[Cloning of alternative isoforms of the catalytic subunit of the human thelomerase (htert)].

Most human tumors, including cervical cancer, are characterized by telomerase activation (cell proliferation activation enzyme). Such activation is implemented in the elongation of the terminal segments (telomeres) of the telomerase chromosome. The gene of the enzyme is RNA-encoded, the RNA in tumors being observed in a few isoforms. The hTERT RNA role in cell activation and control was simulated using cervical cancer, as well as its pretumoral states (CIN), as a model object. The goal of this work was to clone of the human hTERT isoforms (normal, α-, ß-, and α+ß-splice-variants). The genetic constructions containing normal hTERT sequence, α- and ß-deletion variants based on the lentivirus vector pR780 were obtained. The α- and ß-deletion variants were not obtained in this variant because of methodological problems. In further research, we plan to implement splice-variants of hTERT in eukaryotic human cells.

[The clinical morphological concept of origin and course of severe carcinoma in citu (CIN)].

The study was directed to determine objective causes of hypo-diagnostics of neoplastic process of cervix uteri with further development of clinical morphological concept close to understanding and comprehension by practical physicians to avoid diagnostic and tactical errors. The analysis was made concerning diagnostic errors and under-evaluation of degree of neoplasia in 880 patients with CIN of different degree and micro-invasive cancer of cervix uteri. The study also included 187 females with abnormal smears without CIN and cervix uteri cancer. All female patients underwent treatment in polyclinic of the N.N. Blokhin Russian oncological research center in 2006-2011. The correlation of data video-colposcopy pictures and morphology including misfits of results of repeated cytological and histological analysis of limited biopsies and materials of conizations. The study established that in 32% of patients ill-defined cells were detected only in material of endocervix. The rate of under-evaluation of severity of lesions in organic biopsy material increased with decreasing of visibility of transformation zone and increasing of age of patients. In case of fully visible transformation zone type I under-evaluation of degree of neoplasia consisted 12% and reached 20.7% in patients with transformation zone type II and 42.1% in patients with transformation zone type III. The visibility of transformation zone at age older than 30 years is often limited and this occurrence is established in all patents older than 35 years. The objective causes of hypo-diagnostics of neoplasia of cervix uteri in cytological smears and limited biopsy materials include transformation zone type II and especially type III and its age bias with transition zone into cervical channel with concealment there part of more "fresh" and severe lesions. The mentioned causes also embrace involvement of endocervical crypts into neoplastic process and their predominant lesion (in case of absence of ectopia) decreasing information value of smears from ectocervix and limited biopsies and polymorphism of affections as well.

[pAkt expression in cervical intraepithelial neoplasia (CIN) and in microinvasive cervical cancer].

The study objective was an immunohistochemical evaluation of pAkt expression in 81 CIN and microinvasive cervical cancer tissue samples and 10 samples of relatively "normal" cervical epithelium of HPV-infected women. PAkt expression showed significant up-regulation in CIN2, CIN3 and microinvasive cancer in compare to CIN1 and "normal" epithelium. The rate of pAkt- positive cells increased progressively by cervical neoplasia grade advancement reaching 7 +/- 5% in CIN2, 15 +/- 13% in CIN3 and 17 +/- 15% in microinvasive cancer. The rate of pAkt-positive cases in general was 1,7-fold higher in CIN3 (41%) than in CIN2 (24%). pAkt expression in conjunction with other markers may be used in immunohistochemical studies for individual CIN outcome prognosis and prospectively in immunocytochemical tests for CIN grade diagnostics improvement before using invasive methods. To elaborate multicomponent system of markers with their indexation there is a need for further investigations with greater number of cases.

[The role of cervical transformational zone as an object of human papilloma virus oncogenic effect in cervical intraepithelial neoplasms and invasive cancer development].

The correlation of morphological mistakes in neoplasia grade verification from visibility of transformation zone (TZ) and patient age was studied in 503 patients with CIN and microinvasive cervical cancer. The square of ectocervix lesion was defined by LeiseCap software in colposcopic working station Leisegang 3MV. The exclusive significance of TZ in HPV-associated cancerogenesis was confirmed clinically. We've established that the neoplasia stage increases with age while lesion extension and TZ visibility decrease dramatically leading to the subsequent decrease in colposcopy sensitivity and adequacy of guided biopsy. The critical age for underdiagnosis of latent lesions seems to be 35 years. The diagnostic and therapeutic value of conization and the large loop excision of the TZ (LLETZ) as the procedures with the optimal TZ excision in patients with visibly unchanged ectocervix are confirmed in cases when CIN 2-3 and microinvasive cancer are suspected.

[Value of HPV (hybrid capture 2) testing for assessing the effectiveness of conservative surgical treatment in CIN2-3/carcinoma in situ and microinvasive cervical cancer].

The prospective study in 525 patients with CIN2-3/CIS and microinvasive cervical cancer showed the association of epithelial lesions with HPV HR in almost all cases. This allowed using HPV-testing for conservative surgical treatment (ablation, LEEP/ LLETZ, conization) efficacy assessment and patient follow-up. The value of HPV-testing for residual disease and recurrence detection was higher that of cytology and colposcopy. HPV-testing showed 100% sensitivity and negative predictive value. Hybrid Capture 2 test was more useful for residual disease and recurrence detection than PCR, showed higher specificity (96% vs. 79,2%) and positive predictive value (44.8% vs. 13.4%). After conservative surgical treatment of CIN2-3/carcinoma in situ and microinvasive cervical cancer Hybride Capture 2 is to become negative thus confirming the adequacy of neoplastic tissue excision.

[Ki-67 expression, thymidine phosphorylase and PTEN in intraepithelial cervical carcinoma].

Expression of Ki-67, thymidine phosphorylase (TP) and PTEN were assessed in various grades of cervical intraepithelial neoplasia (CIN) in order to evaluate their potentials of predicting the gravity of possible damage to the epithelium as well as pro- or regression of CIN. Ki-67 and TP levels were shown to correlate directly with CIN grade. It was suggested that a small number of cases of Ki-67 and TP expression absence (15%), exclusively in CIN3 samples, be due to imminent progression to invasive cancer. Both separately and in combination, Ki-67 and TP expression indices should be regarded as having a potential as markers for cervical carcinoma diagnosis, grade and clinical course.

[Are human papillomaviruses responsible for the occurrence of bladder cancer].

A female patient with recurrent bladder cancer underwent complex examination. The primary tumor removed in 2004 showed human papillomavirus (HPV) 16 DNA, mRNA corresponding to HPV16 oncogene E7, as well as HPV16 protein E7. The patient is a smoker who has been working at a chemical factory for over 20 years. During tumor recurrence in 2009, there was no DNA of high-risk HPV types in the cancer cells. HPV16 E7protein and cellular p 16(INK4alpha), an indicator of HPV-induced carcinogenesis, were not found. Colposcopy revealed no precancerous changes in the epithelium of the cervix uteri. The cervical epitheliocytes contained no high-risk HPV DNA, E7 and p16(INK4alpha) proteins. It seems expedient to continue in vitro studies of the possible role of HPV in urothelial carcinogenesis on an experimental model.

[Clinico-morphological characteristics of light-cell adenocarcinoma of the ovaries]. / Kliniko-morfologicheskie osobennosti svetlokletochnykh adenokartsinom iaichnikov.

Clinical and morphological data on 37 observations of a rare neoplasm, clear-cell adenocarcinoma of the ovaries (CAO), are presented. Studies of the microstructure permitted the authors to distinguish 4 histological variants: tubular, papillary, solid, and mixed. Comparison of the data on the three-year survival rate with different histological variants of the tumor in all stages and within stage I revealed a higher survival rate in patients with the tubular variant of CAO which is considered to be more favourable prognostically. The data of 3-year survival rate of all the patients (32.8 +/- 7.9%), and of those in stage I, with the tubular variant (88.9 +/- 10.5%), papillary and mixed variants taken together (23 +/- 11.5%), are presented. In the latter the survival rate is much lower than in the tubular variant of the tumor. The variant of the CAO microstructure should be included into the diagnosis.

[Clear cell adenocarcinoma of the corpus uteri]. / Svetlokletochnaia adenokartsinoma tela matki.

Specimens obtained from 15 patients suffering from a rare form of carcinoma of corpus uteri--a clear cell adenocarcinoma--were studied. The importance of identification of this form of cancer as a specific nosologic unit was shown, since the 5-year survival rate in patients with this lesion (57.2 +/- 16.4%) was lower than that in patients with endometrial adenocarcinoma, involvement in stage I tumor patients and scheme of treatment being identical. The clear cell adenocarcinoma was shown to occur in more elderly patients as compared with adenocarcinoma of the endometrium and its symptoms are not pathognomonic. The occurrence, in some cases, of clear cell adenocarcinoma in the endometrium and co-occurrence of the elements of endometrial and clear cell adenocarcinomas in the same tumor point to the origin of clear cell adenocarcinoma from the elements of Müller's system.

Cervical intraepithelial neoplasia: Telomerase activity and splice pattern of hTERT mRNA.

Cervical cancers are characterized by the persistence of human papilloma virus (HPV) genome that is found in tissue samples starting from the early stages of tumor progression. Just like in other tumors, the activation of telomerase was observed in cervical carcinomas, but information about its expression was controversial. The aim of this study is to find possible correlations between the presence of HPV sequences, activity of telomerase and expression of different spliced forms of hTERT RNA in cervical intraepithelial neoplasias (CIN). The results show that HPV DNA is present in 60% of normal tissue adjacent to CIN lesions and up to 84% in CIN samples. Telomerase activity was found in 28% of adjacent normal tissue and in 68% of CIN II-III. hTERT RNA that encodes an active enzyme was present almost in all CIN samples. Variations in levels of telomerase activity are possibly not regulated by the splicing forms of hTERT mRNA with deletions.