Genetic and genomic analysis modeling of germline c-MYC overexpression and cancer susceptibility.

BACKGROUND: Germline genetic variation is associated with the differential expression of many human genes. The phenotypic effects of this type of variation may be important when considering susceptibility to common genetic diseases. Three regions at 8q24 have recently been identified to independently confer risk of prostate cancer. Variation at 8q24 has also recently been associated with risk of breast and colorectal cancer. However, none of the risk variants map at or relatively close to known genes, with c-MYC mapping a few hundred kilobases distally. RESULTS: This study identifies cis-regulators of germline c-MYC expression in immortalized lymphocytes of HapMap individuals. Quantitative analysis of c-MYC expression in normal prostate tissues suggests an association between overexpression and variants in Region 1 of prostate cancer risk. Somatic c-MYC overexpression correlates with prostate cancer progression and more aggressive tumor forms, which was also a pathological variable associated with Region 1. Expression profiling analysis and modeling of transcriptional regulatory networks predicts a functional association between MYC and the prostate tumor suppressor KLF6. Analysis of MYC/Myc-driven cell transformation and tumorigenesis substantiates a model in which MYC overexpression promotes transformation by down-regulating KLF6. In this model, a feedback loop through E-cadherin down-regulation causes further transactivation of c-MYC. CONCLUSION: This study proposes that variation at putative 8q24 cis-regulator(s) of transcription can significantly alter germline c-MYC expression levels and, thus, contribute to prostate cancer susceptibility by down-regulating the prostate tumor suppressor KLF6 gene.

Identification of somatic and germline mitochondrial DNA sequence variants in prostate cancer patients.

Prostate cancer (PCa) is the second most frequent cancer among men in the European Union and the most common in the United States. Older age and a positive family history of PCa are important risk factors, but little is known about the disease aetiology. Mitochondria are involved in essential cellular pathways, some of which have been associated with tumorigenesis. We analysed the presence of sequence variants, depletion and rearrangements in the mitochondrial DNA (mtDNA) of PCa patients. Sequencing of the D-loop and genes RNR1 and 2, ND3, ND4L and ND4, and long-range and real-time PCR techniques were carried out on 51 samples (41 from patients and 10 from controls). Normal, hyperplastic or tumour samples were obtained from 17 patients. Six patients' seminal vesicles were also investigated as an additional patient's control tissue (these structures seldom develop tumours). Neither depletion nor mtDNA rearrangements were detected. In contrast, 94 mtDNA sequence variants were identified, 9 previously unreported. The regions presenting more sequence variants were MT-DLOOP (52%), MT-RNR2 (14%) and MT-ND4 (13%). The patients' seminal vesicles studied showed the same set of variants as the corresponding prostate, suggesting either that the pathogenic role of these particular variants is minor or that they participate in the prostatic carcinogenesis in combination with other factors absent in seminal vesicles. Five patients (29.4%) harboured eight somatic changes in the mtDNA. One affects a conserved residue and three have not been previously described. The analysis of other genes in the mtDNA molecule might demonstrate an even higher incidence of mtDNA somatic variants in these PCa patients.

Tissue imprints or primary cultures: which strategy to use to study cytogenetic clonality?

Touch preparations or imprints have been extensively used in cytogenetics to avoid primary cultures, especially when studying solid tumors which are hard to grow in vitro. Interphase nuclei studies by FISH have been validated in several sample types; however, to our knowledge, a comparison between both methods when studying clonality has not yet been published. We have performed a comparative FISH study between touch preparations and cultured cells to assess their reliability when studying the aneuploidy of chromosome Y in mosaicism. Our results in 23 samples indicate that aneuploidy of chromosome Y assessed in cells from tissue cultures versus cells obtained from touch preparations from seminal vesicles of patients with prostate cancer is not comparable. The percentage of aneuploid cells is higher in cultured cells. Attention, therefore, must be paid not to overestimating or underestimating the number of aneuploid cells detected when using interphase FISH studies, especially in solid tumors where clonality is very frequent. Also, according to our results, it is reasonable to extrapolate that when performing interphase nuclei studies in paraffin sections or tissue microarray, and therefore underestimations of aneuploidy could be reported. This might be of special relevance if the aneuploidy detected correlates with the tumor progression or might be used as a prognostic factor.

Altered expression of 12S/MT-RNR1, MT-CO2/COX2, and MT-ATP6 mitochondrial genes in prostate cancer.

BACKGROUND: Prostate cancer is one of the commonest cancers worldwide and is responsible for nearly 6% of all male cancer deaths. Despite this relevance, the mechanisms involved in the development and progression of this malignancy remain unknown. The involvement of polypeptides of the mitochondrial respiratory chain, the Krebs cycle and the glutathione antioxidant system in this type of cancer has been previously described, although no publication has focused on the expression of mitochondrial genes in the prostate of PCa patients. METHODS: We have determined by reverse transcription-quantitative PCR (RT-qPCR) the relative amount of the transcripts of eight mitochondrial genes (MT-ND2, MT-ND4, MT-ND6, MT-CYB, 12S/MT-RNR1, 16S/MT-RNR2, MT-CO2/COX2, MT-ATP6), and four nuclear genes (COX11, GSR, CS, ACO2), all of them key players in the normal metabolism of mitochondria. Additionally we analyzed the expression of Cyclophilin A (PPIA). RESULTS: We observed differential expression of mitochondrial 12S/MT-RNR1, MT-CO2/COX2, and MT-ATP6 transcripts in tumor samples when compared to their paired normal samples. CONCLUSIONS: The amount of mitochondrial 12S/MT-RNR1, MT-CO2/COX2, and MT-ATP6 transcripts is significantly decreased in tumor samples when compared to their paired normal sample, suggesting that mitochondrial gene expression is altered in PCa.

Aneuploidy of chromosome Y in prostate tumors and seminal vesicles: a possible sign of aging rather than an indicator of carcinogenesis?

Chromosome Y aneuploidies have been reported as one of the recurrent cytogenetic findings in prostate cancer (PCa) and many other solid and hematological tumors. We have studied this aneuploidy in 28 patients with PCa undergoing radical prostatectomy, one patient with benign hyperplasia (BPH) and four organ donors. A total of 72 samples have been studied: 17 tumors, 25 nontumor prostate tissues, 1 BPH, 21 seminal vesicles samples obtained along with the prostate when patients underwent radical prostatectomy and prostate tissues and seminal vesicles from four organ donors. We have also studied the aneuploidy of chromosome Y in peripheral blood from four of the patients and in seminal vesicles of 11 individuals with bladder cancer (BC). The study has been performed by Fluorescence in situ hybridization (FISH) in uncultured cells. Our results indicate that complete loss of chromosome Y is found in almost all the seminal vesicles both from patients with PCa and patients with BC (samples obtained from the tissue bank), and is more frequent in prostate tumors than in nontumor samples. The percentages of chromosome Y loss in the tissues analyzed are significatively higher than expected in lymphocytes considering the patient's age as reported in the literature. The high percentage of chromosome Y loss found in the nonmalignant seminal vesicles of these patients may be an indicator of an ageing process rather than a primary cytogenetic alteration in the carcinogenesis of the prostate. However, a contribution of this loss to chromosomal instability and therefore, to the multistep tumorigenic process, cannot be discarded.

[Diagnostic methodology for the biochemical recurrence of prostate cancer after brachytherapy]. / Metodología diagnóstica ante la recidiva bioquímica después de braquiterapia.

Currently prostate cancer is the most frequent extracutaneous neoplasia in males in the USA, and second after lung cancer in our country. Over the last years the profile of prostate cancers diagnosed has changed due to the wide diffusion of PSA determination. Currently, almost 47% of prostate cancers are low risk at diagnosis. In this situation, the minimally invasive therapies such as brachytherapy have a growing acceptance in our environment. We analyze the special PSA kinetics after brachytherapy, and the difficulty entailed by the diagnosis of biochemical recurrence after brachytherapy, performing a bibliographic review of the available scientific evidence.