Nuclei Of Stromal Cells Research Articles

Image analysis software for automatic DNA ploidy assessment of archival solid tumours

Background: Image cytometry has proved to provide a good alternative to flow cytometry for DNA ploidy measurement of archival tumors. However, when interactively done this technique is unable to give statistically valuable results within an acceptable time for clinical oncology. Methods: An image cytometer was developed for fully automatic DNA ploidy quantitation, focusing efforts on speed and accuracy. Software functionalities include systematic acquisition of fields on a microscopic slide, detection, localization and sorting of nuclei, computation of the DNA content together with post-processing tools, for a deeper analysis of the DNA ploidy diagram. Results: DNA ploidy analysis of archival breast carcinoma samples illustrates the accuracy of DNA ploidy measurements and the sensitivity in the detection of DNA ploidy abnormalities as a result of cell sorting. Conclusions: Fully automatic image cytometry is able to combine qualities of flow cytometry (automatic analysis of a statistically significant collection of cell nuclei) with additional advantages: sorting of unwanted events (debris, stromal and inflammatory cell nuclei) and facilities for an a posteriori control of the quality of cell selection. This method is well suited to DNA ploidy analysis of archival cancer samples. Cytometry 37: 267–274, 1999. © 1999 Wiley-Liss, Inc.

Image analysis software for automatic DNA ploidy assessment of archival solid tumours

Image cytometry has proved to provide a good alternative to flow cytometry for DNA ploidy measurement of archival tumors. However, when interactively done this technique is unable to give statistically valuable results within an acceptable time for clinical oncology. An image cytometer was developed for fully automatic DNA ploidy quantitation, focusing efforts on speed and accuracy. Software functionalities include systematic acquisition of fields on a microscopic slide, detection, localization and sorting of nuclei, computation of the DNA content together with post-processing tools, for a deeper analysis of the DNA ploidy diagram. DNA ploidy analysis of archival breast carcinoma samples illustrates the accuracy of DNA ploidy measurements and the sensitivity in the detection of DNA ploidy abnormalities as a result of cell sorting. Fully automatic image cytometry is able to combine qualities of flow cytometry (automatic analysis of a statistically significant collection of cell nuclei) with additional advantages: sorting of unwanted events (debris, stromal and inflammatory cell nuclei) and facilities for an a posteriori control of the quality of cell selection. This method is well suited to DNA ploidy analysis of archival cancer samples.

Immunolocalization of progesterone receptors in bovine placentomes throughout mid and late gestation and at parturition.

The corpus luteum is the main source of progesterone (P(4)) responsible for maintenance of gestation in cattle. So far it has not been possible to assign any biological role to placental P(4), which contributes only marginally and temporarily to peripheral maternal blood levels. In order to identify possible P(4) target cells within the placenta, placentomes from 150-, 220-, 240-, and 270-day-pregnant cows and from parturient cows (3 animals per group) were screened immunohistochemically for expression of the progesterone receptor (PR). During gestation, PR-positive staining was found exclusively in the nuclei of caruncular stromal cells (CSC; maternal part of the placentome) and of caruncular vascular pericytes. In placentomes from parturient cows, occasional positive nuclear staining was also observed in the walls of small caruncular arteries. The percentage of PR-positive CSC increased slightly from 51.8 +/- 2.6% on Day 150 to 56.2 +/- 5.6% at Day 270 (p < 0.05) and was 58.9 +/- 1.8% at parturition. These results suggest that in pregnant cattle, CSC are under the control of P(4) of placental rather than luteal origin. Thus, whereas luteal P(4) may regulate "coarse" systemic progestational functions in the maternal compartment in the classical hormonal manner, placental P(4) may act as a paracrine factor involved in the local regulation of caruncular growth, differentiation, and functions.

Progestin modulation of c- fos and prolactin gene expression in the human endometrium

Objective: To evaluate the influence of the menstrual cycle and the effects of medroxyprogesterone acetate (MPA) on the expression of the protooncogene c- fos and of prolactin (PRL) in the human endometrium in vivo. Design: Double-blind, placebo-controlled trial. Setting: Healthy volunteers in an academic research environment. Patient(s): Regularly cycling women who were not taking hormonal medication. Intervention(s): Medroxyprogesterone acetate (10 mg/d) or placebo was given for 10 days. Endometrial and blood samples were collected 8–12 hours after the last dose. Main Outcome Measure(s): Immunohistochemical localization of PRL and c- fos in the endometrium, PRL and c- fos messenger RNA levels in the endometrium, and E 2 and progesterone levels in the serum. Result(s): Immunoreactive c- fos was concentrated in the nucleus of stromal cells and was observed in a higher proportion of proliferative endometrial specimens compared with secretory specimens from placebo- or MPA-treated patients. The levels of c- fos messenger RNA were greatly reduced in the secretory endometrium regardless of treatment with placebo or MPA, compared with the proliferative endometrium. The c- fos gene expression correlated positively with the serum E 2 levels ( r = 0.56) and inversely with the progesterone/E 2 ratio ( r = −0.56). The endometrial PRL gene expression (messenger RNA and protein) was rare in the proliferative samples, increased from the early to the mid and late secretory samples, and was increased markedly after treatment with MPA compared with placebo. Conclusion(s): The differentiation of secretory endometrium is accompanied by decreased c- fos and increased PRL gene expression. The inhibition of c- fos gene expression may contribute to the antiproliferative effect of progestins on the endometrium.

Localization of type 5 17beta-hydroxysteroid dehydrogenase, 3beta-hydroxysteroid dehydrogenase, and androgen receptor in the human prostate by in situ hybridization and immunocytochemistry.

An important source of androgens in the human prostate are those synthesized locally from the inactive adrenal precursor dehydroepiandrosterone (DHEA) and its sulfated derivative DHEA-S. Three beta-HSD (hydroxysteroid dehydrogenase) converts DHEA into androstenedione (4-dione), whereas type 5 17beta-HSD catalyzes the reduction of 4-dione into testosterone in the human prostate and other peripheral intracrine tissues. In the present study, we have used two complementary approaches, namely in situ hybridization and immunocytochemistry, to identify the cells that contain the type 5 17beta-HSD messenger RNA and enzyme in human benign prostatic hyperplasia (BPH). Localization of 3beta-HSD and of the androgen receptor (AR) was also investigated by immunostaining in the same tissue. To find out whether there are any differences between BPH and normal prostate tissue, the localization of type 5 17beta-HSD was reexamined by immunocytochemistry in the normal human prostate samples and also in normal prostate epithelial cell line (PrEC). The in situ hybridization results obtained with a tritiated uridine triphosphate (3H-UTP)-labeled type 5 17beta-HSD riboprobe are in agreement with the immunostaining data obtained with a specific antibody to the enzyme. The immunostaining results obtained from normal prostate tissue and BPH were found to be similar. Thus, in the glandular epithelium, basal cells highly express the messenger RNA and the enzyme, whereas luminal cells show a much lower and variable level of expression. In the stroma and walls of blood vessels, fibroblasts and the endothelial cells lining the blood vessels show positive staining. Similar results are observed when the cellular distribution of 3beta-HSD is investigated. AR immunoreactivity, however, shows a different distribution because, in the epithelium, most of the nuclei of basal cells are negative, whereas the majority of nuclei of the luminal cells show positive staining. A strong reaction for AR is also found in most stromal cell nuclei and in the nuclei of most endothelial cells, as well as in some other cells of the walls of blood vessels. In conclusion, human type 5 17beta-HSD, as well as 3beta-HSD, are highly expressed, not only in the basal epithelial cells and stromal fibroblasts but also in the endothelial cells and fibroblasts of the blood vessels. AR, on the other hand, is highly expressed in the luminal cells. The present data suggest that DHEA is transformed in the basal cells of the glandular epithelium into 4-dione by 3beta-HSD and then into testosterone by type 5 17beta-HSD, whereas dihydrotestosterone is synthesized in the luminal cells after diffusion of testosterone from the underlying layer of basal cells. The potential role of androgen formation and action in blood vessels is unknown and opens new avenues of investigation for a better understanding of the multiple roles of androgens.

Sex hormone binding globulin expression and colocalization with estrogen receptor in the human Fallopian tube

The detection of sex hormone binding globulin (SHBG) or SHBG mRNA in several sex steroid target tissues, has raised the possibility that SHBG modulates the action of sex steroids outside the vascular compartment. The presence of SHBG mRNA was investigated by RT-PCR in the poly (A +) RNA fraction of the human Fallopian tube. Human and rat liver were used as positive and negative control tissues, respectively. The electrophoretic analysis of the amplified PCR products showed bands at 219 bp, corresponding to the expected size of the SHBG cDNA, in the Fallopian tube and human liver but not in rat liver, indicating that SHBG might be sinthezised by the Fallopian tube. The cellular localization of SHBG and of estrogen receptor (ER) was examined by immunohystochemistry in consecutive sections of Fallopian tube tissues for individual staining or double immunostaining in the same section. Specific immunostaining of SHBG was present in the epithelial, vascular and muscle cells of the ampullary and isthmic region. In epithelial cells, immunoreactive SHBG was present in the apical end with the highest concentration close to the luminal membrane. The ER was localized in the nuclei of epithelial, stromal and muscle cells of the ampulla and isthmus. Double immunostaining showed that SHBG and ER are colocalized principally in epithelial cells of the ampulla and in muscle cells of the isthmus. In conclusion, the detection of SHBG and SHBG mRNA and the localization of SHBG in estrogen target cells was shown. These findings support the hypothesis that SHBG might regulate sex steroid action at the tissue level.

Expression of the oxytocin receptor in relation to steroid receptors in the uterus of a primate model, the marmoset monkey.

The dynamics of the receptors for oestrogen (ER), progesterone (PR) and oxytocin (OTR) in the marmoset uterus have been analysed throughout the entire cycle and early pregnancy. Uteri obtained during the early, mid/late and late proliferative phase, and the early, mid and late secretory phase and early pregnancy were examined by immunohistochemistry (OTR, ER, PR) and autoradiography (OTR). A massive upregulation of the ER in the cell nuclei of glandular epithelium and stromal cells during the mid proliferative phase was succeeded by a declining staining intensity and positively stained cell number in the secretory phase. PR immunoreactivity increased in the late proliferative phase and early secretory phase, mainly within the cell nuclei, and then declined in both intensity and cell number towards the mid to late secretory phase. Myometrium showed a similar staining pattern for the steroid receptors. OTR were expressed weakly in stroma throughout the entire cycle, increasing slightly in the secretory phase. Glandular epithelium showed positive staining only during the periovulatory period. Myometrial OTR expression was weak during the proliferative phase, increased towards the secretory phase, and was maximal in the late secretory phase. Myometrial tissue adjacent to endometrium was most strongly stained. A cyclic shift evidently occurred in the pattern of steroid receptors, perhaps reflecting the steroid environment or the luteinizing hormone increase associated with ovulation.

Circulating thyroid hormone concentrations and placental thyroid hormone receptor expression in normal human pregnancy and pregnancy complicated by intrauterine growth restriction (IUGR).

Thyroid hormones are critical to growth and development of the human fetus. Abnormal placental development, a major cause of intrauterine growth restriction (IUGR), is associated with a high perinatal mortality and morbidity. Thyroid status has been postulated to play a role in the pathogenesis of such morbidity. In the present study, we have investigated fetal thyroid function and placental expression of thyroid hormone receptor (TR) alpha and beta variants during normal human pregnancy and in pregnancy associated with IUGR. Measurement of free thyroid hormones and TSH concentrations revealed significant rises in free T4 and free T3 between the second and third trimesters of normal pregnancy. Serum concentrations of free T4 and free T3 were lower in fetuses affected by IUGR, although serum TSH levels were not significantly different. Immunocytochemistry demonstrated the presence of TR alpha1, alpha2, and beta1 proteins within the nuclei of trophoblast and stromal placental cells. Immunostaining for these TR variants increased with increasing gestation in normal placenta. Comparison of IUGR placental samples with normal samples revealed greater immunostaining for TR alpha1, alpha2, and beta1 variants in IUGR. Examination of pretranslational expression of TR alpha1, alpha2, beta1, and beta2 variants by semiquantitative RT-PCR revealed increasing expression of TR alpha1, alpha2, and beta2 messenger RNAs with increasing gestation in normal pregnancy, which "mirrored" post-translational expression. However, and in contrast, there were no significant differences in expression of TR messenger RNAs in normal and IUGR placenta. The present findings of reduction in serum free thyroid hormones and increased expression of TR alpha and beta proteins in association with IUGR highlight the potential importance of thyroid status in influencing long-term fetal outcome in this condition.

Stromal cell progesterone and estrogen receptors during proliferation and regression of the decidua basalis in the pregnant rat.

This study examines the distribution and abundance of progesterone receptors (PR) and estrogen receptors (ER) in the decidua basalis (DB) during proliferation (Days 8-12 of gestation) and regression (Days 14-21) in the rat. Stromal cells of the DB and metrial gland exhibited strong nuclear immunostaining for PR throughout gestation. Nuclear localization of ER was detectable only between Days 8-12. The heavily granulated natural killer cells were always negative for PR and ER. DB were dissected between Days 8 and 17 to measure progesterone (P4)-binding sites and receptor proteins by Western blotting. The latter revealed four specific PR isoforms: B (110 kDa), A1 (90 kDa), A2 (76-82 kDa), and C (60-64 kDa). Stromal cell nuclei contained more than 50% of P4-binding sites during DB proliferation but less than 22% during regression (p < 0.05). PR-A and PR-B expression was greatest at proliferative stages (p < 0.05). PR-C increased in relative abundance during DB regression. Two ER isoforms of 66 kDa and 49 kDa were revealed. The 66-kDa ER, the most abundant form, was maximally expressed during proliferation, declining 71% by Day 12 (p < 0.01), whereas the 49-kDa form accounted for up to 90% of ER during regression. Northern blot analysis revealed three prominent transcripts of approximately 11, 7, and 4 kilobases (kb) for PR mRNA, which declined markedly at Days 14 to 17 (p < 0.05), and one of 6.0 kb for ER mRNA, which declined markedly on Day 17 (p < 0.05). Our study establishes that the DB expresses heterogeneity of receptor message and proteins. We propose that preferential expression of receptor isoforms in late pregnancy limits P4 action and promotes DB regression in spite of invariant levels of serum P4, P4-binding sites, and total receptor protein.

Photoperiod-induced changes in androgen receptor expression in testes and accessory sex glands of the bank vole, Clethrionomys glareolus.

Photoperiodic modulation of androgen levels and androgen receptor (AR) expression in testes and accessory sex glands were studied in a seasonally breeding rodent, the bank vole. Juvenile voles subjected to long photoperiod (20L:4D) for 6-8 wk attained sexual maturity, which was associated with a prominent increase in testicular testosterone (T) levels and weight of testes and accessory sex glands. Pubertal development in short photoperiod-treated (6L:18D for 6-8 wk) juveniles was arrested, and subsequently reproductive regression set in with a marked decrease in testicular T levels and gonadal weight. In sexually active voles, strong AR immunostaining was detected in nuclei of epithelial, smooth muscle, and stromal cells of the epididymis, prostate, and seminal vesicles. In active testes, AR was present in nuclei of Sertoli cells, peritubular cells, Leydig cells, and vascular smooth muscle cells. In juveniles, strong to moderate nuclear immunoreactivity was encountered in epithelial and stromal cells of the epididymis and prostate, whereas a weaker reaction was discerned in seminal vesicles. In juvenile testes, AR was localized to vascular smooth muscle cells, peritubular, and interstitial cells. In sexually regressed animals, nuclear staining was almost absent in accessory sex glands, whereas in testes, moderate immunostaining was retained in all other cell types except the Sertoli cells. Western blots of active and regressed testes indicated a marked photoperiod-induced down-regulation of immunodetectable AR in the regressed gonad.

Phyllodes tumor. Fine needle aspiration cytology of eight cases.

To elucidate the fine needle aspiration cytology (FNAC) features of phyllodes tumor (PT). Eight FNAC cases of PT (five benign and three malignant) were reviewed. Features examined were cellularity, epithelial/stromal ratio, phyllodes fragments (PFs), blood vessels crossing PFs, bipolar naked nuclei, stromal pleomorphism, stromal cell size, mitosis, macrophages, sarcomatous component and atypical epithelial component. Benign PTs were characterized by cellular smears with both stromal and epithelial elements. All cases had many highly cellular stromal fragment PFs. Stromal cells were twice the size of a lymphocyte and lacked pleomorphism. Bipolar naked nuclei were a constant feature. The epithelial component predominated, demonstrating benign ductal groups and cellular areas with discohesion superficially mimicking adenocarcinoma in three cases. FNAC of primary malignant PT (one case) had two patterns, one mimicking benign PT and the other demonstrating a pure sarcomatous component. FNAC of metastatic PTs (two cases) showed pure sarcomatous elements. The diagnosis of PT is favored over fibroadenoma when many PFs are present. The size and shape of the stromal cell nuclei were not helpful differentiating features in this study. The diagnosis of malignant PT was based on the purely sarcomatous component.

Expression of parathyroid hormone (PTH)-related peptide (PTHrP) and PTH/PTHrP receptor in giant cell tumour of tendon sheath.

The giant cell tumour of tendon sheath (GCTTS) is mainly composed of mononucleated stromal cells (SC) and multinucleated giant cells (GC), so-called osteoclast-like GC. It is thought that GC are derived from SC, but their precise relationship is not fully understood. Parathyroid hormone (PTH)-related peptide (PTHrP) is now considered to be a cytokine for cell differentiation, which may stimulate osteoclast-like cell formation in haematopoietic cells. Five cases of GCTTS were evaluated immunohistochemically, using a variety of antibodies against PTHrP, PTH/PTHrP receptor, KP-1 as a histiocytic phenotypic antigen, fibronectin as a fibroblastic phenotypic antigen, and proliferating cell nuclear antigen (PCNA). In situ hybridization and immunohistochemistry revealed that in all cases both SC and GC expressed PTHrP. PTH/PTHrP receptor was observed only in histiocytic SC and GC, but not in fibroblastic SC. Almost all GC showed histiocytic features. PCNA immunoreactivity was detected only in the nuclei of SC, and not in GC. Moreover, SC with PTH/PTHrP receptor immunoreactivity were negative for PCNA. These results suggest that GC are derived from histiocytic SC expressing PTH/PTHrP receptor and losing proliferative activity in the process of transition from mononuclear to multinucleated. PTHrP produced by SC and GC may be involved in the formation of osteoclast-like cells in GCTTS by acting in an autocrine/paracrine fashion.

Immunohistochemical profile of phyllodes tumors of the breast

The immunohistochemical profiles of 16 cases of phyllodes tumor of the breast (9 benign and 7 malignant) from 15 patients were examined by the labeled streptavidin biotin method. The expression of Ki-67, p53, bcl-2, alpha-smooth muscle actin (alpha-SMA), desmin, S-100 protein, estrogen receptor (ER), and progesterone receptor (PgR) was analyzed. The number of Ki-67-positive stromal cell nuclei of malignant phyllodes tumor were significantly more prominent than the benign tumors. The p53 expression on the stromal cell nuclei showed a significant difference between malignant and benign cases (86% vs 22%; p<0.05). bcl-2 was regularly seen on the luminal cell cytoplasm and stromal cell labeling showed no significant difference between malignant and benign cases (29% vs 33%). Stromal cells were alpha-SMA positive but refractory among cases, and desmin and S-100 protein were negative. PgR was expressed in all 16 cases and ER in most cases (12/16) the expression of which was restricted to luminal epithelial cell nuclei. These findings indicate that the Ki-67 labeling index and p53 expression in the stroma would be a good diagnostic parameter distinguishing benign tumors from malignant tumors. However, the absence of steroid receptor expression in stromal cells suggests the ineffectiveness of hormonal therapy for phyllodes tumor of the breast.

There are multiple forms of glyceraldehyde-3-phosphate dehydrogenase in prostate cancer cells and normal prostate tissue.

We analyzed glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in normal and malignant human prostate tissues, normal rat prostate, and Dunning R-3327 rat prostate cancer cell lines. We detected multiple forms of GAPDH in Dunning cell lines by two-dimensional protein electrophoresis and Western analysis. Five forms of GAPDH that differed by isoelectric point were detected for each of the two metastatic Dunning cell lines, while four or fewer forms were detected for Dunning cell lines with low metastatic ability. We also detected multiple forms of GAPDH in normal and malignant human prostate specimens by two dimensional protein electrophoresis and immunohistochemical analysis. GAPDH was undetectable in normal human prostate secretory epithelium by immunohistochemistry, but was abundant in nuclei of normal basal cells and stromal cells. In human prostate cancer specimens, there was a rough correlation between cytoplasmic staining for GAPDH and tumor grade, but GAPDH staining was extremely heterogeneous. GAPDH was abundant in nuclei of some high-grade human prostate tumors. Both of the human prostate cancer bone metastases analyzed with immunohistochemistry had markedly elevated cytoplasmic GAPDH expression. We conclude that multiple forms of GAPDH may play diverse roles in normal prostate tissue and in prostate cancer.

There are multiple forms of glyceraldehyde‐3‐phosphate dehydrogenase in prostate cancer cells and normal prostate tissue

We analyzed glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in normal and malignant human prostate tissues, normal rat prostate, and Dunning R-3327 rat prostate cancer cell lines. We detected multiple forms of GAPDH in Dunning cell lines by two-dimensional protein electrophoresis and Western analysis. Five forms of GAPDH that differed by isoelectric point were detected for each of the two metastatic Dunning cell lines, while four or fewer forms were detected for Dunning cell lines with low metastatic ability. We also detected multiple forms of GAPDH in normal and malignant human prostate specimens by two dimensional protein electrophoresis and immunohistochemical analysis. GAPDH was undetectable in normal human prostate secretory epithelium by immunohistochemistry, but was abundant in nuclei of normal basal cells and stromal cells. In human prostate cancer specimens, there was a rough correlation between cytoplasmic staining for GAPDH and tumor grade, but GAPDH staining was extremely heterogeneous. GAPDH was abundant in nuclei of some high-grade human prostate tumors. Both of the human prostate cancer bone metastases analyzed with immunohistochemistry had markedly elevated cytoplasmic GAPDH expression. We conclude that multiple forms of GAPDH may play diverse roles in normal prostate tissue and in prostate cancer. © 1996 Wiley-Liss, Inc.

Immunocytochemical evidence for differential subcellular localization of 5 alpha-reductase isoenzymes in human tissues.

To contrast the previously described distribution of 5 alpha-reductase isoenzyme 2 in human organs we have raised synthetic C-terminal peptides of human 5 alpha-reductase isoenzyme 1 (h 5 alpha r1) and isoenzyme 2 (h 5 alpha r2) respectively, and studied the cellular and subcellular distribution of both isoforms of this key enzyme of testosterone metabolism using the human prostate gland as a reference. h 5 alpha r1, which in Western blots of human prostatic proteins had an apparent molecular weight of 23 kDa, was localized immunohistochemically in the nucleus of prostatic epithelial and stromal cells. Ultrastructurally, it was closely associated with the nuclear matrix. The apparent molecular weight of h 5 alpha r2 was 26 kDa in Western blotting of human prostatic proteins. In immunohistochemically processed sections, it was seen in the cytoplasm of prostatic epithelial as well as stromal cells. An organ screening with genital and extragenital tissues of males and females was performed and to elucidate the distribution of the isoenzymes. Presence of both isoenzymes was demonstrated for a number of tissues, including the central nervous system, the urogenital tract, the respiratory tract, the gastrointestinal tract, skin, and the endocrine system. The divergent localization of 5 alpha-reductase isoenzymes points to potentially different functions in various organs. In view of the nuclear localization of isoenzyme 1, its close spatial relationship with the androgen receptor is presumed to indicate a close association with the receptor mechanism.

Alternative splicing and differential targeting of fibroblast growth factor receptor 1 in the pregnant rat uterus

Recent studies suggest that steroid effects on uterine cell proliferation may be moderated by polypeptide growth factors. We now provide evidence that high affinity fibroblast growth factor (FGF) receptors are present temporally and spatially in the pregnant rat uterus (days 4-6) to support the idea that basic FGF action occurs via binding to its high affinity FGF receptor 1 (FGFR1). Reverse transcription-polymerase chain amplification indicates that both the full-length transcript and an alternatively spliced messenger RNA are present in the uterus. Western immunoblot analysis confirms that rat uterine membrane proteins contain two receptor isoforms, and these receptors bind basic FGF with high affinity and specificity. Immunolocalization of FGFR1 revealed receptor-positive cells in both the uterine stroma and epithelia on days 4-6 of pregnancy. However, the receptor was differentially localized in the disparate cell types. The nuclei of stromal cells were positive for FGFR1, whereas epithelial cell nuclei were negative. Together, these results suggest that FGF signal transduction in uterine stromal cells is mediated by activation of FGFR1.

Alternative splicing and differential targeting of fibroblast growth factor receptor 1 in the pregnant rat uterus.

Recent studies suggest that steroid effects on uterine cell proliferation may be moderated by polypeptide growth factors. We now provide evidence that high affinity fibroblast growth factor (FGF) receptors are present temporally and spatially in the pregnant rat uterus (days 4-6) to support the idea that basic FGF action occurs via binding to its high affinity FGF receptor 1 (FGFR1). Reverse transcription-polymerase chain amplification indicates that both the full-length transcript and an alternatively spliced messenger RNA are present in the uterus. Western immunoblot analysis confirms that rat uterine membrane proteins contain two receptor isoforms, and these receptors bind basic FGF with high affinity and specificity. Immunolocalization of FGFR1 revealed receptor-positive cells in both the uterine stroma and epithelia on days 4-6 of pregnancy. However, the receptor was differentially localized in the disparate cell types. The nuclei of stromal cells were positive for FGFR1, whereas epithelial cell nuclei were negative. Together, these results suggest that FGF signal transduction in uterine stromal cells is mediated by activation of FGFR1.

Pseudoangiomatous Stromal Hyperplasia (PASH)

Pseudoangiomatous stromal hyperplasia (PASH) is frequently a microscopic incidental finding in breast biopsies performed for benign or malignant disease. However, it may also produce a mass lesion. We reviewed PASH seen first as a tumor in 40 women aged 14 to 67 years (mean, 37 years). All but one lesion were clinically palpable. The exceptional tumor was found by mammography. The mass, typically unilateral, was usually diagnosed clinically as a fibroadenoma. Most specimens contained a well-circumscribed tumor with a firm white-gray cut surface. In six cases, there was no discrete gross lesion in the surgical specimen. Microscopically, there was a spectrum of pathological stromal changes ranging from classical PASH with anastomosing slit-shaped spaces outlined by flat, bland spindle cells to more proliferative lesions composed of bundles of plump spindle cells that obscured the underlying pseudoangiomatous architecture in the most florid lesions. The spindle cells were vimentin and CD34 positive and factor VIII negative. In more cellular fascicular lesions, the stromal cells acquired desmin and actin positivity. These immunohistochemical features were consistent with myofibroblastic histogenesis of PASH. Reactivity for progesterone receptor (PR) typically exceeded estrogen receptor (ER) in the nuclei of stromal and glandular cells. In most lesions, the nuclei of stromal spindle cells were ER negative. The majority of the patients were treated by excisional biopsy. One lesion, incompletely excised, spontaneously regressed. One patient had bilateral mastectomies. Follow-up was 0.6-11 years (mean, 4.5 years). Five patients had ipsilateral recurrences, and two had subsequent contralateral PASH. The morphological spectrum of cellular proliferation and staining qualities indicates that the myofibroblast plays a major role in the histogenesis of PASH. The pathogenesis of PASH remains uncertain, but aberrant reactivity of myofibroblasts to endogenous or exogenous hormones is likely to be an important factor. Simple excision is adequate treatment initially and for infrequent recurrences, Diffuse PASH occasionally presents a difficult management problem that may necessitate mastectomy.

Quantitative Analysis of Endometrial Stromal Cells Including Behavioral Changes Related to the Menstrual Cycle in Smear Specimens

We aimed to obtain quantitative data regarding certain characteristics of stromal cells in smear specimens of normal endometrium, including the cytological changes associated with progression through the menstrual cycle. Eleven histological and 25 cytological specimens of normal endometrium were examined immunochemically using monoclonal antibodies to vimentin and keratin by streptavidin-biotin method. As a result, we decided to use the antibody to vimentin for the differential staining of endometrial smears: cells showing positive staining to vimentin were considered stromal cells. The quantitative nuclear measurements in the smear specimens revealed that the nuclei of endometrial stromal cells were more elongated and had a more dispersed distribution than the glandular cells in the proliferative phase. Although a part of the nuclei of stromal cells became rounder in shape in the secretory phase, they were larger in size than those of glandular cells.