Induction of Integrin Signaling by Steroid Sulfatase in Human Cervical Cancer Cells

Steroid sulfatase (STS) is an enzyme responsible for the hydrolysis of aryl and alkyl sulfates. STS plays a pivotal role in the regulation of estrogens and androgens that promote the growth of hormone-dependent tumors, such as those of breast or prostate cancer. However, the molecular function of STS in tumor growth is still not clear. To elucidate the role of STS in cancer cell proliferation, we investigated whether STS is able to regulate the integrin signaling pathway. We found that overexpression of STS in HeLa cells increases the protein and mRNA levels of integrin β1 and fibronectin, a ligand of integrin α5β1. Dehydroepiandrosterone (DHEA), one of the main metabolites of STS, also increases mRNA and protein expression of integrin β1 and fibronectin. Further, STS expression and DHEA treatment enhanced phosphorylation of focal adhesion kinase (FAK) at the Tyr 925 residue. Moreover, increased phosphorylation of ERK at Thr 202 and Tyr 204 residues by STS indicates that STS activates the MAPK/ERK pathway. In conclusion, these results suggest that STS expression and DHEA treatment may enhance MAPK/ERK signaling through up-regulation of integrin β1 and activation of FAK.

Two Cases of Pre-descemet Corneal Dystrophy Associated with X-linked Ichthyosis: A Case Report by Genetic Analysis

PURPOSE: To report the first case of steroid sulfatase (STS) gene deletion, confirmed by multiplex ligation-dependent probe amplification (MLPA) analysis in identical twins with pre-Descemet corneal dystrophy associated with X-linked ichthyosis. CASE SUMMARY: 19-year old identical twin brothers with itching senses and hereditary thick scaly skin of the extremity and trunk visited our dermatologic clinic. Upon visiting, an ophthalmologic consultation with anterior segment examination showed diffuse punctate corneal opacities in the pre-Descemet layer. On MLPA analysis of the identical twin brothers, a definitive diagnosis of X-linked ichthyosis was made by identifying STS gene deletion. CONCLUSIONS: Identification of the deletion and mutation of the involved gene using gene analysis can provide insight to diagnosis and clinical characteristics of X-linked ichthyosis.

Identification of gene mutation and prenatal diagnosis in a family with X-linked ichthyosis / 中国当代儿科杂志

X-linked ichthyosis (XLI) is a metabolic disease with steroid sulfatase deficiency and often occurs at birth or shortly after birth. The encoding gene of steroid sulfatase, STS, is located on the short arm of the X chromosome, and STS deletion or mutation can lead to the development of this disease. This study collected the data on the clinical phenotype from a family, and the proband, a boy aged 11 years with full-term vaginal delivery, had dry and rough skin and black-brown scaly patches, mainly in the abdomen and extensor aspect of extremities. Peripheral blood samples were collected from each family member and DNA was extracted. Multiplex ligation-dependent probe amplification (MLPA) was used to measure the copy number of STS on the X chromosome. Whole-genome microarray was used to determine the size of the segment with microdeletion in the X chromosome. MLPA was then used for prenatal diagnosis for the mother of the proband. The results revealed that the proband and another two male patients had hemizygotes in STS deletion. Gene microarray identified a rare deletion with a size of 1.6 Mb at Xp22.31 (chrX: 6,516,735-8,131,442). Two female family members were found to be carriers. Prenatal diagnosis showed that the fetus carried by the proband's mother was a carrier of this microdeletion. This study showed STS gene deletion in this family of XLI, which causes the unique skin lesions of XLI. MLPA is a convenient and reliable technique for the molecular and prenatal diagnosis of XLI.

The Role of Steroid Sulfatase as a Prognostic Factor in Patients with Endometrial Cancer

PURPOSE: The aim of the study was to determine steroid sulfatase (STS) expression in endometrial cancer patients and its correlation with disease prognosis. MATERIALS AND METHODS: We conducted a retrospective study in 59 patients who underwent surgery with histologically confirmed endometrial cancer from January 2000 to December 2011 at Hanyang University Hospital. Immuno-histochemical staining of STS was performed using rabbit polyclonal anti-STS antibody. RESULTS: Sixteen of the 59 patients (27.1%) were positive for STS expression. Disease free survival (DFS) was 129.83±8.67 [95% confidence interval (CI): 112.84-146.82] months in the STS positive group (group A) and 111.06±7.17 (95% CI: 97.01-125.10) months in the STS negative group (group B) (p=0.92). Overall survival (OS) was 129.01±9.38 (95% CI: 110.63-147.38) months and 111.16±7.10 (95% CI: 97.24-125.07) months for the groups A and B, respectively (p=0.45). Univariate analysis revealed that FIGO stage and adjuvant therapy are significantly associated with DFS and OS. However, in multivariate analysis, FIGO stage and adjuvant therapy did not show any statistical significance with DFS and OS. STS was also not significantly associated with DFS and OS in univariate and multivariate analysis. CONCLUSION: STS expression was not significantly associated with DFS and OS, despite positive STS expression in 27% of endometrial cancer patients. Therefore, the role of STS as a prognostic factor in patients with endometrial cancer remains unclear and requires further research.

Induction of steroid sulfatase expression by tumor necrosis factor-alpha through phosphatidylinositol 3-kinase/Akt signaling pathway in PC-3 human prostate cancer cells

Steroid sulfatase (STS) is responsible for the hydrolysis of aryl and alkyl steroid sulfates and has a pivotal role in regulating the formation of biologically active estrogens. STS may be considered a new promising drug target for treating estrogen-mediated carcinogenesis. However, the molecular mechanism of STS expression is not well-known. To investigate whether tumor necrosis factor (TNF)-alpha is able to regulate gene transcription of STS, we studied the effect of TNF-alpha on STS expression in PC-3 human prostate cancer cells. RT-PCR and Western blot analysis showed that TNF-alpha significantly induced the expression of STS mRNA and protein in a concentration- and time-dependent manner. Treatment with TNF-alpha resulted in a strong increase in the phosphorylation of Akt on Ser-473 and when cells were treated with phosphatidylinositol (PI) 3-kinase inhibitors such as LY294002 or wortmannin, or Akt inhibitor (Akt inhibitor IV), induction of STS mRNA expression by TNF-alpha was significantly prevented. Moreover, activation of Akt1 by expressing the constitutively active form of Akt1 increased STS expression whereas dominant-negative Akt suppressed TNF-alpha-mediated STS induction. We also found that TNF-alpha is able to increase STS mRNA expression in other human cancer cells such as LNCaP, MDA-MB-231, and MCF-7 as well as PC-3 cells. Taken together, our results strongly suggest that PI 3-kinase/Akt activation mediates induction of human STS gene expression by TNF-alpha in human cancer cells.

Ictiosis ligada al cromosoma X: comunicación de tres casos y revisión de la literatura / X-linked ictiosis: report of 3 cases and review of the literature

La ictiosis ligada al cromosoma X es una genodermatosis poco frecuente que afecta a varones a partir de las 2 semanas de vida y se caracteriza por xerodermia, hiperqueratosis, descamación y, en algunos casos, manifestaciones extracutáneas como criptorquidia y opacidad de la córnea. La enfermedad es de evolución crónica y experimenta mejoría parcial en época estival. La deleción total o parcial de la enzima sulfatasa esteroidea es la causa de las manifestaciones clínicas. Presentamos tres pacientes con ictiosis ligada al cromosoma X, a quienes se les realizaron interconsultas con los servicio de Oftalmología y Pediatría. Se encuentran actualmente en tratamiento con emolientes...

Nephrotic syndrome with X-linked ichthyosis, Kallmann Syndrome and unilateral renal agenesis.

We describe a 10-year-old boy with X-linked ichthyosis, Kallmann Syndrome and unilateral renal agenesis who presented with nephrotic syndrome. DNA analysis revealed deletion of the Steroid Sulfatase (STS) gene. STS deficiency in X-linked ichthyosis leads to cholesterol sulfate accumulation, which induces transglutaminase-1 dysfunction. Since the slit diaphragm of the glomerular epithelial cell is a modified adherens junction, the accumulation of cholesterol sulfate could interfere with the normal slit diaphragm function of the glomerular visceral epithelial cell, resulting in nephrotic range proteinuria. The child went into remission on oral prednisolone.

STS gene in a pedigree with X-linked ichthyosis / 华中科技大学学报（医学）（英德文版）

To investigate the gene mutation in a pedigree with X-linked ichthyosis (XLI) and to explore the relationship between the mutation and its clinical manifestations, genomic DNA of affected members, the normal member of the pedigree and 50 unrelated normal members was extracted with a whole blood genomic DNA extraction kit and the DNA was used as a template for the polymerase chain reaction (PCR)-mediated amplification of exon 1 and exon 10 of the STS gene. hHb6 (human hair basic keratin) gene was used as the internal control. Our results showed that the STS gene was deleted in affected members in the pedigree with X-linked ichthyosis. The normal member of the pedigree and 50 unrelated normal members had no such deletion. The proband and his mother had products in the internal control after PCR amplification. The blank control had no product. It is concluded that deletion of the STS gene existed in this pedigree with X-linked ichthyosis, and it is responsible for the unique skin lesions of X-linked ichthyosis.

Cell Cycle and Gene Expression of Adriamycin-Induced Apoptosis in Human Cervical Carcinoma HeLa cells / 대한산부인과학회잡지

OBJECTIVE: This work was demonstrated the induction of apoptosis in response to adriamycin, we checked the cell cycle of adriamycin-induced apoptosis and to investigate whether differential expression is associated with adriamycin-induced genes in human cervical carcinoma HeLa cells. METHODS: Apoptosis was measured by flow cytometry for cell cycle analysis in Hela cells. Differential expression is associated with adriamycin-induced genes in HeLa cells, it was performed to purifiy the RNA, cDNA probe and hybridization. The various different overexpressed genes were determined by gene array analysis (GDA). Analysis were referenced Incyte Genomics Co. (http://www.ncbi.nlm.nih.gov/). RESULTS: We found that adriamycin was induced apoptosis in a dose- and time-dependent manner, as demonstrated by sub-G0/G1 peaks in DNA content histogram of cell cycle. The cells of G2/M phase by treatment of 0.1 microgram/mL adriamycin had been arrested. G2/M peaks in DNA content was decreased in a dose and time-dependent manner. It had been observed 6 group, 16 genes. The group I contained thioredoxin and cytochrome c oxidase subunit IV gene, group II were p53 and excision repair protein (ERCC-1) gene. Group III was metabolic regulated gene, glucosidase, AMP deaminase isoform L (AMPD2), glutamine synthetase, cholesterol 25-hydroxylase, and steryl-sulfatase precursor. Group IV was cell skeleton constructed gene, heparan sulfate proteoglycan (HSPG2), and microfibrillar-associated protein (MFAP2), group V was oncogene group, v-yes-1 Yamaguchi sarcoma viral oncogene homolog-1 (YES1) and tyrosine kinase ELK1. The other group 6 contained NOD1 protein gene interleukine-1 receptor accessory protein (IL1RAP), pregnancy-specific glycoprotein-11 (PSG11), and pregnancy-specific protein-1a (PSG-1a). CONCLUSION: The present findings indicating that adriamycin was revealed apoptosis in Hela cell. Differential gene expression is related in various metabolism by adriamycin.

Cell Cycle and Gene Expression of Adriamycin-Induced Apoptosis in Human Cervical Carcinoma HeLa cells / 대한산부인과학회잡지

OBJECTIVE: This work was demonstrated the induction of apoptosis in response to adriamycin, we checked the cell cycle of adriamycin-induced apoptosis and to investigate whether differential expression is associated with adriamycin-induced genes in human cervical carcinoma HeLa cells. METHODS: Apoptosis was measured by flow cytometry for cell cycle analysis in Hela cells. Differential expression is associated with adriamycin-induced genes in HeLa cells, it was performed to purifiy the RNA, cDNA probe and hybridization. The various different overexpressed genes were determined by gene array analysis (GDA). Analysis were referenced Incyte Genomics Co. (http://www.ncbi.nlm.nih.gov/). RESULTS: We found that adriamycin was induced apoptosis in a dose- and time-dependent manner, as demonstrated by sub-G0/G1 peaks in DNA content histogram of cell cycle. The cells of G2/M phase by treatment of 0.1 microgram/mL adriamycin had been arrested. G2/M peaks in DNA content was decreased in a dose and time-dependent manner. It had been observed 6 group, 16 genes. The group I contained thioredoxin and cytochrome c oxidase subunit IV gene, group II were p53 and excision repair protein (ERCC-1) gene. Group III was metabolic regulated gene, glucosidase, AMP deaminase isoform L (AMPD2), glutamine synthetase, cholesterol 25-hydroxylase, and steryl-sulfatase precursor. Group IV was cell skeleton constructed gene, heparan sulfate proteoglycan (HSPG2), and microfibrillar-associated protein (MFAP2), group V was oncogene group, v-yes-1 Yamaguchi sarcoma viral oncogene homolog-1 (YES1) and tyrosine kinase ELK1. The other group 6 contained NOD1 protein gene interleukine-1 receptor accessory protein (IL1RAP), pregnancy-specific glycoprotein-11 (PSG11), and pregnancy-specific protein-1a (PSG-1a). CONCLUSION: The present findings indicating that adriamycin was revealed apoptosis in Hela cell. Differential gene expression is related in various metabolism by adriamycin.

Diagnosis of x-linked ichthyosis and detection of its carriers with southern blot hybidization / 대한피부과학회지

BACKGROUND: The skin changes of X-linked recessive ichthyosis are cnused by the deficiency of the enzyme steroid sulfatase, which usually results from deletions of this gene in Caucasian populations. OBJECTIVE AND MEHTODS: To disgnose X-linked recessive ichthyosis and detect its carrier, we have investigated distinctive gene deletion and measured gene dosage of steroid sulfatase gene by southern blot hybridization in Korean patients with X-linked recessive ichthyosis. RESULTS: Patients from 8 of 9 unrelated families exhibited deletions, if the steroid sulfatase gene. Of 6 families showing a family history compatible with X-linked recessive inheritance, One family exhibited a normal pattern of hybridization. All but one family showed deletion of steroid sulfatase gene. All three patients lacking a fami1y history of the disease exhibited gene deletions. The ratio of the steroid sulfatsse specific band density to the Factor VIII specific band density was measured in 8 obligate carriers using a laser densitometer. The average ratio exhibited by the car riers was less than half that of normal women. Conclusian: These results suggest that the X-linked recessive ichth osis patient and its carrier can also be diagnosed and detected by Southern blot hybridization of steroid sulfatase gene in Korea.