ABSTRACT
This study aimed to define the expression patterns of HENMT1 and PIWI proteins in human testis and investigate their association with transposon expression, infertility sub-type or development of testicular germ cell tumours (TGCTs). Testis biopsies showing normal spermatogenesis were used to identify normal localisation patterns of HENMT1 and PIWIL1 by immunolocalisation and RT-PCR after laser microdissection. 222 testis biopsies representing normal spermatogenesis, hypospermatogenesis, spermatogenic arrests, Sertoli cell-only (SCO) tumours and TGCTs were analysed by RT-qPCR for expression of HENMT1/PIWIL1/PIWIL2/PIWIL3/PIWIL4 and LINE-1 Additionally, HENMT1-overexpressing TCam2 seminoma cell lines were analysed for the same parameters by RT-qPCR. We found that HENMT1 and PIWIL1 are coexpressed in pachytene spermatocytes and spermatids. Expression of HENMT1, PIWIL1 and PIWIL2 was mainly dependent on germ cell content but low levels of expression were also detected in some SCO samples. Levels of HENMT1, PIWIL1 and PIWIL2 expression were low in TGCT. Samples with HENMT1, PIWIL2 and PIWIL4 expression showed significantly (P < 0.05) lower transposon expression compared to samples without expression in the same histological group. HENMT1-overexpressing TCam2 cells showed lower LINE-1 expression than empty vector-transfected control lines. Our findings support that the transposon-regulating function of the piRNA pathway found in the mouse is conserved in adult human testis. HENMT1 and PIWI proteins are expressed in a germ-cell-specific manner and required for transposon control.
Subject(s)
Argonaute Proteins/genetics , DNA Transposable Elements , Methyltransferases/genetics , Neoplasms, Germ Cell and Embryonal/genetics , Seminoma/genetics , Sertoli Cell Tumor/genetics , Sertoli Cell-Only Syndrome/genetics , Testicular Neoplasms/genetics , Testis/enzymology , Adolescent , Adult , Aged , Argonaute Proteins/metabolism , Cell Line, Tumor , Fertility/genetics , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Neoplastic , Humans , Long Interspersed Nucleotide Elements , Male , Methyltransferases/metabolism , Middle Aged , Neoplasms, Germ Cell and Embryonal/enzymology , Neoplasms, Germ Cell and Embryonal/pathology , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Seminoma/enzymology , Seminoma/pathology , Sertoli Cell Tumor/enzymology , Sertoli Cell Tumor/pathology , Sertoli Cell-Only Syndrome/enzymology , Sertoli Cell-Only Syndrome/physiopathology , Spermatogenesis/genetics , Testicular Neoplasms/enzymology , Testicular Neoplasms/pathology , Testis/pathology , Testis/physiopathology , Young AdultSubject(s)
Hemorrhage/pathology , Racemases and Epimerases/metabolism , Sertoli Cell Tumor/pathology , Testicular Neoplasms/pathology , Biomarkers, Tumor/metabolism , Hemorrhage/etiology , Humans , Male , Middle Aged , Orchiectomy , Sertoli Cell Tumor/complications , Sertoli Cell Tumor/enzymology , Testicular Neoplasms/complications , Testicular Neoplasms/enzymology , Testis/diagnostic imaging , Testis/pathology , Testis/surgery , UltrasonographyABSTRACT
Large cell calcifying Sertoli cell tumors (LCCSCT) are associated with Carney complex and Peutz-Jeghers syndrome. The mechanisms linking these 2 genetic defects to the genesis of this tumor are obscure. Studies of CYP19 (aromatase) and transforming growth factor (TGF)-beta1 messenger RNA (mRNA) abundance, estrogen receptor (ER), TGFbeta1, and TGFbeta type II receptor (R) immunochemistry were carried out in the testis of a patient with this tumor to gain information on possible mechanisms of cell tumor development. Testicular tissue of a prepubertal patient, collected at gonadectomy, was separated into 2 macroscopically distinct fractions: tumoral nodules (Tu) and extratumoral, normal-looking testicular tissue (ExTu). The patient was a 9.5-year-old boy with a 5-year history of bilateral gynecomastia (Tanner stage 4), no pubic hair, incipient genital development, and bilateral testicular nodules. Multiple pigmented lesions of the skin were present. Bilateral mammectomy and gonadectomy was performed. RNA was extracted from Tu and ExTu for semiquantitative reverse transcriptase-polymerase chain reaction of CYP19 and TGFbeta1. Protein expression of ER, TGFbeta1, and TGFbeta type II R in Tu and ExTu was detected by immunohistochemistry. Cell proliferation was estimated by Ki-67 antigen immunochemistry and apoptosis using a modified TUNEL assay. Mean expression of aromatase and TGFbeta1 mRNAs in Tu was 6- and 2.3-fold higher than in ExTu, respectively (P<0.05). Tumoral cells exhibited ER staining with a predominant extranuclear localization. Positive staining of Sertoli cells in Tu was higher than in ExTu. TGFbeta1 immunostaining of the interstitial cells in Tu was higher than in ExTu. TGFbeta type II R immunostaining was detected in most Sertoli and interstitial cells, but intensity in ExTu was lower than in Tu. No significant difference was detected in the proliferation index, but in Tu, the percentage of Sertoli cells in apoptosis (1.4%) was significantly lower (P<0.01) than in ExTu (14.0%). The following hypothesis is proposed. The congenital gene defects of Carney complex or of Peutz-Jeghers syndrome might trigger a cascade of intracellular events that leads to overexpression of aromatase in Sertoli cells, favoring the development of a LCCSCT. At some point in the evolution of the disease, a mutational event might induce a higher expression of the ER. Also, TGFbeta1 protein expression is increased in neighboring cells. In this environment, TGFbeta1 might switch from tumor suppressor to oncogenic factor and, along with estrogen-ER complexes, might favor tumor progression by inhibiting apoptosis.
Subject(s)
Aromatase/metabolism , Calcinosis , Receptors, Estrogen/metabolism , Sertoli Cell Tumor , Testicular Neoplasms , Transforming Growth Factor beta/metabolism , Apoptosis , Aromatase/genetics , Child , Gene Expression , Gynecomastia/surgery , Humans , Immunohistochemistry , Male , Models, Biological , Orchiectomy , RNA, Messenger/analysis , Receptors, Estrogen/genetics , Sertoli Cell Tumor/enzymology , Sertoli Cell Tumor/etiology , Sertoli Cell Tumor/genetics , Sertoli Cell Tumor/metabolism , Sertoli Cell Tumor/pathology , Testicular Neoplasms/enzymology , Testicular Neoplasms/etiology , Testicular Neoplasms/genetics , Testicular Neoplasms/metabolism , Testicular Neoplasms/pathology , Transforming Growth Factor beta/geneticsABSTRACT
Aromatase, 17 alpha-hydroxylase, and cholesterol side-chain cleavage P-450 cytochromes (P-450AROM, P-450(17 alpha,) and P-450SCC, respectively) were immunohistochemically localized in nine granulosa cell tumors, 15 thecomas, ten Sertoli-Leydig cell tumors, two steroid cell tumors, five fibromas, and five sclerosing stromal tumors. In the thecomas, P-450SCC and P-450(17 alpha) were positive in luteinized theca cells and in cells with vacuolated cytoplasm, while P-450AROM was not observed. In the steroid cell tumors, all the P-450 cytochromes were intensely stained. P-450SCC and P-450(17 alpha) were present in cells with vacuolated cytoplasm in two cases of sclerosing stromal tumor. P-450AROM was weakly demonstrated in one of the granulosa cell tumors. P-450(17 alpha,) P-450SCC, and P-450AROM were all faintly stained in the Sertoli-Leydig cell tumors. No P-450 cytochrome immunoreactivity was observed in any fibroma.
Subject(s)
Aromatase/metabolism , Cholesterol Side-Chain Cleavage Enzyme/metabolism , Ovarian Neoplasms/enzymology , Steroid 17-alpha-Hydroxylase/metabolism , Steroid Hydroxylases/metabolism , Female , Fibroma/enzymology , Granulosa Cell Tumor/enzymology , Humans , Immunohistochemistry , Leydig Cell Tumor/enzymology , Sertoli Cell Tumor/enzymology , Thecoma/enzymologyABSTRACT
In this study, antiserum to lactate dehydrogenase isoenzyme 1 (LD 1) was used to determine immunohistochemical patterns of localization in a variety of germ cell neoplasms of the testis. All 29 seminomas and teratocarcinomas and four of seven embryonal carcinomas were positive for LD 1. These results suggest that elevated serum LD 1 levels noted in patients with germ cell neoplasms may be derived directly from neoplastic tissue and explain the relationship between serum LD 1 levels and tumor burden. A variety of neoplasms from other organs also were evaluated in order to determine the specificity of LD 1 staining for testicular tumors.
Subject(s)
Dysgerminoma/enzymology , L-Lactate Dehydrogenase/metabolism , Testicular Neoplasms/enzymology , Choriocarcinoma/enzymology , Cystadenocarcinoma/enzymology , Female , Granulosa Cell Tumor/enzymology , Histocytochemistry , Humans , Immunochemistry , Immunoenzyme Techniques , Isoenzymes , Leydig Cell Tumor/enzymology , Male , Ovarian Neoplasms/enzymology , Sertoli Cell Tumor/enzymology , Staining and Labeling , Teratoma/enzymologyABSTRACT
In the so-called Sertoli cell syndrome nearly always club shaped invaginations of the basement tubule membrane with intact cellule membrane can be seen. These protuberatons ultrastructurally prove to be multiplicated piles of the lamina densa of the basement membrane. The cells in the Sertoli cell syndrome show a quantitatively and qualitatively reduced pattern of enzymes in contrast with the active Sertoli cells.