Aromatase deficiency caused by mutation of CYP19A1 gene: A case report / 中南大学学报(医学版)

Aromatase deficiency (AD) is a rare autosomal recessive genetic disease caused by loss-of-function mutations in aromatase gene (CYP19A1), leading to congenital estrogen deficiency syndrome. Both mothers of AD patients during pregnancy and female AD fetus show virilization, while male patients are usually diagnosed in adulthood due to continued height increase and metabolic abnormalities. In 2019, a patient with AD was admitted in the Second Xiangya Hospital. The patient was a 37-year-old adult male who continued to grow linearly after adulthood. His estradiol was below the measurable line, the follicle-stimulating hormone (FSH) increased, bone age delayed, epiphysis unfused, and the bone mass reduced. CYP19A1 gene detection showed that c.1093C>T, p.R365W was homozygous mutation. This disease is rare in clinic. Clinicians need to raise awareness of the disease for early diagnosis and treatment to improve the long-term prognosis of patients.

Influence of CYP19A1 gene expression levels in women with breast cancer: a systematic review of the literature

Breast cancer is the most frequently diagnosed malignant neoplasm in women and is considered a multifactorial disease of unknown etiology. One of the major risk factors is genetic alteration. Changes in CYP19A1 gene expression levels have been associated with increased risk and increased aggressiveness of breast cancer. Increased CYP19A1 gene expression and/or aromatase activity are among the major regulatory events for intratumoral production of estrogens in breast malignant tissues. This systematic review aimed to investigate the influence of CYP19A1 gene expression levels in women with breast cancer. The research was carried out using the PubMed, Scopus, and Web of Science databases. Searches were conducted between February 2 and May 15, 2019. Inclusion criteria were studies published between 2009 and 2019, English language publications, and human studies addressing the gene expression of CYP19A1 in breast cancer. A total of 6.068 studies were identified through PubMed (n=773), Scopus (n=2,927), and the Web of Science (n=2,368). After selecting and applying the inclusion and exclusion criteria, six articles were included in this systematic review. This systematic review provides evidence that increased or decreased levels of CYP19A1 gene expression may be related to pathological clinical factors of disease, MFS, OS, DFS, WATi, markers of metabolic function, concentrations of E1, FSH, and in the use of multiple exons 1 of the CYP19A1 gene in breast cancer.

Loss-of-function of sox3 causes follicle development retardation and reduces fecundity in zebrafish

Folliculogenesis is essential for production of female gametes in vertebrates. However, the molecular mechanisms underlying follicle development, particularly apoptosis regulation in ovary, remain elusive. Here, we generated sox3 knockout zebrafish lines using CRISPR/Cas9. sox3 knockout led to follicle development retardation and a reduced fecundity in females. Comparative analysis of transcriptome between sox3 and wild-type ovaries revealed that Sox3 was involved in pathways of ovarian steroidogenesis and apoptosis. Knockout of sox3 promoted follicle apoptosis and obvious apoptosis signals were detected in somatic cells of stages III and IV follicles of sox3 ovaries. Moreover, Sox3 can bind to and activate the promoter of cyp19a1a. Up-regulation of Cyp19a1a expression promoted 17β-estradiol synthesis, which inhibited apoptosis in follicle development. Thus, Sox3 functions as a regulator of Cyp19a1a expression, via 17β-E2 linking apoptosis suppression, which is implicated in improving female fecundity.

Pretentious genomic selection signatures in CYP19A1 gene associated with silent estrous behavior in water buffalo in Pakistan

Background: Poor reproductive efficiency of river buffalos hampers the production capabilities of animals. Buffalos are mainly considered poor breeders owing to the constrained expression of estrus behavior. Failure to display heat signs is an indication of improper functionality of signaling peptides to trigger on a series of behavioral changes, which can be detectable by breeders for timely insemination of females. This might cause an animal to be a repeat breeder. Genomic variations underlying synthesis of signaling peptides can be a useful marker to select superior animals with better reproductive efficiency. In this context, the current study was designed to analyze the CYP19A1 gene in Nili-Ravi buffalo. Results: A total of 97 animals were selected and were divided into two groups on the basis of their heat score. PCR amplification and sequencing of the amplicons were performed using the specific sets of primer, and then, sequences were analyzed for novel variants. A total of 11 polymorphic sites were identified illustrating phenotypic variation in the heat score. Most of the loci were found homologous. Single Nucleotide Polymorphisms (SNPs) were analyzed for association with silent estrus. A three-dimensional protein model was also generated to locate the position of exonic SNPs. Conclusion: This study illustrated that polymorphic sites in the CYP19A1 gene provided potential markers for selection of buffalos with better estrus behavior.

Association of CYP19A1 gene rs7176005 single nucleotide polymorphism with breast cancer risk and clinicopathologic features of tumor / 中华预防医学杂志

Objective@#The aim of this study was to investigate the association of the CYP19A1 rs7176005 single nucleotide polymorphism (SNP) with breast cancer risk and with clinicopathologic features of tumors.@*Methods@#This study was conducted by including 138 patients with breast cancer (cancer group), those who diagnosed as primary breast cancer after operation by pathology. There were 293 cases in the group of benign breast disease which was presented as a solid mass by the color ultrasound and pathologically diagnosed as "fibroadenoma or adenosis" (benign breast disease group), the cases were paired with breast cancer patients by age±5 in the same period, and there were 259 cases in the group of healthy control who received routine physical examination during the same period and were paired with breast cancer patients by age±5 without any detection of breast related diseases (healthy control group) at West China hospital between September 2012 and November 2016. The CYP19A1 rs7176005 SNP was detected by a direct sequencing method. Hardy-Weinberg test was used to analyze the genetic balance of the 3 groups. Chi square test was used to compare the distribution of rs7176005 genotypes between the 3 groups, and the differences of clinicopathological features in breast cancer patients carrying different genotypes.@*Results@#The ages of the breast cancer cases, the benign breast disease group and the healthy control group were (44.69±8.09), (42.33±11.44) and (41.92±9.61) years old, respectively. Hardy-Weinberg equilibrium test identified that the composition ratios of alleles C and T in breast cancer group, benign breast disease group and healthy group were not statistically significant (χ2 values were 0.83, 0.34 and 0.04, respectively, P values were 0.363, 0.561, and 0.852, respectively). All the three groups met the genetic balance, had consistency and could represent the population. Among the 138 cases of breast cancer, the CYP19A1 rs7176005 SNP was significantly associated with the diameter of the tumor (P=0.031). The majority of tumor size was <2 cm in patients who carrying TT and CT genotypes, and the proportion was 75% (12/16) and 58% (40/69), respectively. While those patients with TT genotype were mainly >2 cm and ≤5 cm, and the proportion was 51% (27/53). The distribution of TNM stage among patients with different genotypes was also statistically significant (χ2=11.19, P=0.025). The most common stage was Ⅱ in Patients who carrying CC and CT genotypes, and the proportion was 45.3% (24/53) and 52.2% (36/69), respectively. While those patients with TT genotype was mainly in stage Ⅰ and the proportion was 56.3% (9/16).@*Conclusion@#Though the CYP19A1 rs7176005 SNP is not associated with breast cancer development, breast cancer patients with the C allele exhibit a high tumor growth rate and large diameters.

Effects of Si-Wu Mixture on Ovarian Granulose Cell Treated with Cisplatin through TGF-β3 Protein Pathway / 世界科学技术-中医药现代化

This study was aimed to investigate the effects of Si-Wu mixture on the secretion of E2 and the expression of CYP19al gene in granulosa cells after cell injury.Ovarian granulosa cells of SD rats were treated with cisplatin (CDDP).And the expression levels of E2 and CYP19a1 were determined by different testing methods.The results showed that the level of E2 induced by radioimmunoassay of the Si-Wu group was significantly higher than that of the CDDP group.Meanwhile,the group which added TGF-β3 protein pathway blocker was lower than others.The results of immunohistochemistry and western blotting showed that the expression level of CYP 19a1 of the Si-Wu group was higher than that of the CDDP group.In the western blotting,the group which added blocker was significantly lower than the non-blocking group.It was concluded that the pharmacological serum of Si-Wu mixture can enhance the level of E2 in CDDP cells through TGF-β3 protein pathway.And the effect is accomplished by the intervention of CYP 19a1.

Polymorphism C242T in the Cyp19 gene in meat sheep / Polimorfismo no gene cyp19 em ovinos de corte

Abstract The objective of this study was to evaluate the frequency of C242T polymorphism on the aromatase gene and the allelic and genotypic frequency of these variants in sheep belonging to four breed groups. Blood samples were collected from 187 animals of four breed groups: Dorper, Santa Inês, Texel and White Dorper, originated from herds in the region of Maringá/PR, Brazil. The genomic DNA was extracted using alkaline extraction, with subsequent amplification of the fragments via PCR with specific primer. The samples resulting from amplification were subjected to digestion process using the Dpn II restriction enzyme and to polyacrylamide gel electrophoresis 10.0% and stained with silver nitrate. Three distinct genotypes were observed: homozygous (CC), heterozygous (CT) and homozygous for no cut (TT). The resulting data were analyzed using the POPGENE software with 5% significance. Genotypic frequencies among the breed groups were: Texel (CC - 0.426; CT - 0.511; TT - 0.064), Dorper (CC - 0.073; CT - 0.732; TT - 0.439), White Dorper (CC - 0.021; CT - 0.255; TT - 0.723) and Santa Inês (CC - 0.115; CT - 0.462; TT - 0.423).

Resumo O objetivo deste trabalho foi avaliar as frequências alélicas e genotípicas do polimorfismo do C242T no gene da aromatase em ovinos pertencentes a quatro grupos raciais. Foram coletadas amostras de sangue de 187 animais de quatro grupos raciais: Dorper, Santa Inês, Texel e White Dorper, provenientes de rebanhos da região de Maringá, PR - BR. O DNA genômico foi extraído utilizando o método de extração alcalina, com posterior amplificação dos fragmentos via PCR com primer específico. As amostras resultantes da amplificação foram submetidas ao processo de digestão com auxilio da enzima restrição Dpn II e submetido à eletroforese em gel de poliacrilamida de 10,0% e corado nitrato de prata. Foram observados três genótipos distintos: Homozigoto (CC), heterozigoto (CT) e homozigoto para não corte (TT). Os dados resultantes foram analisados utilizando o software POPGENE com significância de 5%. As frequências genotípicas entre os grupos raciais foram: Texel (CC - 0,426; CT - 0,511; TT - 0,064), Dorper (CC - 0,073; CT - 0,732; TT - 0,439), White Dorper (CC - 0,021; CT - 0,255; TT - 0,723) e Santa Inês (CC - 0,115; CT - 0,462; TT - 0,423).

Effects ofJingui Shenqi Pills on Expressions of Testicular TGF-β1 and CYP19 in Kidney-yang-deficiency Rats / 中国中医药信息杂志

Objective To observe the effects ofJingui Shenqi Pills on the expressions of TGF-β1, CYP19, sex hormone level and sperm quality;To discuss its mechanism for the treatment of male sterility.Methods Forty adult male SD rats were randomly divided into normal group, model group, retardant group andJingui Shenqi Pills group, 10 rats in each group. Except for the normal group, adenine was used to induce healthy rats to kidney yang deficiency and sterility model rats. Normal group was given normal saline for gavage;retardant group was injected with testicular mesenchyme TGF-β1;Jingui Shenqi Pills group was givenJingui Shenqi Decoction for gavage. Blood was taken through caudal vein. Immunohistochemical, radioimmunoassay and electron microscopy observation were used to detect the expressions of TGF-β1, CYP19, sex hormone level, sperm density and motility rate and testicular morphological changes.Results Compared with normal group, the expression of TGF-β1 in the model group increased and CYP19 decreased (P<0.05). Compared with model group, the expression of TGF-β1 decreased and the expression of CYP19 increased (P<0.05). Compared with normal group, the number of sperm quantity was small and paramorphia in model group (P<0.05). Compared with model group, T, E2, sperm density and motility rate increased significantly inJingui Shenqi Pills group and retardant group (P<0.05). There was no statistical significance among indexes in retardant group andJingui Shenqi Pills group.ConclusionJingui Shenqi Pills can improve sperm quality and sex hormone level of model rats with kidney-yang-deficiency, which mechanism is probably realized by inhibiting the expression of TGF-β1R and promoting the expression of CYP19 in testis to effect development and proliferation of sperm.

Aromatase activity in brain and ovary: Seasonal variations correlated with circannual gonadal cycle in the catfish, Heteropneustes fossilis.

Seasonal variations in the aromatase activity in H. fossilis estimated by a microassay were correlated with the sex steroids, vitellogenin in and ovarian weight during circannual reproductive cycle. In the female catfish, aromatase activity was detectable in the hypothalamus throughout the year whereas in ovary only during active vitellogenesis. In the catfish, hypothalamic aromatase levels increased two times during annual gonadal cycle, once in a fully gravid fish and then in a reproductively quiescent fish. On the other hand, increase in the ovarian aromatase activity was observed only during vitellogenesis, which showed a direct correlation with plasma levels of sex steroids. Further, plasma levels of testosterone and estradiol suggested a precursor-product relationship. At the completion of vitellogenesis, ovarian aromatase activity declined sharply resulting in elevation of plasma testosterone levels, which in turn could be utilized as substrate by the hypothalamic aromatase whose activity was the highest in the postvitellogenic catfish. At least two isoforms of gene, cyp19a and cyp19b, coding for aromatase in ovary and brain respectively were expressed in the catfish. Aromatase activity was more concentrated in those areas of catfish brain, which have been implicated in the control of reproduction.

CYP17, CYP19, and NQO1 Genetic Polymorphisms and Breast Cancer Susceptibility in Young Women in Brazil.

Aims: Breast cancer is the most common cancer in women worldwide, being exposure to estrogens the acknowledged main risk factor. CYP17, CYP19 and NQO1 are enzymes involved in the estrogen metabolism, so their polymorphisms may be involved in breast carcinogenesis. The aim of this study was to determine the magnitude of the association between CYP17 MspA1, CYP19 Arg264Cys, and NQO1 C609T polymorphisms and breast cancer in young women. Methods: This is a hospital-based case-control study carried out in Rio de Janeiro. Cases were 270 women with age range 18-35 years and a histopathological diagnosis of breast cancer between 1999-2009. Controls were 270 women without cancer at the same age range. Results: An association between CYP17 MspA1 or CYP19 Arg264Cys polymorphisms and breast cancer were not observed (OR = 1.02, 95% CI 0.72-1.44 for CYP17 genotypes TC/CC and OR = 0.85, 95% CI 0.48-1.49 for CYP19 genotypes CT/TT). However, a statistically significant increased risk estimate was identified in women who had at least one NQO1 polymorphic allele (T), OR= 1.96, 95% CI 1.13-3.40 following adjustment for selected confounders. Conclusion: This study suggests that the NQO1 609T polymorphism may be a risk factor for breast carcinogenesis in women less than 36 years in Brazil.

CYP19 A1 gene polymorphism and polycystic ovarian syndrome / 医学研究生学报

Objective The CYP19 gene product enzyme aromatase mediates the conversion of the androgen testosterone to es -tradiol.The aim of this study is to investigate whether the CYP19A1 gene single nucleotide polymorphism (SNP) is associated with the susceptibility to polycystic ovary syndrome (PCOS) and serum hormone levels. Methods We conducted a case-controlled study, which included 373 PCOS patients and 313 healthy controls.We genotyped SNP rs2899470 in the subjects using the polymerase chain re-action-restriction fragment length polymorphism ( PCR-RFLP) method and analyzed the frequencies of genotypes and alleles as well as the association of different genotypes with age , menarchal age, body mass index (BMI), and serum levels of hormones. Results The gen-otypic distributions of rs2899470 GG, TG, and TT in the PCOS women were 44.5%, 49.6%, and 5.9%, respectively, significantly dif-ferent from 39.3%, 48.6%, and 12.1%in the healthy controls (P=0.013).The frequency of the G allele was 69.3%in the former, remarkably higher than 63.6%in the latter (P=0.025).The rs2899470 genotypic frequencies were associated with the serum E 2/T lev-els in the PCOS patients. Conclusion SNP rs2899470 in the CYP19A1 gene is associated with the susceptibility to PCOS , and so is the genotype of rs2899470 with serum E2/T levels, which may be attributed mainly to the reduced activity of aromatase .

CYP19 gene variant confers susceptibility to endometriosis-associated infertility in Chinese women

An aromatase encoded by the CYP19 gene catalyzes the final step in the biosynthesis of estrogens, which is related to endometriosis development. To assess the association of CYP19 gene polymorphisms with the risks of endometriosis, chocolate cysts and endometriosis-related infertility, a case-control study was conducted in Chinese Han women by recruiting 225 healthy control females, 146 patients with endometriosis, 94 endometriosis women with chocolate cyst and 65 women with infertility resulting from endometriosis, as diagnosed by both pathological and laparoscopic findings. Individual genotypes at rs2236722:T>C, rs700518:A>G, rs10046:T>C and [TTTA]n polymorphisms were identified. Allelic and genotypic frequencies were compared between the control group and case groups by chi-square analysis. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were determined by logistic regression analysis to predict the association of CYP19 gene polymorphisms with the risk of endometriosis, the related chocolate cysts and infertility. The genotype distributions of the tested CYP19 gene polymorphisms were not significantly different between the healthy control group and the endometriosis/endometriosis with the chocolate cyst group. However, the CYP19 rs700518AA genotype was significantly associated with an increased risk of endometriosis-related infertility (55.4% in the infertility group vs 25.3% in the control group, P<0.001; OR (95% CI): 3.66 (2.06-6.50)) under the recessive form of the A allele. Therefore, we concluded that in Chinese Han females CYP19 gene polymorphisms are not associated with susceptibility to endometriosis or chocolate cysts, whereas CYP19 rs700518AA genotype confers genetic susceptibility to endometriosis-related infertility.

Molecular mechanisms of luteinization

Dynamic changes in steroidogenesis occur in ovarian granulosa cells during ovulation after the LH surge. The ovulatory LH surge induces rapid up-regulation of steroidogenic acute regulatory (StAR) protein and rapid down-regulation of aromatase (Cyp19a1) in granulosa cells undergoing luteinization during ovulation. These rapid changes in StAR and Cyp19a1 gene expression after the LH surge efficiently facilitate progesterone production, which plays a crucial role in ovulation and the following luteinization. Recently, it has become clear that epigenetic regulation such as histone modifications and DNA methylation play a key role in gene expression through the chromatin remodeling of the promoter region. This study reports the in vivo evidence that epigenetic mechanisms including histone modifications, DNA methylation and chromatin remodeling are involved in the rapid changes of StAR and Cyp19a1 gene expression in granulosa cells undergoing luteinization during ovulation.

Mulheres inférteis com endometriose pélvica mínima e leve submetidas à estimulacão ovariana apresentam menor expressão do gene CYP19A1 em células do cumulus / Infertile women with minimal and mild endometriosis submitted tocontrolled ovarian stimulation present lower expression of the gene CYP19A1 in cumulus cells

Objetivo: Menores taxas de gestação em portadoras de endometriose submetidas a técnicas de reprodução assistida podem estar relacionadas à piora da qualidade oocitária. A análise da expressão gênica em células do cumulus (CC) pode fornecer biomarcadores passíveis de predizer a qualidade gamética. O objetivo deste estudo foi comparar os níveis da expressão do gene CYP19A1 em CC de mulheres inférteis com endometriose mínima/leve (I/II) e controles inférteis. Método: Foram selecionadas pacientes com infertilidade por endometriose pélvica inicial e por fator masculino e/ou tubário (grupo controle), submetidas à estimulação ovariana controlada para injeção intracitoplasmática de espermatozoide (ICSI). Imediatamente após a captação oocitária, CC foram isoladas e armazenadas. Foi realizada a quantificação da expressão do gene CYP19A1 nas CC por meio de PCR-Real Time. Resultados: Foram isoladas CC de 23 mulheres inférteis com endometriose I/II e de 41 controles. Observou-se expressão significativamente menor do gene CYP19A1 em CC de mulheres inférteis com endometriose I/II (0,56 ± 0,17) quando comparadas às controles (0,15 ± 0,04) (p = 0,043). Conclusões: A menor expressão do gene CYP19A1 em CC de mulheres inférteis com endometriose pélvica em estágios iniciais pode mediar a piora da qualidade oocitária, abrindo novas perspectivas no entendimento da etiopatogênese da infertilidade relacionada à doença.

Objective: Lower pregnancy rate in women with endometriosis submitted to assisted reproductive techniques might be related to poor oocyte quality. The analysis of the expression of the genes in cumulus cells (CC) might provide biomarkers that can predict gamete quality. The main objective of the present study was to compare the levels of the expression of the gene CYP19A1 in CC of infertile women with minimal and mild (I/II) endometriosis and infertile controls. Method: There were selected patients with infertility caused by initial pelvic endometriosis and by male/tubal factor (control group), submitted to controlled ovarian stimulation to ICSI. Immediately after the oocyte retrieval, CC were isolated and stored. Quantification of the expression of the gene CYP19A1 in CC was performed using PCR-real time.Results: CC were isolated from 23 infertile women with endometriosis I/II and 41 from control. Significant lower expression of the gene CYP19A1 in CC was observed in infertile women with endometriosis I/II (0.56 ± 0.17) when compared to control (0.15 ± 0.04) (p = 0,043). Conclusions: The lower expression of the gene CYP19A1 in CC of infertile women with pelvic endometriosis in initial stages might mediate the poor oocyte quality, opening new perspectives on the understanding of the etiopathogenesis of infertility related to the disease.

Correlation analysis of cytochrome P450c19 genes with osteoporotic fracture in a large sample of Shaanxi Guanzhong Population / 西安交通大学学报(医学版)

Objective To study the relationship between osteoporotic hip fracture (OHF) and polymorphism of cytochrome P450c19 genes. Methods By using a group correlation analysis, we made a case-control study of seven single nucleotide polymorphism (SNP) sites groupinge and haplotype in 400 OHF patients from Shaanxi Guanzhong population and 400 normal samples as controls. Results A total of 700 samples were successfully genotyped. Correlation analysis identified three blocks and selected seven tag SNPs. According to the variance analysis, rs7167343, rs8031463, and polymorphism of haplotype 3 were associated with hip fracture. Conclusion Cytochrome P450c19 genes have association with the incidence of osteoporotic fracture in the Han population we studied from Shaanxi Guanzhong region. Therefore, cytochrome P450c19 genes may affect the incidence of osteoporotic hip fracture in the Han population from Guanzhong region, Shaanxi Province.

Estudio de análogos de la sulfonanilida que actúan como supresores de la expresión de la aromatasa / Study of sulfonanilide analogues acting as suppressants of the aromatase expression

Se realizó un estudio tridimensional cuantitativo de relación-estructura con nuevos análogos de la sulfonanilida que actúan como supresores de la expresión y actividad de la aromatasa en células de cáncer de mama, independientemente de la inhibición de la ciclooxigenasa-2 (COX-2). La superposición molecular de los ligandos en la plantilla fue llevada a cabo por el método Database Alignment. El mejor modelo estuvo constituido por la combinación de los campos estérico e hidrofóbico, los cuales arrojaron los siguientes parámetros: q² = 0,613 con siete componentes, R² no validado igual a 0,976, valor de F igual a 81,695; 0,347 y 0,047 para los errores estándar de predicción y estimación, respectivamente. Las contribuciones estérica e hidrofóbica fueron de 50,5 y 49,5% respectivamente. Los datos generados por el presente estudio podrían impulsar el diseño de nuevos y más potentes reguladores selectivos de la expresión de la aromatasa.

A three-dimensional quantitative structure-activity relationship study was performed with new Sulfonanilide analogue molecules acting as aromatase expression and activity suppressors in breast cancer cells independent of COX-2 inhibition. The molecular supression of the ligands in the grid was carried out by the DATA-BASE ALIGNMENT method. The best model formed by combining the esteric fields and hydrophobic fields yielded the following parameters: q² = 0.613 with seven components, not validated R² equal to 0.976, with an F value of 81.695, 0.347 and 0.047 for the standard errors of prediction and estimate, respectively. The contribution of esteric and hydrophobic fields was 50.5 and 49.5% respectively. data generated from this study may be used to design new and more potent selective aromatase expression regulators.

El splicing alternativo del exón 5 de la citocromo p450 aromatasa podría ser un mecanismo de regulación de la producción de estrógenos en humanos / Exon 5 alternative splicing of the cytochrome P450 aromatase could be a regulatory mechanism for estrogen production in humans

La enzima P450 aromatasa (P450Aro) participa en la síntesis de estrógenos a partir de andrógenos. La mutación c655G>A, descripta en forma heterocigota en una niña y en forma homocigota en un hombre adulto, ambos con déficit de aromatasa, genera la disrupción del sitio dador de splicing exón5-intrón5. Se ha postulado que la retención del intrón5 y la generación de una proteína truncada inactiva serían las consecuencias de esta mutación. Sorpresivamente, la paciente presentó desarrollo espontáneo de mamas y niveles puberales de estradiol, sugiriendo una actividad aromatasa (AA) residual. En principio postulamos que la mutación c655G>A generaría la pérdida del exón5 con conservación del marco de lectura, generándose una proteína con menor actividad que podría explicar el déficit parcial. La expresión del ARNm sin exón5 (ARNm- E5) en linfocitos de la paciente sugiere una asociación entre la pérdida del exón y la presencia de la mutación; posteriormente confirmada realizando ensayos de splicing en células Y1. Sin embargo, la expresión del cDNAE5 en células Y1 presentó una AA nula que no explicaría un déficit parcial. La expresión del ARNm-E5 fue detectada en placenta, testículo y adrenal humanos como una variante de splicing normal. Estos resultados indicarían la ocurrencia de splicing alternativo (SA) en la zona codificante de P450Aro como un posible mecanismo regulador de la producción de estrógenos en tejidos esteroidogénicos humanos. La mutación c655G>A podría alterar los mecanismos fisiológicos reguladores del SA del exón5 favoreciendo su exclusión. De esta forma, bajos niveles de ARNm+E5 podrían expresarse aun en presencia de la mutación explicando el fenotipo de déficit parcial observado en la paciente.

P450 aromatase (P450Aro), involved in androgen to estrogen conversion, is encoded by the CYP19 gene. P450Aro c655G>A mutation described in heterozygous form in a girl and in homozygous form in an adult male with P450Aro deficiency results in an aberrant splicing due to disruption of a donor splice site. A truncated inactive protein would be expected if intron5 is retained. Surprisingly, the girl described with this mutation showed spontaneous breast development and pubertal estradiol (E2) levels suggesting residual P450Aro activity (AA). Formerly, we postulate the in frame E5 skipping as a consequence of this mutation generating a protein with some degree of activity. When P450Aro mRNA expression was analysed from patient's lymphocytes, an aberrant spliced mRNA lacking E5 (-E5mRNA) was detected, suggesting an association between E5 skipping and the presence of the mutation. Splicing assays in Y1 cells confirmed this association. -Ex5 cDNA expression in Y1 cells resulted in an inactive protein that could not explain patient's phenotype. Exon 5 might be predicted as a poorly defined exon suggesting a susceptibility to splicing mutations and physiological alternative splicing (AS) events. Therefore, -Ex5mRNA was assessed as a natural occurring alternative transcript in normal human steroidogenic tissues. As P450Aro -E5mRNA expression was detected in human term placenta, prepubertal testis and prepubertal adrenal, we might speculate that AS of P450Aro coding region would occur in humans and would be involved in the complex AA regulation. Furthermore, tissue specific regulation of AS might suggest low expression of +E5mRNA from the c655G>A allele explaining residual AA evidenced in the affected girl.

No association of Polymorphism in the Estrogen-synthesizing Enzyme Genes CYP19 with Advanced Endometriosis in Korean Women / 대한불임학회지

OBJECTIVE: To investigate whether polymorphism of CYP19 gene is associated with the risk of advanced endometriosis in Korean women. METHODS: Blood samples were collected from 202 endometriosis patients and 221 controls. The patients with endometriosis of stages III and IV diagnosed by both pathologic and laparoscopic findings to according modified AFS classification. The women undergoing laparoscopic surgery or laparotomy for non-malignant lesions were included in the control group. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of PCR products were done to determine all individuals' genotype. RESULTS: The heterozygous allele in CYP19 gene was the most common genotypes in both endometriosis and healthy control groups (52.0% vs. 46.1%). CYP19 gene polymorphisms did not show the significant differences between the control group and endometriosis group. CONCLUSION: The results suggested that the CYP19 genetic polymorphism was not associated with a risk of advanced endometriosis in Korean women.

CYP19 Gene Polymorphism in Patients with Endometriosis / 대한산부인과학회지

OBJECTIVE: To explore the association of the CYP19 gene polymorphism with the risk of endometriosis. METHODS: Two hundred seventy-nine women with surgically or histologically diagnosed endometriosis of stages I-IV (ASRM, 1997) were recruited, and two hundred eighteen patients with no evidence of endometriosis by laparoscopy or laparotomy served as control. We analysed the frequency and distribution of a TTTA repeat polymorphism and a 3 bp insertion (I)/deletion (D) polymorphism in intron 4 of the CYP19 gene. RESULTS: Six alleles of the CYP19 gene tetranucleotide repeat polymorphism were found in subjects: from 7 repeats to 13 repeats except 9 repeats. There was no statistically significant difference in the allele distribution of tetranucleotide repeat polymorphism in intron 4 of the CYP19 gene between patients with endometriosis and controls. Also there was no statistically significant difference in the 3 bp insertion (I)/deletion (D) polymorphism in intron 4 of the CYP19 gene between patients with endometriosis and controls. CONCLUSION: These results suggest that tetranucleotide repeat polymorphism and a 3 bp insertion (I)/deletion (D) polymorphism of the CYP19 gene are not associated with the risk endometriosis in the Korean population.