Changes in the genetic landscape during the malignization of high grade squamous intraepithelial lesion into cervical cancer.

In 5 patients, a change in the genetic landscape from HPV16 positive high-grade squamous intraepithelial lesion (HSIL) to squamous cervical cancer was traced, which occurred in these patients within the period from 7 months to 5 years after diagnosing HSIL. MATERIALS AND METHODS: The DNA from paraffin blocks of dysplasia tissue and the tumor that emerged afterwards was used for the study, which was analyzed using the OncoScan FFPE microarray Assay Kit Affymetrix (USA) for genome-wide determination of gene abundance and 65 key somatic driver mutations of oncogenes and tumor suppressor genes. RESULTS: In the study of HSIL material, somatic mutations were observed in 4/5 cases, 18 different somatic driver mutations of the NRAS, EGFR, BRAF, KRAS, IDH2 oncogenes and TP53 suppressor genes were found and almost no CNA-Copy Number Aberration was identified. HSIL malignization is associated with the appearance of secondary driver mutations in oncogenes and tumor suppressor genes and a large number of structural and numerical CNA, the frequency of which correlates with the time of dysplasia malignization into cancer with a very high correlation coefficient r = 0.98, P = 0.004. The trees of dysplasia evolution into tumor were constructed for each patient. CONCLUSION: According to the results of the work, it is assumed that the initiation of the development of mucosa dysplastic changes is due to primary driver mutations. The formation of secondary driver mutations and CNA are genetic mechanisms of malignant transformation, while the scenarios of the evolution of dysplasia into a tumor are individual and very diverse.

[Proteins of insuline-like growth factor system in tumor and ascites in ovarian cancer: the relation with neoajuvant chemotherapy effectiveness].

The purpose of the study was to investigate insulin-like growth factors (IGFs) levels in primary tumors and ascites be- fore neoadjuvant chemotherapy (NACT) for searching markers -predictors, associated with its efficiency. The content of IGFs, IGFBPs and receptor IGF-IR is analyzed in primary tumor tis- sues and ascites of 47 patients with high grade ovarian serous adenocarcinomas. The results show a high concentration of IGFs and IGFBPs in ascites as compared with tumor samples. It is found the increase of IGF-II as well as the decrease of IGFBP-3 and tyrosine kinase receptor IGF-IR levels in I stage cancer patients in comparison to patients with advanced ovar- ian cancer. It is obtained the fact of dependence IGFs and IGFBPs levels on the volume of ascites. This factors as well as IGFs protein content are significant markers - predictors of NACT efficiency in group of patients with high grade serous adenocarcinomas.

[Optimization of the extent of surgery in organ- preserving treatment for invasive cervical cancer (the role of sentinel lymph nodes study)].

The study included 44 patients with Stage Ia1 - lb1 cervi- cal cancer who underwent organ-preserving surgery (transab- dominal trachelectomy). To visualize sentinel lymph nodes - lymphoscintigraphy with injection of radioactive lymphotropic isotope - (99m)Tc-labelled nanocolloid was injected a day before surgery. Intraoperative identification of sentinel lymph nodes using gamma probe was carried out to assess, which lymph nodes had taken up the radionuclide. Detection of sentinel lymph nodes in cervical cancer patients could accurately predict the pelvic lymph node status, assess the stage of the dis- ease, individualize the extent of surgery and determine indica- tions for organ-preserving surgery.

Integrative and episomal forms of genotype 16 of human papillomavirus in patients with cervical intraepithelial neoplasia and cervical cancer.

The study involved 500 patients with LSIL (low grade squamous intraepithelial lesion), HSIL (high grade squamous intraepithelial lesion), stage I-IV cervical cancer, infected with human papillomavirus (HPV), as well as 235 women without pathological changes in cervical mucosa. The comprehensive survey included colposcopy, cytological and histological analysis, detection and genotyping of high-risk human papillomavirus. Viral load and physical status of HPV16 DNA was evaluated in cases of mono-infection (n = 148). The prevalence of virus-positive cases among the patients with LSIL/NSIL, cervical cancer patients and healthy women was 69.2%, 76.7% and 51.9%, respectively. An association between the severity of disease and high viral load was revealed. The frequency of integrated DNA was strongly increased in patients with a high viral load. The frequency of episomal forms was either reduced or not detecteable in patients with high viral load as compared to patients with low viral load. It is reasonable to suggest that a high HPV16 viral load may cause an increase in the frequency of integration of virus DNA into the cellular/host genome. This suggests that a high HPV16 viral load may be considered as a risk factor for prognosis of cervical intraepithelial neoplasia and cervical cancer.

[Serum adipokines and their receptors in endometrial and colon cancer patients: relationship with tumor invasion and metastasis].

The aim of the study was to investigate the serum adipokine levels and expression of adipokine receptors (AdipoR1, AdipoR2) in patients with endometrial and colon cancer in relation with the main clinical morphological parameters (tumor invasion, lymph node involvement). The study included 60 endometrial cancer patients with I-II Stage and 31 patients with colon cancer (T2-4N0-2M0). Serum adipokine levels, the level of soluble form of the leptin receptor (sOb-R) and AdipoR1 and AdipoR2 expression were evaluated with ELISA. In endometrial cancer serum leptin and adiponectin levels were associated not only with metabolic disorders but also with cervical invasion. In colon cancer serum leptin level was associated with lymph node involvement. The data obtained showed the potential implication of serum adipokines into tumor invasion and metastasis. In both sites intratumoral levels of AdipoR1 H AdipoR2 were not associated with the presence of metabolic syndrome. The AdipoR1 level was related with myometrial invasion. In colon cancer patients, AdipoR1 and AdipoR2 expressions were associated with lymph node involvement, and AdipoR1 expression was correlated with tumor size. The obtained results demonstrated involvement of adipose tissue hormones (leptin and adiponectin) and adiponectin receptors (AdipoR1 and AdipoR2) in tumor growth, invasion and lymphogenic metastasis.

[Ultrasound semiotics in recurrent ovarian cancer after optimal cytoreductive surgery].

Features of ultrasound picture of morphologically verified recurrence of ovarian cancer in 21 patients are presented, who received combined treatment including cytoreductive surgery in the form of hysterectomy with oophorectomy, resection of the greater omentum and 6 courses of chemotherapy CAP for ovarian cancer stage III (FIGO). In all patients cytoreductive surgery was optimal--without residual tumor. Recurrence of the disease was detected in 12-48 months in 80.9% of the cases. Three variants of recurrence was revealed by ultrasonography: isolated peritoneal dissemination, in 14.2% of the cases, which was mainly detected during the first 12 months; single entities in the projection of the small pelvis (61.9%) and mixed form (local lesions of small pelvis and peritoneal dissemination) in 23.8% of the cases.

[Expression and activity of proteases in metastasis of ovarian cancer].

The total chymotrypsin-like activity of proteasornes, the activity of 20S- and 26S-proteasome pools and calpains, and the expression of metalloproteinase PAPP-A in primary tumors and metastasized tissues were studied in 13 patients with epithelial ovarian cancer. It was shown that initiation of the process of tumor dissemination occurs against the background of active proteolytic processes; A decrease in activity of 26S-proteasomes and total calpain activity and increased expression ofmetalloproteinase PAPP-A in the primary tu mors were found in patients with ascites as compared with patients without ascites. The disease progression after treatment and achieved stabilization were found in patients with decreased activity of intracellular proteases and a high content of PAPP-A in the primary tumors.

[Association of intracellular proteinase activities with the content of locomotor proteins in tissues of primary tumors and metastasis in ovarian cancer].

The ability to active movement in extracellular matrix wherein significant role plays remodeling of the cytoskeleton by actin-binding proteins may influence on the metastatic potential of tumor cells. We studied the expression of actin-binding proteins and ß-catenin in connection with proteasome and calpain functioning in the tissues of primary tumors and metastases of ovarian cancer. The chymotrypsin-like proteasome activity and calpain activity were shown to be significantly higher in ovarian cancer than in normal tissues. Furthermore, the activity of the proteasome and calpain were significantly higher in the peritoneal metastases in comparison with primary tumors. Correlation analysis showed in the primary tumor tissue the presence of a positive relationship between the activity of calpain and chymotrypsin-like proteasome activity (r = 0.82; p = 0.0005), whereas in metastases this connection was not revealed. Contents of p45 Ser ß-catenin and the actin-severing protein gelzolin were decreased in metastases relative to primary tumors. Level of cofilin, functionally similar to gelzolin protein, was significantly higher in metastases compared to primary ovarian tumor tissue. In ovarian cancer significant reduction in the number of the monomer binder protein thymosin-ß4 was observed in primary tumors and metastases as compared to normal tissues, but significant differences between the primary tumor and metastases were not observed. In the tissues of primary tumors negative correlations were observed between the chymotrypsin-like activity of the proteasome and the amount of p45 Ser ß-catenin and protein Arp3, a member of the Arp2/3 complex. In metastasis negative correlation were revealed between the activity of calpain and content Arp3, cofilin, thymosin. The data obtained suggest the existence of different mechanisms of proteolytic regulation of locomotor proteins in primary tumors and metastases in ovarian cancer.

[Chymotripsin-like activity and subunit composition of proteasomes in human cancers].

Activity of the proteasome, polyfunctional enzymatic complex, is known to undergo changes during cancer development. This phenomenon is, probably, caused by the changes in subunit composition of proteasomes. In present work, we studied chymotrypsin-like activity of proteasomes, subunit composition and their association in breast cancer, head and neck squamous cell carcinoma, endometrial cancer, renal cancer, bladder cancer, stomach cancer and colorectal cancer. The increase of proteasome activity was revealed in most cancer tissues compared with adjacent tissues except for the renal cell carcinoma. Changes in proteasome activity in cancer tissues compared with correspondent normal tissues were accompanied by modification of its subunit composition. High proteasome activity was observed in combination with an increased expression of immune subunits and/or proteasome activator PA28, associated with activity of 20S proteasome. In breast cancer, head and neck squamous cell carcinoma, bladder cancer, stomach cancer and colorectal cancer we additionally found higher expression of Rpt6 subunit of 26S proteasome. Correlations between chymotrypsin like proteasome activity and subunit expressions were found in human cancer tissues. In summary, we suggest that proteasome ac- tivation and changes in its subunit composition plays an important role in cancer pathogenesis.

[Specifics of hormonal and energy balance in patients with hyperplasia and endometrial neoplasia with metabolic syndrome in the background].

We conducted a comparative investigation of the hormonal status (LH, FSH, estradiol, progesterone, testosterone, prolactin, SHBG), energy status (leptin, ghrelin, insulin), and carbohydrate and lipid metabolism in patients with endometrial hyperplasia and neoplasia (168 patients) with or without metabolic syndrome in the background. Patients with metabolic syndrome had a high frequency of elevated estrogen (72%), testosterone (65%), insulin (81%), leptin (68%). There was a marked increase in the basal level of luteinizing hormone, prolactin, index, LH/FSH, but decrease in FSH and progesterone. There were significant changes in carbohydrate and lipid metabolism. The possible mechanisms for the contribution of the investigated factors to the development of the pathological processes in the endometrium are presented.

Proteasome activity and their subunit composition in endometrial cancer tissue: correlations with clinical morphological parameters.

The development of endometrial cancer is related to the status of the intracellular proteasome system. Total proteasome activity and pools 26S and 20S activities are higher in tumor tissue than in intact endometrium, and their composition is different. The expression of α1α2α3α5α6α7 is lower in endometrial cancer tissue in comparison with intact endometrium and the content of immune subunits LMP7, LMP2, and PA28ß is increased. Total proteasome activity depends on the disease stage.

[Regulation of insulin-like growth factors and NF-kappaB by proteasome system in endometrial cancer].

Insulin-like growth factors and the nuclear factor kappaB (NF-kappaB) are known to play an important role in pathogenesis of endometrial cancer. However, there is still uncertainty about their proteolytic regulation. We studied the correlation between chymotrypsin-like activity ofproteasomes and IGF-I, IGF-II and NF-kappaB levels in endometrial cancer tissues. The total activity of proteasomes and the 20S and 26S proteasome activities were shown to be significantly higher in malignant tumors than in unaltered endometrium. Negative correlations between the 26S proteasome activity and NF-kappaBp50 level as well as between the 26S and 20S proteasome activities and IGF-I level were found. The data obtained indicate a possible proteasome regulation of growth and transcription factors. As it is considered that the major pool of IGF-I is located in the extracellular space, it is likely that extracellular proteasomes also take part in the regulation of IGF-I content. The positive correlation between IGF-I level and PAPP-A metalloproteinase gives evidence that this proteolytic enzyme is another important regulator of growth factor level, which provides proteolysis of IGF-I binding proteins and increasing IGF-I concentration in tissues. The present data show possibility of proteolytic regulation of growth and nuclear factors that can play an important role in cancer pathogenesis.

[Proteasome activity in tumors of female reproductive system].

Proteasomes (multiproteinase protein complexes) are known to play an important role in cancer pathogenesis, however, few information about their activity in human tumor tissues is available so far. We studied chymotrypsin-like activity of proteasomes in tissues of breast cancer (BC) and endometrial cancer (EC). The chymotrypsin-like total proteasome activity and the 20S and 26S proteasome activity in malignant tissues were shown to be significantly higher in malignant tumors than in normal tissues. No increase in proteasome activity was registered with larger tumor size in both BC and EC, whereas proteasome activity was changed with respect to the extent of tumor involvement. In breast cancer tissues, significant reductions in the total and the 26S proteaome activities were observed in tumors with regional lymph node metastases as compared to tumors without metastases. In endometrial cancer tissues, the total proteasome activity and the 20S and 26S proteasome activities were increased as the depth of myometrial invasion. The data obtained indicate that the proteasome acyivity is significantly changed in the process of cancerogenesis and further study is needed to develop new additional prognostic criteria and effective anti-tumor agents in molecular-directed therapy.

[Evaluation of risk of progression of endometrial hyperplasia in patients with metabolic syndrome].

A mathematical model based on principles of multifactor analysis was developed to predict clinical outcome of endometrial hyperplasia (EH) in patients with metabolic syndrome (80). Seventy-seven factors--anthropometric, clinical, anamnestic, hormono-metabolic, immunohistochemical, etc.--were included. Evaluation of the most informative indices integrated with the discriminative model showed that anthropometric (waist and hip circumference, sagittal diameter, etc.) and clinico-anamnestic (age, age of secondary sexual characters appearance, body weight at birth, suckling pattern, etc.) ones are of similar significance. A profile of hormono-metabolic parameters (cholesterol-low density lipoprotein, leptin, testosterone, progesterone and fasting glucose levels) helped identify a wide range of EH-related disorders in patients with metabolic syndrome. Consistently with the literature data, level of PTEN expression pointed to the presence of this tumor's suppressor in most EH cases which was matched by absence of its expression in endometrial carcinoma. Our model provided high sensitivity (89%) and specificity (82%) in predicting risk of progression in patients with endometrial hyperplasia and metabolic syndrome.

[Sex hormones in the systemic and local circulation in endometrial hyperplasia and neoplasia: correlation with the activity of estrogen-metabolizing enzymes].

Sex hormone profile was investigated in uterine systemic and local blood flow in endometrial hyperplasia and carcinoma. The results were compared with levels of aromatase and steroid sulfatase - estrogen metabolism enzymes. Local hyperestradiolemia and hyperestronemia were detected in cancer patients. Estradiol and estrone in endometrial hyperplasia were lower than in cancer which conforms to estrogen theory of hormonal carcinogenesis. There was no evidence for the role of hyperestrogenemia in endometrial hyperplasia development. Aromatase was a factor of intensified estrogen synthesis in altered and hyperplastic endometrium. There was an inverse correlation between intratumoral steroid sulfatase levels and those of estrone in uterine local blood flow.

[Prognostic significance of assay of sex hormones, their receptors and enzymes involved in estrogen synthesis and metabolism in endometrial carcinoma].

Prognostic significance of sex hormones, sex hormone-binding globulin, estrogen and progesterone receptors as well as the activity of enzymes involved in estrogen synthesis (aromatase, steroid sulfatase, estrogen hydroxylase, catechol-O-methyl transferase, glutathione-S-transferase) were investigated in 74 endometrial carcinoma patients. Stage and duration of uterine myoma history appeared to be significant factors of overall survival. Such factors as stage, depth of myometrial invasion, body mass, blood-serum estradiol level as well as aromatase and catechol-O-methyl transferase concentration in tumor tissue were significant for free-relapse free survival period.

Enzymes of estrogen metabolism in endometrial cancer.

Activities of estrogen metabolism enzymes (aromatase, 2- and 4-estrogen hydroxylases, catechol-O-methyltransferase, and glutathione transferase) were studied by modern biochemical methods in tumors of patients with endometrial cancer. Relationships between enzyme activities and body weight index, age of menarche, stage of the disease, tumor histotype, differentiation degree, and depth of invasion into the myometrium were detected. The detected relationships between enzyme activities and serum concentrations of estradiol and progesterone and level of estrogen receptors in tumor tissue attest to hormone dependence of aromatase, estrogen hydroxylases, and glutathione transferase.

Synthesis of highly tritium labeled steroids for evaluation of aromatase and steroidsulfatase activities in endometrial tumors.

Data on activities of estrogen-synthesis enzymes (aromatase and steroidsulfatase) in endometrial tumors were obtained by sensitive and highly precise radioisotope methods. The results clear out some pathogenetic aspects in the development and progress of malignant tumors in the endometrium.

[Clinical and morphologic aspects of cervical papillomavirus infection]. / Kliniko-morfologicheskie aspekty tservikal'noi papillomavirusnoi infektsii.

415 female residents of the City of Tomsk and Tomsk Region (patients with cervical carcinoma--22, dysplasia stage I-III--23 and healthy subjects--71) were screened for HPV16/18 infection, which was diagnosed in 18.3%. In the cervical carcinoma group, infection was detected in 27.4%, among patients with advanced cervical dysplasia--25.7%, and in those with background pathologies and healthy females--12.3 and 38%, respectively. Infection peaks were reported for the age brackets of 31-40 years (19.7%) and 51-60 years (19.6%). HPV-infection showed a wide range of colposcopic symptoms: areas of atypical blood vessels, leukoplakia, atypical epithelium and iodine-negative patches. Papillomavirus-related morphological changes in endometrial cells were typical of those associated with any viral infection and showed no specific features.