Molecular Characteristics of Bladder Tumor: Increased Gene Expression of MAGE-A6 and MAGE-A11 with Decreased MicroRNA-34a and MicroRNA-125b.

Bladder cancer is recognized as one of the top ten most common cancers worldwide. Activation of oncogenes, inactivation of tumor suppressor genes, and dysregulation of androgen signaling pathways are three major pathophysiological causes in the development of bladder tumors. Discovering potential biomarkers is required for the management and immunotherapy of bladder cancer. Melanoma-associated antigen (MAGE)-A6 and MAGE-A11 are two cancer-testis antigens that are potential coregulators of androgen receptors. MicroRNAs, especially miR-34a and miR-125b are two important tumor suppressors that play a critical role in regulating different signaling pathways and inhibiting tumor development. Twenty-nine surgical tissue biopsies were collected from patients with no preoperative chemotherapy or radiotherapy (26 males and, 3 females, mean age±SD: 62.4±13.3 years). Seventeen adjacent uninvolved tissues with no abnormalities upon histological examination were considered normal controls (14 males and, 3 females, mean age±SD: 64.2±7.4 years) . Quantitative PCR was performed to evaluate the gene expression level of MAGE-A6, MAGE-A11, miR-34a, and miR-125b in bladder cancer biopsies. MAGE-A6 and MAGE-A11 expressions were significantly increased in bladder tumors compared with normal tissues. However, the expression levels of miR-34a and miR-125b were significantly downregulated in bladder tumor tissues. Interestingly, the expression level of all these genes was significantly associated with tumor grade, pathological stage (pT), and muscular invasion. MAGE-A6 and MAGE-A11 can be considered potential markers for the diagnosis and immunotherapy of bladder tumors. Furthermore, the modulation of miR-34a and miR-125b gene expression in association with increased MAGE-A6 and MAGE-A11 genes could open a new horizon in the improvement of bladder cancer.

Overexpression of long intergenic noncoding RNAs in bladder cancer: A new insight to cancer diagnosis.

There is increasing evidence that show long noncoding RNAs including long intergenic noncoding RNAs (lincRNA) play a pivotal regulatory role in the biological processes. Differential expression of lincRNAs can be utilized for cancer diagnosis, prognosis, and targeted therapy. Little is known about their expressions in urothelial tumors. Concerning the potential role of lincRNAs in cancer development, we aimed to investigate the expression levels of LINC00958 and DNM3OS in bladder cancer. Fifty tumor and 50 adjacent non-tumor tissue samples along with their clinicopathological parameters were obtained from bladder cancer patients. Expressions of LINC00958 and DNM3OS were analyzed by Real-time PCR. ROC curve analysis was used to evaluate the diagnostic power of LINC0095 and DNM3OS for BC. Expression level of LINC00958 was considerably increased in cancerous tissues (P < 0.001) and in correlation with cigarette smoking (P = 0.043). DNM3OS expression was higher in the tumor tissues than normal tissues (P < 0.001) and showed a significant association with age (P = 0.038). By using the ROC curve, the diagnostic power of LINC00958 and DNM3OS transcript levels in bladder cancer were estimated to be 87% and 75%, respectively. Our findings offer some important intuitions into the oncogenic role of LINC00958 and DNM3OS in bladder cancer and suggest that they can be candidate biomarkers and may provide new approaches for the diagnosis and therapy if being validated in a larger sample size of clinical samples as well as functional studies.

High Resolution Melting Analysis for Rapid Detection of PIK3CA Gene Mutations in Bladder Cancer: A Mutated Target for Cancer Therapy.

PURPOSE: PIK3CA gene mutations have clinical importance and their presence is associated with therapy response. They are also considered as a molecule for targeted therapy. As regards to their importance, genetic variation within a population as well as among different populations, this study was conducted to detect common mutations of exons 9 and 20 and other probable mutations in PIK3CA gene as well as their frequencies in Iranian bladder cancer patients. MATERIALS AND METHODS: Paired tumor and adjacent normal tissues samples were obtained from 50 bladder cancer patients. Mutations of PIK3CA gene were detected using High Resolution Melting (HRM) analysis which is ahighly sensitive, repeatable, rapid, and cost-effective technique. To determine the precision of the HRM analysis, Sanger sequencing analysis was used. RESULTS: The result showed that mutations were present in 10% (5/50) of the subjects. The majority of these cases (4/5) had the mutation(s) in exon 9, spanning over five different mutations, among which three of them were actually novel mutations. Further analysis showed that 2 cases had simultaneous mutations for exon 9. In addition to novel mutations, the PIK3CA mutation rate observed in Iranian bladder patients was not as frequent as previous reports and COSMIC. CONCLUSION: HRM can be used as a rapid and sensitive method for mutation screening. Dysregulation of PIK3CA gene in bladder cancer reveals its potentials as a mechanistic link for cancer development, which in turn suggests its special use in interventional studies for targeted therapy.

Dysregulated Expression of Long Intergenic Non-coding RNAs (LincRNAs) in Urothelial Bladder Carcinoma.

Long intergenic non-coding RNA (lincRNA) has been introduced as key regulators of diverse biological processes, including transcription, chromatin organization, cell growth and tumorigenesis. With regard to the potential role of lincRNAs in cancer development, one may postulate that differential expression of lincRNAs could be employed as a tool in cancer diagnosis, prognosis, and targeted therapy. In this study, we aimed to explore the putative correlation between the expression levels of two lincRNAs: LINC00152 and LINC01082 in the bladder cancer (BC), in comparison with its adjacent non-cancerous tissue. Fifty Iranian subjects diagnosed with BC, representing in different stages and grades participated in this study The mRNA expression levels of the abovementioned lincRNAs were comparatively analyzed in cancerous and their adjacent non-cancerous counterpart tissues, of each subject by Real-Time PCR. The expression levels of LINC00152, and LINC01082 were significantly lower in tumor tissues in comparison with their adjacent normal tissues (P<0.001). More notably, in the case of LINC01082 the reduced expression was differentiated by the muscle invasiveness pattern of the tumor (P= 0.05). Our study presents a new finding about the tumor suppressor potentiality of these lincRNAs in BC development that in turn may suggest them as candidate biomarkers. Replicating this study in higher number of BC subjects, coupled with functional analysis, is necessary to investigate interconnections between these RNAs and cancer development, leading to better understanding of cancer biology.

Expression of two testis-specific genes, SPATA19 and LEMD1, in prostate cancer.

BACKGROUND AND AIMS: Screening methods for early detection of prostate cancer have some limitations regarding specificity and sensitivity, so there is a continuing search to find new cancer biomarkers. Cancer-testis genes are a group of genes with expression almost limited to testis and different kinds of tumors. Since testis is an immune privileged site, if these genes are expressed in tumors, they can be immunogenic. We undertook this study to find new members of the cancer-testis gene family appropriate for cancer immunotherapy METHODS: We analyzed the expression of six testis-specific genes called ODF1, ODF2, ODF3, ODF4, LEMD1 and SPATA19 in 30 prostate cancer and 25 benign prostate hyperplasia (BPH) samples by RT-PCR and restriction fragment length polymorphism (RFLP). RESULTS: Of the prostate cancer samples, 10, 10, 23 and 40% showed ODF1, ODF2, LEMD1 and SPATA19 specific bands, respectively, but none of the BPH samples expressed any of these genes. The difference between prostate cancer and BPH groups for LEMD1 and SPATA19 expression was significant. Mean serum PSA level was significantly higher in patients expressing ODF2 than in the other patients. CONCLUSIONS: ODF1, ODF2, SPATA19 and LEMD1 are members of cancer-testis gene family. In addition, LEMD1 and SPATA19 are putative cancer biomarkers and promising targets for active immunotherapy.

Molecular analysis of the SMN1 and NAIP genes in Iranian patients with spinal muscular atrophy.

INTRODUCTION: Childhood-onset proximal spinal muscular atrophies (SMAs) are an autosomal recessive, clinically heterogeneous group of neuropathies characterised by the selective degeneration of anterior horn cells. SMA has an estimated incidence of 1 in 10,000 live births. The causative genes are survival motor neuron (SMN) gene and neuronal apoptosis inhibitory protein (NAIP) gene. Deletions of the telomeric copy of SMN gene (SMN1) have been reported in 88.5% to 95% of SMA cases, whereas the deletion rate for NAIP gene (NAIP) is between 20% and 50% depending on the disease severity. The main objective of this study was to genetically characterise the childhood onset of SMA in Iran. MATERIALS AND METHODS: Molecular analysis was performed on a total of 75 patients with a clinical diagnosis of SMA. In addition to common PCR analysis for SMN1 exons 7 and 8, we analysed NAIP exons 4 and 5, along with exon 13, as a internal control, by bi-plex PCR. RESULTS: The homozygous-deletion frequency rate for the telomeric copy of SMN exons 7 and 8 in all types of SMA was 97%. Moreover, exons 5 and 6 of NAIP gene were deleted in approximately 83% of all SMA types. Three deletion haplotypes were constructed by using SMN and NAIP genotypes. Haplotype A, in which both genes are deleted, was seen in approximately 83% of SMA types I and II but not type III. It was also found predominantly in phenotypically severe group with an early age of onset (i.e., less than 6-month-old). We also report 34 of our prenatal diagnosis. CONCLUSIONS: To our knowledge, the present study is the first one giving detailed information on SMN and NAIP deletion rates in Iranian SMA patients. Our results show that the frequency of SMN1 homozygous deletions in Iran is in agreement with previous studies in other countries. The molecular analysis of SMA-related gene deletion/s will be a useful tool for pre- and postnatal diagnostic.