Hye-Sung Moon

Matrix metalloproteinase-2, membranous type 1 matrix metalloproteinase, and tissue inhibitor of metalloproteinase-2 expression in ectopic and eutopic endometrium

OBJECTIVE To investigate expression of matrix metalloproteinase-2 (MMP-2), membranous type 1 matrix metalloproteinase (MT1-MMP), and tissue inhibitor of metalloproteinase-2 (TIMP-2) in ectopic and eutopic endometrium from women with and without endometriosis throughout the menstrual cycle. DESIGN Molecular studies in human tissue. SETTING Reproductive immunology laboratory of a university medical center. PATIENT(S) Fifty-three premenopausal woman (23 with endometriosis and 30 without endometriosis) undergoing laparoscopic surgery. Endometrium and ectopic endometriosis tissue were obtained at the time of surgery. MAIN OUTCOME MEASURE(S) Messenger RNA and protein expression from eutopic and ectopic endometrium was analyzed by using quantitative competitive polymerase chain reaction, zymography, and Western blot assay. RESULT(S) Uterine endometrium from women with endometriosis expressed higher levels of MMP-2 and MT1-MMP and lower levels of TIMP-2 than did endometrium from normal women. CONCLUSION(S) Eutopic endometrium from patients with endometriosis may be more invasive and prone to peritoneal implantation because of greater expression of MMP-2 and MT1-MMP and lower expression of TIMP-2 messenger RNA, compared with endometrium from women without endometriosis. Thus, increased proteolytic activity may help to explain the invasive factors that result in endometriosis.

Effects of the GSTM1 and GSTT1 polymorphisms on the relationship between maternal exposure to environmental tobacco smoke and neonatal birth weight.

The purpose of the investigation was to determine whether genetic polymorphisms in enzymes that metabolize exogenous chemicals modulate the effects of environmental tobacco smoke (ETS) exposure on birth weight. A survey was conducted from 2000 to 2001 among 266 pregnant women who were hospitalized for delivery and on their singleton live births. We determined maternal GSTM1 and GSTT1 polymorphisms by polymerase chain reaction and measured the urinary cotinine of pregnant women at delivery by radioimmunoassay. Birth weight was found to decrease significantly with increasing concentrations of maternal urinary cotinine (P < 0.05). The interactive effect of exposure to ETS and the presence of the GSTT1 polymorphism was found to be significant by multivariate analysis (P < 0.01), whereas the interactive effect of exposure to ETS and the presence of GSTM1 polymorphism did not reach statistical significance (P = 0.21). A combination of the GSTM1-null and the GSTT1 null-genotypes was found to exacerbate the effect of maternal exposure to ETS on birth weight more than the presence of either genotype alone. Our data indicate that maternal exposure to ETS negatively affects neonatal birth weight, and the adverse effect of maternal exposure to ETS on neonatal birth weight could be modified by the maternal metabolic genotypes, GSTM1 and GSTT1.

Over-expression of thymosin Beta4 promotes abnormal tooth development and stimulation of hair growth

Thymosin beta 4 has multi-functional roles in cell physiology. It accelerates wound healing, hair growth and angiogenesis, and increases laminin-5 expression in corneal epithelium. Furthermore, thymosin beta 4 stimulates tumor growth and metastasis by induction of cell migration and vascular endothelial growth factor-mediated angiogenesis. Using a construct on the skin-specific keratin-5 promoter, we have developed thymosin beta 4 over-expressing transgenic mice to further study its functional roles. Thymosin beta 4 in adult skin and in embryonic stages of the transgenic mouse was analyzed by both Western blot and immunohistochemistry. The over-expression of thymosin beta 4 was observed especially around hair follicles and in the teeth in the transgenic mice. We examined the phenotype of the thymosin beta 4 over-expressing mice. Hair growth was accelerated. In addition, the transgenic mice had abnormally-shaped white teeth and dull incisors. We found that the expression of laminin-5 was up-regulated in the skin of the transgenic mice. We conclude that thymosin beta 4 has an important physiological role in hair growth and in tooth development.

Haplotype analysis of the matrix metalloproteinase-9 gene associated with advanced-stage endometriosis

OBJECTIVE To investigate whether the -1562C>T, R279Q, P574R, and R668Q polymorphisms of the matrix metalloproteinase-9 (MMP-9) gene are related to endometriosis. DESIGN Case-control study. SETTING University-based hospital in Korea. PATIENT(S) Patients with endometriosis stage III/IV (n = 225) who underwent pelvic surgery and controls (n = 198) with no endometriosis in a Korean population. INTERVENTION(S) Peripheral blood samples were collected by venipuncture. MAIN OUTCOME MEASURE(S) Frequencies of genotypes and haplotypes were compared with the risk of endometriosis including -1562C>T, R279Q, P574R, and R668Q polymorphisms of MMP-9. RESULT(S) In the two-locus haplotype analyses using the four single nucleotide polymorphisms (SNPs), an increase in the distribution of the R279Q/P574R (2678G>A/4859C>G) (AC haplotype: odds ratio [OR] = 3.180, 95% confidence interval [CI] = 1.956-5.170; GG haplotype: OR = 4.374, 95% CI = 2.376-8.053) and -1562C>T/R668Q (-1562C>T/5546G>A) (CA haplotype: OR = 3.280, 95% CI = 1.406-7.653) haplotypes was significantly associated with endometriosis. By contrast, the risk of endometriosis was not associated with the individual SNPs studied. CONCLUSION(S) These findings suggest that haplotype analysis was more informative than SNP analysis. The haplotypes in the MMP-9 gene may correlate with the progression of endometriosis, and further study of these variations might improve our understanding of the pathogenesis of endometriosis.

Immunohistochemical and quantitative competitive PCR analyses of midkine and pleiotrophin expression in cervical cancer

OBJECTIVE The aim of this study was to determine midkine (MK) and pleiotrophin (PTN) expression in cervical cancer. METHODS Prospective study in tertiary teaching hospital. Normal and cancerous cervical tissues were obtained from healthy women (n = 19) and from patients with cervical cancer (n = 42). The expressions of MK and PTN mRNA and protein were examined by quantitative competitive PCR and by immunohistochemistry. MK and PTN mRNA and protein expressions were examined with respect to tumor stage and size. RESULTS The expressions of midkine and pleiotrophin mRNA in cervical cancer were higher than those in the normal cervix (MK, 175.59 +/- 63.3 vs 1.00 +/- 0.18 fmol, respectively; PTN, 3.18 +/- 1.25 vs. 0.86 +/- 0.12 fmol, respectively, P < 0.05), and their expressions were not correlated with cervical cancer stage or size of the tumor. The expressions of MK and PTN protein in cancerous tissue were higher than those in the normal cervix (P < 0.05). Moreover, the protein expression of MK, but not of PTN, correlated with tumor stage and size. The expressions of MK and PTN were not correlated with vascular density. CONCLUSIONS Our results suggest that increased midkine mRNA and protein expressions are associated with the carcinogenesis of cervical cancer.

Polymorphisms and haplotypes of the gene encoding the estrogen-metabolizing CYP19 gene in Korean women: no association with advanced-stage endometriosis

AbstractA variety of factors affect the development of endometriosis, including hormonal status and genetic factors. The growth of endometriosis is stimulated by local estrogen production in conjunction with circulating estrogen. The CYP19 gene encodes a steroid aromatase that catalyses the conversion of C-19 androgens to estrogens. This study investigated whether polymorphisms of the CYP19 gene are associated with the risk of advanced endometriosis in Korean women. Blood samples were collected from 224 female patients with endometriosis of stages III and IV, as diagnosed by both pathologic and laparoscopic findings, and from a control group comprising of 188 women undergoing laparoscopic surgery or laparotomy for nonmalignant lesions. Single-nucleotide polymorphisms, restriction fragment length polymorphisms, and tetranucleotide tandem repeat polymorphisms were discriminated by the polymerase chain reaction (PCR). Haplotype analysis was also performed. CYP19 115T>C, 240G>A, and 1531C>T polymorphisms and [TTTA]n tetranucleotide repeat polymorphisms in the CYP19 gene and their haplotypes were not significantly associated with the risk of endometriosis. The risk of endometriosis also did not increase significantly with the number of higher risk alleles of the CYP19 gene. In conclusion, our findings suggest that CYP19 genetic polymorphisms are not associated with advanced-stage endometriosis in Korean women.

Single nucleotide polymorphisms and haplotypes of the genes encoding the CYP1B1 in Korean women: no association with advanced endometriosis.

AbstractObjective: To investigate whether single nucleotide polymorphisms and its haplotypes of gene encoding CYP1B1 are associated with the risk of advanced endometriosis in Korean women. Methods: We investigated 221 patients with histopathologically confirmed endometriosis rAFS stage III/IV and 188 control group women who were surgically proven to have no endometriosis. The genetic distribution of four different CYP1B1 polymorphisms at Ala119Ser, Leu432Val, Asp449(C>T), Asn453Ser were analyzed by polymerase chain reaction (PCR) and restriction fragment length polymorphism of PCR products. Haplotype analysis was also performed. Results: We found no overall association between each individual CYP1B1 genotype or haplotype and the risk of endometriosis. Also, the odds ratio of each haplotypes of CYP1B1 showed no association with the risk of endometriosis. Conclusions: These results suggest that CYP1B1 genetic polymorphism may not be associated with development of advanced endometriosis in Korean women.

Acute promyelocytic leukemia with i(17)(q10) on G-banding and PML/RARA rearrangement by RT-PCR without evidence of PML/RARA rearrangement on FISH.

The cytogenetic hallmark of acute promyelocytic leukemia (APL) is t(15;17)(q22;q21) involving the PML gene on chromosome 15q22 and the RARA gene on chromosome 17q21. Fluorescence in situ hybridization (FISH) can detect almost all PML/RARA rearrangements including complex and cryptic rearrangements [Kohno et al., 2001; Zaccaria et al., 2002; Brockman et al., 2003; Fujita et al., 2003]. Here, we describe a case of APL with the karyotype 46,XX,i(17)(q10)[12]/46,XX[8] without evidence of PML/ RARA rearrangement by FISH, but PML/RARA fusion gene was molecularly confirmed by RT-PCR. To our knowledge, this is the first report of APL with i(17)(q10), lacking evidence of PML/RARA rearrangement on FISH. A 44-year-old female was transferred to our hospital because of presence of some immature cells in peripheral blood. The laboratory findings were as follows: leukocyte count was 1.5 · 10/l, haemoglobin 12.6 g/dl and platelet count 49 · 10/l. On the peripheral blood smear, 5% immature cells were found. The bone marrow aspirate was replaced by hypergranular promyelocytes (82%) with multiple Auer rods. Cytochemistry studies demonstrated that myeloperoxidase, sudan black B and specific esterase were all positive. The bone marrow biopsy showed 80% cellularity. Flow cytometry studies showed that the immature cells were positive for CD13 and CD33 and negative for HLA-DR and CD34. The patient was diagnosed as having APL. For cytogenetic analysis, unstimulated short-term cultures were set up using the bone marrow aspirate. The cytogenetic analysis of bone marrow showed 46,XX,i(17)(q10)[12]/46,XX[8] in the 20 metaphase cells examined (Figure 1). FISH study was performed using PML/RARA dual color, dual fusion probe (32-191013,Vysis, Des Plaines, IL) according to the manufacturer’s instructions. The FISH signals were viewed using a ·100 oil immersion objective along with a dual-pass filter or single-pass filter and analysed independently in a blinded fashion by two persons. Each person analysed 200 nuclei from different areas of the same slide, for a total of 400 nuclei. When a t(15;17) is present, the nuclei typically show two yellow fusion signals: one for PML/RARA fusion on derivative chromosome 15 and one for RARAPML fusion on derivative chromosome 17, along with one red signal and one green signal, reflecting the PML and RARA signals on normal chromosomes 15 and 17, respectively. In the present case, FISH analysis showed two red and three green signals in 55% of interphase cells. Two red signals (PML gene) indicated both normal chromosome 15 and three green signals (RARA gene) were present: one for normal chromosome 17 and two for i(17)(q10) (Figure 2). No yellow fusion signals indicating PML/RARA rearrangement were detected. In addition, no other RARA FISH signals could be detected that might indicate a complex translocation or RARA translocation not involving PML. To identify PML/RARA rearrangement, RT-PCR was performed using bone marrow aspirate. The molecular study showed L-form PML/RARA chimeric transcript. She was treated with combination chemotherapy (idarubicin and Ara-C) and all trans retinoic acid. At the end of induction therapy, her bone marrow aspirate revealed complete remission status, but cytogenetic study showed 10% of metaphase cells bearing i(17)(q10) in 20 metaphase cells examined. The interphase FISH analysis showed three RARA signals in 20% of interphase cells, indicating persistent i(17)(q10). The PML/RARA rearrangement by RT-PCR was still positive. After consolidation chemotherapy, a complete haematological remission was achieved and no evidence of PML/RARA rearrangement was found on cytogenetic study, FISH or RT-PCR. FISH has been a useful tool for detection of PML/ RARA rearrangements in APL including cases without apparent abnormalities of chromosome 15 or chromosome 17 by conventional cytogenetics [Kohno et al., 2001; Zaccaria et al., 2002; Brockman et al., 2003; Fujita et al., 2003; Grimwade et al.,1997]. One report demonstrated that the clinical sensitivity of a commercially available PML/RARA probe using two LETTER TO THE EDITOR INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY

Association between MMP-2 and TIMP-2 gene polymorphisms and advanced-stage endometriosis in Korean women.

To characterize the genetic variation across the MMP‐2 and TIMP‐2 gene with the risk of advanced‐stage endometriosis.

Magnetic resonance imaging and positron emission tomography of a giant multiseptated pyomyoma simulating an ovarian cancer

A case of giant pyomyoma misdiagnosed as an ovarian cancer in a magnetic resonance imaging (MRI) and a positron emission tomography (PET) scan is presented. An unusual huge, multiseptated, and pedunculated character, as well as a normal-sized uterus on imaging studies, caused the misinterpretation of the MRI and PET scan images as an ovarian mucinous cystadenocarcinoma.