾透明细胞癌患者的生存预后及诊断的潜在生物学标志。     

The telomerase inhibitor PinX1 is a major haploinsufficient tumor suppressor essential for chromosome stability in mice

Telomerase is activated in most human cancers and is critical for cancer cell growth. However, little is known about the significance of telomerase activation in chromosome instability and cancer initiation. The gene encoding the potent endogenous telomerase inhibitor PinX1 (PIN2/TRF1-interacting, telomerase inhibitor 1) is located at human chromosome 8p23, a region frequently exhibiting heterozygosity in many common human cancers, but the function or functions of PinX1 in development and tumorigenesis are unknown. Here we have shown that PinX1 is a haploinsufficient tumor suppressor essential for chromosome stability in mice. We found that PinX1 expression was reduced in most human breast cancer tissues and cell lines. Furthermore, PinX1 heterozygosity and PinX1 knockdown in mouse embryonic fibroblasts activated telomerase and led to concomitant telomerase-dependent chromosomal instability. Moreover, while PinX1-null mice were embryonic lethal, most PinX1+/- mice spontaneously developed malignant tumors with evidence of chromosome instability. Notably, most PinX1 mutant tumors were carcinomas and shared tissues of origin with human cancer types linked to 8p23. PinX1 knockout also shifted the tumor spectrum of p53 mutant mice from lymphoma toward epithelial carcinomas. Thus, PinX1 is a major haploinsufficient tumor suppressor essential for maintaining telomerase activity and chromosome stability. These findings uncover what we believe to be a novel role for PinX1 and telomerase in chromosome instability and cancer initiation and suggest that telomerase inhibition may be potentially used to treat cancers that overexpress telomerase.     

Heterochromatin Protein 1 (HP1) Proteins Do Not Drive Pericentromeric Cohesin Enrichment in Human Cells

Sister chromatid cohesion mediated by cohesin is essential for accurate chromosome segregation. Classical studies suggest that heterochromatin promotes cohesion, but whether this happens through regulation of cohesin remains to be determined. Heterochromatin protein 1 (HP1) is a major component of heterochromatin. In fission yeast, the HP1 homologue Swi6 interacts with cohesin and is required for proper targeting and/or stabilization of cohesin at the centromeric region. To test whether this pathway is conserved in human cells, we have examined the behavior of cohesin in cells in which the levels of HP1 alpha, beta or gamma (the three HP1 proteins present in mammalian organisms) have been reduced by siRNA. We have also studied the consequences of treating human cells with drugs that change the histone modification profile of heterochromatin and thereby affect HP1 localization. Our results show no evidence for a requirement of HP1 proteins for either loading of bulk cohesin onto chromatin in interphase or retention of cohesin at pericentric heterochromatin in mitosis. However, depletion of HP1gamma leads to defects in mitotic progression.     

B/A Measurement of Clear Cell Renal Cell Carcinoma versus Healthy Kidney Tissue

The acoustic parameter of non-linearity B/A has been found capable of discriminating some types of pathological tissue from healthy tissue. The literature on the utility of B/A for cancer diagnostics is very limited, with measurements on the human breast and liver. This work expands the current research on cancer diagnostics by B/A assessment of eight slices of human clear cell renal cell carcinoma (ccRCC) from two patients and four slices of healthy kidney tissue from two healthy kidney samples. The Wilcoxon test identified the B/A distribution of malignant tissue as not significantly different from that of healthy tissue. An alternative way of defining outliers resulted in median B/A values of 8.1 for ccRCC and 6.8 for healthy tissue (p < 0.05). Acoustic attenuation at 2.1 MHz was significantly greater (p < 0.05) for ccRCC (1.7 dB/cm) than for healthy tissue (1.0 dB/cm). The observed differences in the measured values suggest that B/A and acoustic attenuation may represent potential diagnostic markers of ccRCC. More data and an improved experimental design are required to provide a definitive conclusion on the utility of B/A for cancer diagnostics.     

Epigenetic biomarkers in colorectal cancer diagnostics

Colorectal cancer (CRC) is a significant health burden worldwide. Despite advancements in treatment options, improvements in CRC patient survival have been limited owing to lack of early detection and limited capacity for optimal therapeutic decision-making. Biomarkers to improve CRC diagnosis, prognosis and prediction of treatment response therefore represent opportunities to improve patient outcome. In addition to genetic alterations and genomic instability, it is now clear that epigenetic alterations play dramatic roles in driving tumor onset and progression in CRC. A recent surge in investigation of epigenetic biomarkers including DNA methylation, miRNA expression and histone modifications has demonstrated that these alterations may be enticing translational biomarker candidates in CRC. In particular, methylation kits have already been incorporated into clinical practice for a handful of cancers, including CRC. This review will aim to summarize the established and emerging roles of epigenetic modifications in CRC detection, prognostication and prediction of treatment response.     

Chromosomal instability detected by fluorescence in situ hybridization in Japanese breast cancer patients.

The relationship between chromosomal instability (CIN) and prognostic factors was investigated in 31 breast cancers and 5 benign breast lesions (three fibroadenomas and two papillomas). Using fluorescence in situ hybridization (FISH) with chromosome-specific DNA probes of chromosomes 1, 2, 6, 7, 10, 11, 17 and 18, CIN for each case was determined. CIN varied from 8.1% to 59.3% among the breast cancer patients tested, and was significantly higher than that observed in the benign breast lesions (p<0.01). Moreover, CIN showed a significant correlation with lymph node metastases (p<0.05) and estrogen receptor negativity (p<0.01). These findings suggest that CIN might be useful in the prediction of the biological aggressiveness of breast cancers.