An Update on MDMX and Dual MDM2/X Inhibitors.

MicroRNA-520c (miR-520c) and microRNA-373 (miR-373) are originally characterized as both oncogenes and tumor suppressors in different types of human cancers. In this study, we found that translation of mRNA of MT1-MMP, an oncogene related to tumor metastasis, was well inhibited by miR-520c and miR-373 in several types of human cancer cells. Our experimental data demonstrated that these two microRNAs inhibited the translation of mRNA of MT1-MMP and down-regulated its proteolytic enzyme activities via targeting 3'UTR of mRNA of MT1-MMP, further decreased activating proMMP2 into active MMP2 in fibrosarcoma HT1080, benign prostatic hyperplasia epithelial cell BPH-1 and glioblastoma U87GM. More interestingly, from the effects of microRNAs on cell functions, we found that cell growth were all blocked on fibronectin and type IV collagen coated plates and also in three-dimension type I collagen lattice but enhanced only in HT1080 cells on type IV collagen coated plates and in three-dimension type I collagen lattice; cell migration results showed the same effect as that of cell growth. The difference was due to up-regulating the expression of MMP9 gene by miR-520c and miR-373 in HT1080 cells but not in BPH-1 and U87GM cells. Our findings suggest that miR-520c and miR-373, which have different roles in different type of cancer via regulating the translation of mRNA of MT1-MMP and the expression of MMP9 gene, might have an important clue on clinic when selecting the therapeutic regimen and finding new drugs for intervention in different kinds of cancer.

Simple SummaryDespite significant advances in treatment, breast cancer continues to be prevalent around the world. Metastasis associated in colon cancer 1 (MACC1) has been shown to be involved in the progression of more than 20 different types of cancer, including breast cancer, and performs specific functions in different cancers. MACC1’s biological role, molecular mechanism, and pathway are important for understanding cancer progression. MACC1 is highly expressed in breast cancer and has been linked to metastasis, staging, and a poor prognosis. This review article attempts to elucidate the prognostic value of MACC1 expression in breast cancer, its role in immune and anti-radiation therapy, and the network regulatory mechanisms involved, and discusses the application of MACC1 in the treatment and prevention of breast cancer. It is hoped that the use of MACC1 as a biomarker in breast cancer surveillance and clinical guidance can be promoted.Metastasis associated in colon cancer 1 (MACC1) is an oncogene first identified in colon cancer. MACC1 has been identified in more than 20 different types of solid cancers. It is a key prognostic biomarker in clinical practice and is involved in recurrence, metastasis, and survival in many types of human cancers. MACC1 is significantly associated with the primary tumor, lymph node metastasis, distant metastasis classification, and clinical staging in patients with breast cancer (BC), and MACC1 overexpression is associated with reduced recurrence-free survival (RFS) and worse overall survival (OS) in patients. In addition, MACC1 is involved in BC progression in multiple ways. MACC1 promotes the immune escape of BC cells by affecting the infiltration of immune cells in the tumor microenvironment. Since the FGD5AS1/miR-497/MACC1 axis inhibits the apoptotic pathway in radiation-resistant BC tissues and cell lines, the MACC1 gene may play an important role in BC resistance to radiation. Since MACC1 is involved in numerous biological processes inside and outside BC cells, it is a key player in the tumor microenvironment. Focusing on MACC1, this article briefly discusses its biological effects, emphasizes its molecular mechanisms and pathways of action, and describes its use in the treatment and prevention of breast cancer.

MicroRNAs (miRNAs) are a group of small non-coding RNAs that post-transcriptionally control expression of genes by targeting mRNAs. miRNA alterations partake in the establishment and progression of different types of human cancer. Consequently, expression profiling of miRNA in human cancers has correlations with cancer detection, staging, progression, and response to therapies. Particularly, amplification, deletion, abnormal pattern of epigenetic factors and the transcriptional factors that mediate regulation of primary miRNA frequently change the landscape of miRNA expression in cancer. Indeed, changes in the quantity and quality of miRNAs are associated with the initiation of cancer, its progression and metastasis. Additionally, miRNA profiling has been used to categorize genes that can affect oncogenic pathways in cancer. Here, we discuss several circulating miRNA signatures, their expression profiles in different types of cancer and their impacts on cellular processes.

Chromosome 13q22.1 has previously been identified to be a susceptibility locus for pancreatic cancer in Chinese and European ancestry populations. This pleiotropy study aimed to identify novel variants in this region associated with susceptibility to different types of human cancer. To fine-map the 13q22.1 region, imputation analyses were conducted on the basis of the GWAS data of 2,031 esophageal squamous cell cancer (ESCC) cases and 2,044 controls and 5,930 SNPs (625 directly genotyped and 5,305 well imputed). Promising associations were then examined in ESCC (4,146 cases and 4,135 controls), gastric cardia cancer (1,894 cases and 1,912 controls), noncardia gastric cancer (1,007 cases and 2,243 controls), and colorectal cancer (1,111 cases and 1,138 controls). Fine mapping and biochemical analyses were further performed to elucidate the potential function of novel variants. Two novel variants, rs1924966 and rs115797771, were associated with ESCC risk (P = 1.37 × 10(-10) and P = 2.32 × 10(-10), respectively) and were also associated with risk of gastric cardia cancer (P = 0.0003 and P = 0.0018, respectively) but not gastric cancer and colorectal cancer. Fine-mapping revealed another SNP, rs58090485, in strong linkage disequilibrium with rs115797771 (r(2) = 0.94). Functional analysis showed that this SNP disturbs a transcriptional repressor binding to the promoter region of KLF5, which might result in high constitutional expression of KLF5. These results demonstrate that variants mapped on 13q22.1 are associated with the risk of different types of cancer. 13q22.1 might serve as a biomarker for the identification of individuals at risk for ESCC and gastric cardia cancer.

Cystatin E/M (CST6), a representative cysteine protease inhibitor, plays both tumor-promoting and tumor-suppressing functions and is pursued as an epigenetically therapeutic target in special cancer types. However, a comprehensive and systematic analysis for CST6 in pan-cancer level is still lacking. In the present study, we explored the expression pattern of CST6 in multiple cancer types across ∼10,000 samples from TCGA (The Cancer Genome Atlas) and ∼8,000 samples from MMDs (Merged Microarray-acquired Datasets). We found that the dynamic expression alteration of CST6 was consistent with dual function in different types of cancer. In addition, we observed that the expression of CST6 was globally regulated by the DNA methylation in its promoter region. CST6 expression was positively correlated with the epithelial cell infiltration involved in epithelial-to-mesenchymal transition (EMT) and proliferation. The relationship between CST6 and tumor microenvironment was also explored. In particular, we found that CST6 serves a protective function in the process of melanoma metastasis. Finally, the clinical association analysis further revealed the dual function of CST6 in cancer, and a combination of the epithelial cell infiltration and CST6 expression could predict the prognosis for SKCM patients. In summary, this first CST6 pan-cancer study improves the understanding of the dual functional effects on CST6 in different types of human cancer.

Cancer is one of the larger families of diseases; it is a collection of 100s of diseases that involve abnormal cell to grow and spread to the other parts of the body which leads to worldwide death of the human being. There are different types of cancer, so for those we need to identify and classify all those different types. In the field of bioinformatics, the main important thing is cancer diagnosis, so we need to do it by selecting the subset of feature gene. In cancer diagnosis, the greatest significance is classifying the different types of tumors. By providing an accurate prediction for various types of tumors, we can provide a better treatment for cancer patients and also it reduces the toxicity on patients. The main important thing in this paper is to differentiate between cancer subtypes by creating different methodologies. This paper explains different methodologies, based on gene profiling for the classification and prediction for different types of human cancer. The proposed methodology in this paper is a combination of symmetrical uncertainty (SU) and genetic algorithm (GA).

Introduction Annexin A11 was previously identified as an autoantigen in 4.1–10.1% of patients with various systemic autoimmune diseases. In this study, an enzyme-linked immunosorbent assay (ELISA) was developed to investigate the occurrence and features of anti-annexin A11 autoantibodies in sera from patients with different types of cancer. Methods The recombinant protein of GST fused to the N-terminal domain (1–175 residues) of human annexin A11 was expressed and used as antigen in ELISA. A total of 246 serum specimens were analyzed, which includes sera from healthy women (77), patients with ovarian cancer (72), breast cancer (18), colon cancer (19), pancreatic cancer (20), prostate cancer (20), and diabetes (20). Results The overall titer of anti-annexin A11 autoantibodies in ovarian cancer patients (or primary tumors only) was found much higher than that in healthy controls (P < 0.05). At the cut-off value designating positive reaction, anti-annexin A11 autoantibodies were detected in 12.5% (5/40) of primary ovarian cancer patients with a significant difference from 2.6% (2/77) of the healthy controls (P < 0.05), but only in 6.25% (2/32) of recurrent tumors. ROC curve demonstrated the potential diagnostic value of anti-annexin A11 autoantibodies in primary ovarian cancer patients with an AUC of 0.62 (0.52–0.73). Anti-annexin A11 autoantibodies were also detected in 5.26% (1/19) of colon cancer and 10% (2/20) of diabetes patients but without significant difference from the healthy controls. Conclusion A convenient assay to detect anti-annexin A11 autoantibodies in patients was developed, and the experimental data are promising but need to be expanded to address their biological/clinical relevance.

Tumor-specific promoter hypermethylation of large tumor suppressor, homolog 2 (LATS2), a tumor suppressor gene, has been investigated using methylation-specific polymerase chain reaction (MSP) assays in different types of human cancer producing conflicting results. The aim of the present study was to evaluate the methylation status of the LATS2 promoter region using bisulfite sequencing with a next generation sequencer for breast cancer. In the 11 patients enrolled in the present study, the LATS2 promoter methylation index (MI) was uniformly high in tumor and normal tissues of the breast (median, 84.0 and 87.4%, respectively). The presence of LATS2 promoter hypermethylation was confirmed in isolated tumor cells and normal epithelial cells using the magnetic-activated cell sorting method. In situ hybridization for LATS2 messenger RNA (mRNA) revealed that the mRNA expression of LATS2 was higher in normal epithelial cells, compared with tumor cells, however, it was not significantly associated with LATS2 MI. In 12 breast cancer cell (BCC) lines and two normal breast cell lines, the LATS2 promoter was uniformly hypermethylated with no correlation between the mRNA expression of LATS2 and the LATS2 MI. In addition, treatment of the BCC lines with a demethylating reagent had minimal effect on the mRNA expression of LATS2 in any of these cell lines. These results demonstrated that LATS2 hypermethylation was not involved in silencing the mRNA expression of LATS2 mRNA. The lower mRNA expression level of LATS2 in tumor cells, compared with normal epithelial cells, suggested the possible involvement of downregulation in the mRNA expression of LATS2 in the pathogenesis of breast cancer. Therefore, the conflicting results previously reported for LATS2 promoter methylation in different types of cancer, detected using MSP assays may be attributable to the low fidelity of the MSP assay.

Focus on kidney cancer

背景与目的肺癌是目前世界上发病率和死亡率较高的恶性肿瘤之一。然而，肺癌进展的确切机制仍不清楚。肿瘤坏死因子受体相关因子（tumor necrosis factor receptor associated factor, TRAF）家族成员是胞质接头蛋白，既行使接头蛋白的功能又具有泛素连接酶活性，可调控多种受体信号途径，进而激活核因子κB（nuclear factor kappa-B, NF-κB）、有丝分裂原活化蛋白激酶（mitogen-activated protein kinase, MAPK）和干扰素调节因子（interferon regulatory factor, IRF）信号。本研究旨在探讨TRAFs在不同组织和癌症类型中的表达以及在非小细胞肺癌（non-small cell lung cancer, NSCLC）中的mRNA表达、蛋白表达、预后意义及功能富集分析，为NSCLC的诊断和治疗提供新策略。方法利用基因型组织表达数据库的RNA测序数据集分析TRAF家族成员在不同人体组织中的表达水平；利用癌细胞百科全书数据库中的RNA测序数据集分析TRAF家族成员在不同类型癌症细胞系中的表达水平；利用癌症基因组图谱（The Cancer Genome Atlas, TCGA）数据库的RNA测序数据集分析人类不同类型癌症中TRAF家族成员的mRNA表达水平；利用人类蛋白质图谱数据库中的免疫组化数据分析NSCLC包括肺腺癌和肺鳞癌中的TRAF家族成员的蛋白表达水平；利用Kaplan-Meier Plotter中的NSCLC患者的生存数据集，通过对数秩检验评估TRAF家族成员的表达与预后生存的相关性；利用TCGA数据库中NSCLC患者的RNA测序数据集对TRAF家族成员相关基因进行基因本体论功能注释分析和京都基因与基因组百科全书通路富集分析；利用基于TCGA数据库的RNA测序数据集，通过ESTIMATE对TRAF家族成员的表达水平与肿瘤免疫微环境进行相关性分析。结果TRAF家族成员呈现显著的组织表达特异性，TRAF2、TRAF3、TRAF6和TRAF7广泛表达于多种组织，而TRAF1、TRAF4和TRAF5则在个别组织中表达受限；TRAF家族成员的表达在不同类型癌症细胞系中具有高度特异性；在肺腺癌和肺鳞癌组织的mRNA数据中，TRAF2、TRAF4、TRAF5和TRAF7的表达均上调，TRAF6的表达均下调，TRAF1仅在肺腺癌中上调，而TRAF3仅在肺鳞癌中上调；除TRAF3、TRAF4和TRAF7外，其他TRAF蛋白在肺腺癌和肺鳞癌组织中的免疫组化染色显著加深；TRAF2、TRAF4和TRAF7的表达与肺癌患者的总生存期呈负相关，TRAF3、TRAF5和TRAF6的表达与肺癌患者的总生存期呈正相关，而TRAF1的表达与肺癌患者的总生存期无明显相关性；TRAF家族成员差异性调控NF-κB、免疫应答、细胞黏附及RNA剪接相关的等多条信号通路；TRAF家族成员的表达水平与肿瘤免疫微环境中的免疫细胞浸润和基质细胞含量密切相关，且不同成员之间的相关性表现出正负差异。结论TRAF家族成员在不同组织和癌症类型中表现出特异性差异表达，NSCLC中多数TRAF家族成员在mRNA和蛋白水平表达上调，其中，高表达的TRAF2、TRAF4和TRAF7提示预后不良，TRAF家族成员参与炎症、免疫、黏附及代谢等多条通路，并影响肿瘤免疫微环境。

Chapter 232 - Ras and Cancer

The miR-200 Family: Central Player for Gain and Loss of the Epithelial Phenotype

Hsp90 is a ubiquitous, ATP-dependent chaperone, essential for eukaryotes. It possesses a broad spectrum of substrates, among which is the p53 transcription factor, encoded by a tumor-suppressor gene. Here, we elucidate the role of the adenine nucleotide in the Hsp90 chaperone cycle, by taking advantage of a unique in vitro assay measuring Hsp90-dependent p53 binding to the promoter sequence. E42A and D88N Hsp90β variants bind but do not hydrolyze ATP, whereas E42A has increased and D88N decreased ATP affinity, compared with WT Hsp90β. Nevertheless, both of these mutants interact with WT p53 with a similar affinity. Surprisingly, in the case of WT, but also E42A Hsp90β, the presence of ATP stimulates dissociation of Hsp90-p53 complexes and results in p53 binding to the promoter sequence. D88N Hsp90β is not efficient in both of these reactions. Using a trap version of the chaperonin GroEL, which irreversibly captures unfolded proteins, we show that Hsp90 chaperone action on WT p53 results in a partial unfolding of the substrate. The ATP-dependent dissociation of p53-Hsp90 complex allows further folding of p53 protein to an active conformation, able to bind to the promoter sequence. Furthermore, in support of these results, the overproduction of WT or E42A Hsp90β stimulates transcription from the WAF1 gene promoter in H1299 cells. Altogether, our research indicates that ATP binding to Hsp90β is a sufficient step for effective WT p53 client protein chaperoning.

MicroRNAs (miRNAs) are a recently discovered category of small RNA molecules that regulate gene expression at the post-transcriptional level. Accumulating evidence indicates that miRNAs are aberrantly expressed in a variety of human cancers and revealed to be oncogenic and to play a pivotal role in initiation and progression of these pathologies. It is now clear that the inhibition of oncogenic miRNAs, defined as blocking their biosynthesis or their function, could find an application in the therapy of different types of cancer in which these miRNAs are implicated. Here we report the design, synthesis, and biological evaluation of new small-molecule RNA ligands targeting the production of oncogenic microRNAs. In this work we focused our attention on miR-372 and miR-373 that are implicated in the tumorigenesis of different types of cancer such as gastric cancer. These two oncogenic miRNAs are overexpressed in gastric cancer cells starting from their precursors pre-miR-372 and pre-miR-373, two stem-loop structured RNAs that lead to mature miRNAs after cleavage by the enzyme Dicer. The small molecules described herein consist of the conjugation of two RNA binding motives, i.e., the aminoglycoside neomycin and different natural and artificial nucleobases, in order to obtain RNA ligands with increased affinity and selectivity compared to that of parent compounds. After the synthesis of this new series of RNA ligands, we demonstrated that they are able to inhibit the production of the oncogenic miRNA-372 and -373 by binding their pre-miRNAs and inhibiting the processing by Dicer. Moreover, we proved that some of these compounds bear anti-proliferative activity toward gastric cancer cells and that this activity is likely linked to a decrease in the production of targeted miRNAs. To date, only few examples of small molecules targeting oncogenic miRNAs have been reported, and such inhibitors could be extremely useful for the development of new anticancer therapeutic strategies as well as useful biochemical tools for the study of miRNAs' pathways and mechanisms. Furthermore, this is the first time that a design based on current knowledge about RNA targeting is proposed in order to target miRNAs' production with small molecules.

Liver cancer, especially hepatocellular carcinoma (HCC), is an aggressive disease with an extremely high mortality rate. Unfortunately, no promising markers are currently available for the early diagnosis of this disease. Thus, a reliable biomarker reflecting the early behaviour of the tumour will be valuable for diagnosis and treatment. The Ras homologous (Rho) GTPases, which belong to the small guanosine triphosphate (GTP) binding proteins, have been reported to play an important role in mediating liver cancer based on their important function in cytoskeletal reorganisation. These proteins can be either oncogenic or tumour suppressors. They are also associated with the acquirement of malignant features by cancer cells. The overexpression of RhoA and Rac1, members of the Rho GTPases, have been linked with carcinogenesis and the progression of different types of cancer. In the quest of elucidating the role of mechanical stimulation in the mechanobiology of liver cancer cells, this paper evaluates the effect of stretching on the expression levels of RhoA and Rac1 in different types of liver cancers. It is shown that that stretching liver cancer cells significantly increases the expression levels of RhoA and Rac1 in HCC and cholangiocarcinoma cell lines. We hypothesise that this relatively simple and sensitive method could be helpful for screening biological features and provide suitable treatment guidance for liver cancer patients.