Role of estrogen receptors in health and disease.

Estrogen receptors (ERs) regulate multiple complex physiological processes in humans. Abnormal ER signaling may result in various disorders, including reproductive system-related disorders (endometriosis, and breast, ovarian, and prostate cancer), bone-related abnormalities, lung cancer, cardiovascular disease, gastrointestinal disease, urogenital tract disease, neurodegenerative disorders, and cutaneous melanoma. ER alpha (ERα), ER beta (ERß), and novel G-protein-coupled estrogen receptor 1 (GPER1) have been identified as the most prominent ERs. This review provides an overview of ERα, ERß, and GPER1, as well as their functions in health and disease. Furthermore, the potential clinical applications and challenges are discussed.

[Expression of fusion gene PAX3/PAX7-FKHR and chromosomal aberration in rhabdomyosarcoma].

OBJECTIVE: To detect the PAX3/PAX7-FKHR fusion transcripts to identify genetic alteration in embryonal rhabdomyosarcoma (ERMS) and alveolar rhabdomyosarcoma (ARMS) tissues. METHODS: One-step reverse transcription- polymerase chain reaction (RT-PCR) were used to detect the expression of the PAX3/PAX7-FKHR fusion transcrips in 16 cases of rhabdomyosarcoma (7 cases of ARMS, 9 cases of ERMS) and 16 specimens were compared to the surrounding normal tissue. Comparative genomic hybridization (CGH) was employed to detect the genomic imbalance (DNA loss or amplification) in 16 RMS cases. RESULTS: PAX3-FKHR fusion transcripts were positive in 3/7 and PAX 7-FKHR fusion transcripts were positive in 2/7 of ARMS patients, respectively, and were all negative in ERMS and Control tumors. There were different chromosome variations for each RMS, chromosome amplification was frequently seen in 1p36 (69%), 5q32 (56%), 8q21 (63%), 13q14 (69%), 19q (63%), 20q (56%). Chromosome loss was frequently seen in 3p21-pter (56%), 9p23-pter (50%), 10q (69%), 16/16q24 (56%). CONCLUSION: One-step RT-PCR assay for detection specific fusion gene provides a useful tool for confirmation of the diagnosis of RMS in diagnostically difficult cases and in retrospective studies. Chimeric gene transcript resulting from specific chromosomal translocations is a reliable index for the molecular diagnosis of RMS.

[Genomic imbalance and chromosome disorders in hepatoblastoma].

OBJECTIVE: To establish stable techniques of comparative genomic hybridization (CGH) and apply them to elucidate the genetic characteristics of hepatoblastoma (HB), and to explore the characteristics and clinical significance of loss of heterozygosity (LOH) at 1p36 in HB. METHODS: CGH was employed to detect the genomic imbalance (DNA loss or amplification) in 20 cases of HB, and PCR-simple repeated sequence polymorphism was employed in 30 cases of HB to detect the loss of heterozygosity for 6 satellites at chromosome 1p36. RESULTS: There were different chromosome variations for each HB. chromosome amplification was frequently seen in 1q, 2q,2p, 8q, 8p, 12q and 22q. Chromosome loss was often seen in 1p, 4q, 4p, 16q, 17p and 18q. The frequency of LOH at 6 loci on chromosome 1 was 63.3% totally (19/30), with the highest D1S199 (66.7%) and D1S450 next to it (46.7%). CONCLUSION: There were chromosome zones with DNA amplification or loss in hepatoblastoma. There are extensive LOH at 1p36 in hepatoblastoma. The corresponding amplification of oncogene and loss of antioncogene may take part in the development of hepatoblastoma.

Specific siRNA inhibits XIAP expression in human endometrial carcinoma cell apoptosis.

The aim of the study was to investigate the inhibitory effects of RNA interference on XIAP gene expression of human endometrial carcinoma RL95-2 cell and the cell apoptosis. Specific small interference RNA (siRNA) of XIAP was designed and composed. Transfection of siRNA was conducted in endometrial carcinoma cell line RL95-2. The XIAP gene mRNA was assessed by real-time PCR and the change of XIAP protein was assessed with Western Blotting. The cell proliferation and apoptosis was assessed by MTT and flow cytometry methods. After transfection of siRNA specifically targeting XIAP, the relative fold of mRNA transfection in the specific transfection group was (0.04 ± 0.06) and the relative protein expression was (0.590 ± 0.178), which was significantly decreased when compared with the control group (P < 0.05); the cell growth inhibition rate in the transfection group was (47.86 ± 4.46)%, which was significantly increased when compared to the control group (P < 0.05). In vitro experiment showed that synthetic siRNA could effectively inhibit the transfection and expression of XIAP gene of human endometrial carcinoma RL95-2 cell at the mRNA level and protein level, thus significantly promote the apoptosis of endometrial carcinoma cell. The mechanisms involved in the apoptosis still require further investigation.

miR-200c modulates ovarian cancer cell metastasis potential by targeting zinc finger E-box-binding homeobox 2 (ZEB2) expression.

This study was to investigate the effect of miR-200c on regulation of ovarian cancer cell metastasis potential and explore the underlying molecular events. qRT-PCR was used to analyze the level of miR-200c expression in 48 ovarian cancer and 30 normal ovarian tissue samples. pre-miR-200c was used to manipulate miR-200c expression in ovarian cancer cells for detection of changed phenotypes of tumor cells. Bioinformatics analysis was then used to predict target genes of miR-200c and GO and pathway analyses drew the miR-200c-related gene network. Luciferase reporter assay confirmed the target of miR-200c as ZEB2. Western blot was used to detect gene expressions in ovarian cancer cells. Level of miR-200c expression was much higher in ovarian cancer than in normal ovarian tissues, and miR-200c expression was inversely associated with advanced clinical stage and lymph node metastasis of ovarian cancer (p < 0.01). The database search predicted 186 miR-200c-targeting genes, and GO analysis showed that functions of these target genes were enriched in the protein binding and other biological processes. Furthermore, miR-200c expression inhibited ovarian cancer cell ES-2 migration and invasion capacity by suppression of ZEB2 expression (p < 0.01). Overexpression of miR-200c regulated E-cadherin and vimentin expression in ovarian cancer cells. This study demonstrated high miR-200c expression in ovarian cancer tissues and ZEB2 as a targeting gene of miR-200c, which mediated the effects of miR-200c on regulation of ovarian cancer cell migration and invasion capacity and epithelial-to-mesenchymal transition. Thus, targeting of miR-200c or ZEB2 may serve as a potential therapeutic strategy for control of ovarian cancer.

[The infection of dendritic cells by recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and its biological characteristics observations].

OBJECTIVE: To test the infeciton efficiency of recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and to abserve its biological characteristics. METHODS: Peripheral blood mononuclear cells (PBMC) were separated from healthy blood donor and they were infected by Ad-HSP70-HBsAg on the first day of isolation. DCs were induced in medium with cytokines IL-4, GM-CSF and TNF-alpha in vitro. The biological characteristics of DC induced were analyzed by inverted fluorecent microscope, RT-PCR, flow cytometer (FACS), and mixed lymphocyte reaction (MLR). RESULTS: The traced gene-GFP were abserved in DCs by inverted fluorecent microscope and HSP70-HBsAg gene mRNA expression was detected by RT-PCR after the Ad-HSP70-HBsAg infection. FACS analysis shown that the expression of CD1a, CD80, CD86 and HLA-DR on surfece of two groups of DCs were similar. MLR showed that there are not a statitic difference of stimulated index (SI) between two groups. CONCLUSION: Results indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics.

[alpha1,2-fucosyltransferase gene transfection influences on biological behavior of ovarian carcinoma-derived RMG-I cells].

To explore the effect of alpha1,2-fucosyltransferase (alpha1,2-FT) gene on the biological behavior of ovarian carcinoma cells, the expression vector pcDNA3.1 -HFUT-H which containing human alpha1,2-FT gene coding region was constructed and transfected into ovarian carcinoma cell line RMG-I. alpha1,2-FT gene expression, Lewis y antigen expression, alpha1,2-FT activity, rates of cell growth and tumorogenesis in nude mice were compared before and after RMG-I being transfected. The transfection of alpha1,2-FT gene significantly increased the expression of alpha1,2-FT gene and Lewis y antigen in RMG-I cells. After the transfection, RMG-I cells showed remarkable improvement of alpha1,2-FT activity, cell proliferation, the ability of colony formation in soft agar and tumorogenesis in nude mice. The growth rates of alpha1,2-FT gene transfected RMG-I cells were obviously increased, presenting that the proportion of the cells in G2-M and S phases was obviously increased from 2.05% to 7.32% and from 23.95% to 33.27% respectively, and that of the cells in G1 phase was obviously decreased from 74.14% to 59.46%, as compared with the control group. These findings indicate that alpha1,2-FT and Lewis y antigen have the ability to enhance the proliferation and elevate the survival rates of RMG-I cells, which can promote the genesis and development of ovarian carcinoma.