[Primary multiple malignant lung tumors. References and a clinical observation].

The problem of primary multiple tumors is relevant to current clinical oncology because of increasing of number of patients with multiple malignant tumors and unsolved issues of treatment. Primary multiple malignant lung tumors is a common oncological situation requires an individualized, differentiated approach to treatment. The results of treatment are associated with the prevalence of the process, stages of tumor development, spare capacity of patients. There is presented clinical example of a patient with metachronous primary multiple malignant tumors of one lung.

NON-INVASIVE MONITORING OF EGFR MUTATIONS IN NONSMALL CELL LUNG CANCER (NSCLC) DURING TARGETED THERAPY: STATE OF THE ART AND OUR EXPERIENCE.

Liquid biopsy is a promising approach to molecular tumor testing in the context of targeted therapy. During this pilot study we applied a high-sensitivity protocol for detection of tumor-derived mutations in circulating plasma DNA of EGFR-positive non-small cell lung cancer (NSCLC) patients during EGFR-TKI therapy. We showed that this protocol was well suited for dynamic monitoring during targeted therapy as well as for detection of acquired resistance mutations.

[Radical removal of a malignant mediastinum hemangioendothelioma with restoration of the main blood supply to the brain].

46 year old man appealed to the Cancer Research Center of RAMS in October 2012 with unverified anterior superior mediastinal tumor, which was diagnosed in 2010. Progressive compartment syndrome of the superior vena cava was observed. On examination: CT, MRI, angiography, histological and cytological examination of biopsy material did not allow to confirm the morphological structure of the tumor. Removal of the tumor with bifurcation of the brachiocephalic trunk prosthetics was performed. Immunohistochemical (IHC) study verified malignant hemangioendothelioma.

[Arf6, RalA and BIRC5 protein expression in non small cell lung cancer].

Evaluation of tumor markers expression pattern which determines individual progression parameters is one of the major topics in molecular oncopathology research. This work presents research on expression analysis of several Ras-Ral associated signal transduction pathway proteins (Arf6, RalA and BIRC5) in accordance with clinical criteria in non small cell lung cancer patients. Using Western-blot analysis and RT-PCR Arf6, RalA and BIRC5 expression has been analyzed in parallel in 53 non small cell lung cancer samples of different origin. Arf6 protein expression was elevated in 55% non small cell lung cancer tumor samples in comparison with normal tissue. In the group of squamous cell lung cancer Arf6 expression elevation was observed more often. RalA protein expression was decreased in comparison to normal tissue samples in 64% of non small cell lung cancer regardless to morphological structure. Correlation between RalA protein expression decrease and absence of regional metastases was revealed for squamous cell lung cancer. BIRC5 protein expression in tumor samples versus corresponding normal tissue was 1.3 times more often elevated in the squamous cell lung cancer group (in 76% tumor samples). At the same time elevation of BIRC5 expression was fixed only in 63% of adenocarcinoma tumor samples. A statistically significant decrease (p = 0.0158) of RalA protein expression and increase (p = 0.0498) of Arf6 protein expression in comparison with normal tissue was found for T1-2N0M0 and T1-2N1-2M0 groups of squamous cell lung cancer correspondingly.

[Significance of the expression of nucleolar argyrophilic proteins and antigen Ki-67 in the evaluation of cell proliferative activity and in the prediction of minimal (T1) lung cancer].

S u m m a ry. - The subject of the study was 20 cases of non-small-cell lung carcinomas, up to 3 cm in diameter, conventionally designed as minimal lung cancers removed in patients operated on at the N. N. Blokhin Cancer Research Centre, Russian Academy of Medical Sciences in 1986 to 2001. According to survival rates after surgery, the patients were divided into two groups: 1) those who died within the first two years; 2) those who were followed up for 3-5 years. Histological, histochemical, and immunohistochemical studies were performed. The expression of argyrophylic nucleolar organizer site proteins (Ag-NOS-proteins) that characterized the rate of cell proliferation (the duration of a cellular cycle) and the expression of Ki-67 antigen, which reflected the fraction of growth (the number of proliferating cells), were revealed in the tumor cells. Minimal lung cancers were found to be a heterogeneous group of neoplasms showing differences in both the rate of cell proliferation and the count of proliferating cells. The cell proliferation rate is a determinant of the clinical course of minimal lung cancers. Group 1 tumors characterized by the superexpression of Ag-NOS-proteins and, accordingly, the higher cell proliferation rate and the moderate count of proliferating cells had a poor prognosis even in the presence of Stage IA whereas Group 2 tumors with a large quantity of proliferating cells, but with the less rate of cell proliferation were characterized by a much better prognosis. The rate of cell proliferation (expression of Ag-NOS-proteins) and the count of proliferating cells (the expression of Ki-67 antigen) should be simultaneously studied to have more complete information on the proliferative potential of tumor cells and on the prediction of the course of neoplasms.

[Transcription TIMP3, DAPk1 and AKR1B10 genes in squamous cell lung cancer].

Lung cancer is one of the most frequent neoplasia in the Russia, the United States and Europe. This cancer is associated with functional activity changes of many genes. In the present study TIMP3, DAPK1 and AKR1B10 genes transcription analysis of squamous cell lung cancer specimens was carried out using reverse transcription-PCR. Substantial increasing of AKR1B10 transcription level is revealed in 80% tumor samples. TIMP3 and DAPK1 transcription level is considerably decreased in 76 and 72% tumor specimens, accordingly. These results may point out that all three genes are important for squamous cell lung cancer tumorogenesis while AKR1B10 is potential oncogene whereas TIMP3 and DAPK1 are potential tumor suppressor genes. We suggest that revealed substantial transcription level-changes of investigated genes may be used for oncodiagnostics.

[Inhibition of ABC-transporter(s)' function in non-small cell lung cancer cells by platinum drugs]. / Ingibirovanie funktsii ABC-transporterov v kletkakh nemelkokletochnogo raka legkogo pri vozdeistvii preparatov platiny.

With an account of the literature data that platinum drugs react with many cellular targets, including ATP and proteins, the authors suggested that disturbance of the function of energy-dependent ABC-transporters (markers of multidrug resistance, MDR) under the effect of platinum drugs could be a cause of increased efficacy of MDR agents (agents, MDR to which is developed by the classical mechanism) when used in combination with platinum drugs even in the treatment of multidrug resistant lung cancer. The cisplatin and carboplatin effect on accumulation of MDR doxorubicin in cells of non-small cell cancer was studied by flow cytometry with the use of biopsy specimens. The MDR phenotype of the tumors was determined by a change in doxorubicin intracellular accumulation under the action of the ABC-transporter(s)' inhibitors: verapamil and genistein (specific inhibitors of Pgp and MRP respectively) and sodium azide (an inhibitor of all energy-dependent ABC-transporters). The MDR phenotypes, i.e. Pgp-MRP+ or Pgp+MRP+, were detected in all the tumors investigated. Two types of changes in doxorubicin intracellular accumulation under the action of the inhibitors and the platinum drugs were shown: (a) an increase in doxorubicin cytoplasmic accumulation and (b) a change in subcellular distribution of the anthracycline (increased accumulation of doxorubicin in the cell nucleus and its higher binding to DNA). Cisplatin and carboplatin had an inhibitory effect on ABC-transporter(s) in all the tumors investigated but the effect of carboplatin was less pronounced. It was concluded that cisplatin and carboplatin stimulation of doxorubicin intracellular accumulation, as well as a change in subcellular distribution of the anthracycline under the action of the platinum drugs (increased doxorubicin accumulation in the cell nucleus) in multidrug resistant lung tumors could be at least partly explained by inhibition of the MDR transporter(s)' function. The results could provide a basis for the use of the sequential combination cisplatin (or carboplatin)-->doxorubicin in the treatment of multidrug resistant lung cancer.

[Comparative assessment of the human cell viability ex vivo in Hanks solutions in distilled and Penta water]. / Sravnitel'naia otsenka zhiznesposobnosti kletok cheloveka ex vivo v rastvore Khenksa na distillirovannoi vode i vode Renta.

Assessment of human colon and lung mucosa cell viability was performed in Hanks salt media prepared separately with distilled and patented Penta water. The cell viability in the suspension was estimated by fluorescence intensity of propidium iodide, a DNA specific dye, that is an indicator of DNA structure intactness or damage. The experiments were conducted with the flow cytometry technique. The histogram analysis showed that 2-hour incubation of the cells in Hanks salt medium prepared with distilled water resulted in an increase of the number of the apoptotic cells with a respective decrease of the number of the intact cells (approximately 2- and 4-fold in the suspensions of the colon and lung mucosa cells respectively). A similar experiment with Hanks salt medium prepared with Penta water resulted in a less marked increase of the viability of the apoptotic cells that did not exceed 20 and 50% for the colon and lung mucosa cells respectively. The findings showed that viability of the cells ex vivo was significantly higher when Penta water was used as a solvent for preparing Hanks salt media as compared to distilled water. The result is important for ex vivo experiments since maximum preservation of the DNA structure minimizes the number of possible experimental inaccurate and consequently erroneous conclusions. Furthermore, the fact of pathologic process inhibition in cells isolated from various human tissues in Penta-based salt media is in favour of using Penta water as a solvent for nutritional ingredients in ex vivo maintenance of human tissues for transplantation as compared to distilled water.

[Profile of methylation of certain tumor growth suppressing genes in non-small cell lung cancer]. / Profil' metilirovaniia nekotorykh genov-supressorov opukholevogo rosta pri nemelkokletochnom rake legkogo.

Multiplex methylation-sensitive PCR was employed in studying the methylation of CpG islands in the RB1, p16/CDKN2A, p15/CDKN2B, p14/ARF, CDH1, HIC1, and N33 5' regions in non-small cell lung cancer (51 tumors). Methylation was observed for the two suppressor genes involved in controlling the cell cycle through the Cdk-Rb-E2F signaling pathway, RB1 (10/51, 19%) and p16 (20/51, 39%). The highest methylation frequencies were established for CDH1 (72%) and HIC1 (82%). The CpG islands of p14 and p15 proved to be nonmethylated. At least one gene was methylated in 90% (46/51) tumors and no gene, in 10% (5/51) tumors. In addition, the genes were tested for methylation in peripheral blood lymphocytes of healthy subjects. Methylation frequency significantly differed between tumors and normal cells in the case of RB1, p16, CDH1, HIC1, and N33. Gene methylation frequency was tested for association with histological type of the tumor and stage of tumor progression. Methylation index of a panel of tumor suppressor genes was established for groups of tumors varying in clinical and morphological parameters.

Expression of phosphatidylinositol-3 kinase in lung cancer.

The expression of phosphatidylinositol-3 kinase in tumors and homologous tissues from 29 patients with lung cancer, 5 patients with lung metastases of various tumors, and some non-tumorous pulmonary diseases was studied by Western blot analysis. The expression of phosphatidylinositol-3 kinase was increased in these tumors in comparison with histologically intact lung tissue in 5 patients with non-small-cell cancer. In 20 patients expression of phosphatidylinositol-3 kinase was the same as in homologous tissue and in 4 patients it was decreased. No relationship between phosphatidylinositol-3 kinase expression and clinical and morphological characteristics of lung cancer was revealed.

Allelic imbalance and instability of microsatellite loci on chromosome 1p in human non-small-cell lung cancer.

The mapping of allelic loss on the short arm of chromosome 1 has been performed in non-small-cell lung cancer. We used a set of 11 microsatellite loci spanning 1p to examine the frequency of allelic imbalance in a panel of 58 tumours. Fifty-one of 58 (87.9%) cases have shown somatic allelic loss at one or more loci tested. The two shortest regions of the overlap (SRO) of the deletions have been identified: SRO 1 at 1p13.1 and SRO 2 at 1p32-pter. Allelic losses at these regions have been compared among adenocarcinoma and squamous cell carcinoma and no difference has been found. In contrast to SRO 1, deletions at SRO 2 significantly correlated with advanced stage of the disease as well as post-operative metastasizing and relapse. These data may suggest that SRO 1 and SRO 2 can harbour tumour-supressor genes (TSGs) involved in different stages of NSCLC development. SRO 2 is still quite large and its refined mapping should help attempts to clone and identify the putative TSG(s). Microsatellite instability (replication errors) affecting only 6 (10.3%) of 58 tumour samples is an infrequent genetic alteration at the loci tested.