[Impact of СD8/FoxP3 T lymphocyte ratio in the peritumoral area of primary cutaneous melanoma on the prognosis of the disease]. / Vliyanie sootnosheniya CD8/FoxP3 T-limfotsitov v peritumoral'noi zone pervichnoi melanomy kozhi na prognoz techeniya zabolevaniya.

UNLABELLED: Cutaneous melanoma (CM) is a malignant tumor characterized by typical histological features, one of which is tumor-infiltrating lymphocytes (TIL) that reflect the state of local immunity and determines the course of the disease. AIM: to study a correlation of the ratio of CD8/Foxp3 T lymphocyte subpopulations infiltrating primary cutaneous melanoma (PCM) with the clinical and morphological factors and prognosis of the disease. MATERIAL AND METHODS: The CD8+ and FoxP3+ T-lymphocyte subpopulations infiltrating PCM were investigated in 180 cases by immunohistochemical staining with anti-CD8 and anti-FoxP3 antibodies. RESULTS: The predominant type of TIL was CD8+ cytotoxic T lymphocytes (80.1 cells in the field of vision); FoxP3+ T lymphocytes averaged 34.9 cells in the field of vision. There was a statistically significant correlation of a high ratio of CD8/FoxP3 T cells with disease stage, ulceration and regression segments in PCM, a low disease progression rate, and higher 5-year overall and relapse-free survival. CONCLUSION: The findings may be used in researches and in the practical work of pathologists and clinicians.

Novel TP53 gene mutations in tumors of Russian patients with breast cancer detected using a new solid phase chemical cleavage of mismatch method and identified by sequencing.

Mutations in the tumor-suppressor p53 gene TP53 are frequent in most human cancers including breast cancer. A new solid phase chemical cleavage of mismatch method (CCM) allowed rapid and efficient screening and analysis of the TP53 gene in DNA samples extracted from tumors of 89 breast cancer patients. The novel CCM technique utilized silica beads and the potassium permanganate/tetraethylammonium chloride (KMnO(4)/TEAC) and hydroxylamine (NH(2)OH) reactions were performed sequentially in a single tube. Mutation analysis involved amplification of five different fragments of the TP53 gene using DNA from the 89 tumor samples, then pairing of the 391 labeled PCR products and forming heteroduplexes. A total of 41 unique signals were revealed in the analysis of TP53 exons 5-9 and eight were identified by direct sequencing. The three novel mutations detected are c.600T>G (p.Asn200Lys), c.601T>G (p.Leu201Val), and c.766-768delACA (p.Thr256del). The detected mutations c.638G>T (p.Arg213Leu), c.730G>T (p.Gly244Cys), and c.758C>T (p.Thr253Ile) have not been reported in breast cancer but have been recorded in tumors of other organs. A previously reported mutation c.535C>T (p.His179Tyr) and a heterozygous polymorphism c.639A>G were also detected. Of the 41 unique signals, 36 were not identified as a sequence change. As direct sequencing requires the mutant allele concentration to be greater than 30% when the mutant allele is present in a mixture with the wild-type allele, the CCM method represents a more sensitive technique requiring a lower mutant allele concentration in the wild-type mixture compared with direct sequencing. This reveals the advantage of CCM for unknown point mutation detection in DNA samples of cancer patients.