[Clinical Findings in Two patients with DSD 46XY caused by new variant of the Desert Hedgehog Gene and review of the literature of the role of DHH signaling pathway in sex development].

Mutations in the gene DHH are an extremely rare cause of disorders of sex development 46,XY (DSD,46XY). The article describes the clinical cases of two unrelated patients with gonadal dysgenesis 46,XY with female phenotype. By using a next generation sequencing method, in both cases the same biallelic variant substitution c. 419T>G in the DHH gene was revealed. Taking into account the data on the role of DHH in the formation of the nervous system, the diagnosis of minifascicular polyneuropathy at the preclinical stage was confirmed in both cases. These cases demonstrate the value of using NGS, which allows simultaneous analysis of a wide range of candidate genes in DSD and the diagnosis of comorbidities before the development of the clinical picture. These are the first descriptions of patients with mutations in the DHH gene in the Russian population.

[Bcl-2 family proteins and mechanisms of the acceleration and inhibition of tumor progression in vivo].

Earlier the authors demonstrated that the process of tumor progression in vivo may be inhibited or accelerated depending on the conditions of tumor growth (accelerated by tumor cell dissemination or delayed in locally growing tumors). It was also shown that tumor progression is inhibited in case of bcl-2 gene transduction in tumor cells. In this study, the research into mechanisms of the acceleration or inhibition of tumor progression and the role that Bcl-2 family proteins may play in these phenomena was continued. The results of the study demonstrated the following 1) immediate in vivo activation of endogenous proapoptotic Bax protein in disseminated tumor cells, not protected by Bcl-2 against apoptosis, and its correlation with accelerated tumor progression; 2) complete suppression of in vivo Bax activation in tumor cells protected by Bcl-expression, and inhibited tumor progression; 3) alternative character of Bcl-2 and Bax expression under the conditions of accelerated and inhibited tumor progression. Thus, the data presented support the hypothesis that the rates of tumor progression in vivo are regulated depending on the initial anti- and proapoptotic programs of tumor cells.

Cell transforming genes and tumor progression: in vivo unified secondary phenotypic cell changes.

We have earlier shown that Syrian hamster cells spontaneously transformed in vitro during in vivo progression, acquire in 1 step, along with highly increased tumorigenicity, 2 new properties characterizing the [H2O2CA + tPGE(S)] phenotype, i.e., a high H2O2 catabolizing (antioxidant) activity and the ability to release PGE2 upon contact with NK cells. In contrast, RSV-SR-(v-src)-transformed cells acquire the [H2O2CA + PGE(S)] phenotype and high tumorigenicity during in vitro transformation, i.e., without preliminary in vivo selection. In the present study, the possible influence of different transforming genes on the rates of subsequent in vivo tumor progression was studied using cells in vitro transformed by SV40, BAV-3, or transduced by activated genes Ha-ras, p53, myc and bcl-2. The expression of the [H2O2CA + PGE(S)] phenotype, the extent of tumorigenic and spontaneous metastasizing activities were examined before and during in vivo cells selection in s.c. growing tumors. Our results demonstrate that: (1) after in vitro transformation all cell lines (except v-src) were negative for the expression of [H2O2CA + PGE(S)] phenotype and remained equally low-tumorigenic; (2) independently of the types of genes initially transforming the cells, in vivo tumor progression was consistently leading to the replacement of parental cells by cells expressing the [H2O2CA + PGE(S)] phenotype, to 30-200 times increased tumorigenicity and less frequently to metastasizing; (3) the time necessary for selection of cells expressing this phenotype was the same (about 180 days in vivo) for all transformants, except bcl-2; the latter reaching similar values after a significant delay. Thus, common secondary src-like phenotypic cell changes, regardless of initially cell transforming genes are necessarily selected during tumor progression in vivo.

Allogenic bone-marrow transfusion suppresses development of lung metastases in osteogenic sarcoma patients after radical surgery.

On the basis of experimental data obtained in Syrian hamsters, demonstrating the highly efficient suppression of experimental and spontaneous metastases of highly-metastatic sarcoma cells by the use of allogeneic normal bone-marrow cells (BMC), a clinical protocol for the suppression of lung metastases of osteogenic sarcoma was started in 1984 in the Cancer Research Center, Moscow. From this time onwards, 24 osteogenic sarcoma patients, at stages 2A and 2B were treated with a combination of radical surgery and a single transfusion of normal (non-activated) allogeneic BMC (blood-group and Rhesus compatible). The first results of this ongoing study are now presented. Metastases appeared in 11 out of the 24 patients, generally very early during the first 3-9 months after treatment and in no case after 2 years. More than 50% of the BMC-treated patients were free of lung metastases after 2 or more years of observation; 8 out of 15 are still metastasis-free after 3-4 or more years of observation following treatment. The differences in the frequency of metastasis and duration of survival without metastasis of treated patients compared with a group of 41 osteogenic sarcoma patients at stages 2A and B, treated with radical surgery only (controls) reached significant levels 12 months after treatment and thereafter. Rapid recovery of NK cytotoxic activity has been observed in nearly all successfully BMC-treated patients.

[The role of prostaglandins type E in tumor cells during their contact with NK-cells in vitro]. / Rol' prostaglandinov tipa E v opukholevykh kletkakh pri ikh kontakte s NK-kletkami in vitro.

Active PGE--secretion by malignant tumour cells of Syrian hamsters was demonstrated 30 min after their contact with NK-cells in vitro. The duration of PGE--secretion depended upon the ratio of tumour cells and NK--cells, engaged in contact. Increase of the number of NK--cells (bound to tumour cells) up to 10:1-20:1 led to rapid release of PGE from majority of tumour cells; in this case PGE release was continued not longer than 1.5-2.0 hours. The active release of PGE can be stopped after the contact of tumour and NK--cells by indomethacin at any moment of its secretion.

Clustering of discrete cell properties essential for tumorigenicity and metastasis. III. Dissociation of the properties in N-ras-transfected RSV-SR-transformed cells.

We have previously shown that RSV-SR-transformed hamster cells acquire high resistance to H2O2, i.e. the cytotoxic product of activated macrophages (H2O2R) and that they begin to secrete PGE (PGES), thus inactivating the CTA of NK cells. Among normal cells, the same phenotype is expressed in activated macrophages. In all our RSV-transformed cells these 2 properties were jointly expressed and correlated with high tumorigenicity and experimental metastasizing of these cells. We now show that transfection of 3 RSV-SR-transformed cell strains with activated N-ras leads either to complete inhibition of the H2O2R + PGES phenotype in all clones of one strain, or to inhibition of PGES only in the majority of clones of 2 other strains. Unexpectedly, the complete or partial inhibition of this phenotype did not alter the high tumorigenicity of 2 strains of these cells, but lower tumorigenicity was evident in almost all clones of the third strain (as well as in some gene-neo-transfected clones of these strains). The loss of PGES made these cells susceptible to the CTA of NK cells, while the loss of H2O2R did not alter their resistance to the CTA of macrophages. Expression of the H2O2R + PGES phenotype was retained in all cloned variants of control, gene-neo-transfected cells. The possible relation of the N-ras gene to regulation of src gene activities in RSV-SR-transformed cells is discussed.