Khushbakhat Mittal

Estrogen and progesterone receptors in the uterosacral ligament.

Objective To evaluate steroid hormone receptor status in the uterosacral ligament, a structure that contributes to pelvic support. Methods A descriptive study was conducted by sampling the uterosacral ligaments from 25 consecutive women undergoing hysterectomy by the primary author for nonmalignant conditions. Using immunohistochemical staining techniques, uterosacral ligaments were assessed for the presence and location of estrogen and progesterone receptors. Positive and negative controls were used. Confirmation of the uterosacral ligament was performed histologically. Results Using commercially available monoclonal antibodies, estrogen and progesterone receptors were detected in the nuclei of smooth muscle cells of the uterosacral ligament in all patients, regardless of variations in age, race, menopausal status, parity, body mass index, and medications affecting serum steroid hormone levels. Hormone receptors were not found in the collagen, vascular, or neuronal components. Conclusion The presence of estrogen and progesterone receptors in the uterosacral ligaments means that this structure may be a target for estrogen and progesterone. This finding might suggest a possible role for steroid hormones in pelvic support.

Cesarean scar pregnancy and early placenta accreta share common histology

To determine, by evaluation of histological slides, images and descriptions of early (second‐trimester) placenta accreta (EPA) and placental implantation in cases of Cesarean scar pregnancy (CSP), whether these are pathologically indistinguishable and whether they both represent different stages in the disease continuum leading to morbidly adherent placenta in the third trimester.

Utility of p16INK4a, CEA, Ki67, P53 and ER/PR in the differential diagnosis of benign, premalignant, and malignant glandular lesions of the uterine cervix and their relationship with Silverberg scoring system for endocervical glandular lesions.

Early detection of premalignant and malignant glandular lesions of the uterine cervix and their distinction from benign mimics is crucial but sometimes difficult. In this study, we investigated utility of expression of p16INK4a, CEA, Ki67, p53 and ER/PR in evaluating the benign, premalignant, and malignant glandular lesions of the uterine cervix. A total of 35 cervical cone or LEEP cases were collected including 14 adenocarcinoma in situ (AIS), 7 endocervical glandular dysplasia (EGD), and 14 benign mimics (BM). A histological score for each case according to the criteria proposed by Silverberg group was assigned independently by 4 pathologists. Formalin-fixed, paraffin-embedded sections were immunostained with p16, CEA, p53, Ki67, and ER/PR. Immunoreactivity was scored based on intensity (0 = none, 1 = mild, 2 = moderate, 3 = marked) and percentage of cells staining (0 = <1%,1 = 2-10%, 2 = 11-40%, 3 = >40%). A comparison of histological scores and immunoscores in 3 diagnostic categories was analysed. The histological scores assigned independently by 4 pathologists were all equal or above 6 for AIS, between 3 and 5 for EGD, and equal or below 3 for BM. There was increased expression of p16 and CEA in EGD compared with BM (P < 0.05), with further increase in expression of these markers in AIS compared with EGD (P < 0.05). Ki67 expression was significantly increased in AIS compared to EGD (P < 0.05) as well as compared to BM (P < 0.05). Ki67 expression was only slightly increased in EGD as compared to BM. There was a loss of ER/PR in cervical AIS, but not in EGD. Our results indicate that the Silverberg scoring system is a useful tool in differential diagnosis of cervical glandular lesions for increased diagnostic accuracy and interobserver agreement. Most cervical glandular lesions can be differentiated by using a combination of histological scores with a panel of immunomarkers.

Inhibitors of SCF-Skp2/Cks1 E3 Ligase Block Estrogen-Induced Growth Stimulation and Degradation of Nuclear p27kip1: Therapeutic Potential for Endometrial Cancer

In many human cancers, the tumor suppressor, p27(kip1) (p27), a cyclin-dependent kinase inhibitor critical to cell cycle arrest, undergoes perpetual ubiquitin-mediated proteasomal degradation by the E3 ligase complex SCF-Skp2/Cks1 and/or cytoplasmic mislocalization. Lack of nuclear p27 causes aberrant cell cycle progression, and cytoplasmic p27 mediates cell migration/metastasis. We previously showed that mitogenic 17-β-estradiol (E2) induces degradation of p27 by the E3 ligase Skp1-Cullin1-F-Box- S phase kinase-associated protein2/cyclin dependent kinase regulatory subunit 1 in primary endometrial epithelial cells and endometrial carcinoma (ECA) cell lines, suggesting a pathogenic mechanism for type I ECA, an E2-induced cancer. The current studies show that treatment of endometrial carcinoma cells-1 (ECC-1) with small molecule inhibitors of Skp2/Cks1 E3 ligase activity (Skp2E3LIs) stabilizes p27 in the nucleus, decreases p27 in the cytoplasm, and prevents E2-induced proliferation and degradation of p27 in endometrial carcinoma cells-1 and primary ECA cells. Furthermore, Skp2E3LIs increase p27 half-life by 6 hours, inhibit cell proliferation (IC50, 14.3μM), block retinoblastoma protein (pRB) phosphorylation, induce G1 phase block, and are not cytotoxic. Similarly, using super resolution fluorescence localization microscopy and quantification, Skp2E3LIs increase p27 protein in the nucleus by 1.8-fold. In vivo, injection of Skp2E3LIs significantly increases nuclear p27 and reduces proliferation of endometrial epithelial cells by 42%-62% in ovariectomized E2-primed mice. Skp2E3LIs are specific inhibitors of proteolytic degradation that pharmacologically target the binding interaction between the E3 ligase, SCF-Skp2/Cks1, and p27 to stabilize nuclear p27 and prevent cell cycle progression. These targeted inhibitors have the potential to be an important therapeutic advance over general proteasome inhibitors for cancers characterized by SCF-Skp2/Cks1-mediated destruction of nuclear p27.

Lymphomatoid granulomatosis in the acquired immunodeficiency syndrome. Evidence of Epstein-Barr virus infection and B-cell clonal selection without myc rearrangement

A case of lymphomatoid granulomatosis of lung that occurred in a patient with the acquired immunodeficiency syndrome (AIDS) was studied by light microscope, electron microscope, cell surface markers, and Southern blot test. Clonal selection of two clones of B‐cells was seen. Two clones were infected with Epstein‐Barr virus. There was no c‐myc rearrangement. Lymphomatoid granulomatosis in patients with AIDS may represent multiclonal selection of B‐lymphocytes in association with Epstein‐Barr virus infection.

Estrogen and progesterone regulate p27kip1 levels via the ubiquitin-proteasome system: pathogenic and therapeutic implications for endometrial cancer.

The levels of proteins that control the cell cycle are regulated by ubiquitin-mediated degradation via the ubiquitin-proteasome system (UPS) by substrate-specific E3 ubiquitin ligases. The cyclin-dependent kinase inhibitor, p27kip1 (p27), that blocks the cell cycle in G1, is ubiquitylated by the E3 ligase SCF-Skp2/Cks1 for degradation by the UPS. In turn, Skp2 and Cks1 are ubiquitylated by the E3 ligase complex APC/Cdh1 for destruction thereby maintaining abundant levels of nuclear p27. We previously showed that perpetual proteasomal degradation of p27 is an early event in Type I endometrial carcinogenesis (ECA), an estrogen (E2)-induced cancer. The present studies demonstrate that E2 stimulates growth of ECA cell lines and normal primary endometrial epithelial cells (EECs) and induces MAPK-ERK1/2-dependent phosphorylation of p27 on Thr187, a prerequisite for p27 ubiquitylation by nuclear SCF-Skp2/Cks1 and subsequent degradation. In addition, E2 decreases the E3 ligase [APC]Cdh1 leaving Skp2 and Cks1 intact to cause p27 degradation. Furthermore, knocking-down Skp2 prevents E2-induced p27 degradation and growth stimulation suggesting that the pathogenesis of E2-induced ECA is dependent on Skp2-mediated degradation of p27. Conversely, progesterone (Pg) as an inhibitor of endometrial proliferation increases nuclear p27 and Cdh1 in primary EECs and ECA cells. Pg, also increases Cdh1 binding to APC to form the active E3ligase. Knocking-down Cdh1 obviates Pg-induced stabilization of p27 and growth inhibition. Notably, neither E2 nor Pg affected transcription of Cdh1, Skp2, Cks1 nor p27. These studies provide new insights into hormone regulation of cell proliferation through the UPS. The data implicates that preventing nuclear p27 degradation by blocking Skp2/Cks1-mediated degradation of p27 or increasing Cdh1 to mediate degradation of Skp2-Cks1 are potential strategies for the prevention and treatment of ECA.

Squamous Cell Carcinoma in HIV‐Positive Patients Under Age 45

Eight patients 45 years of age and under (range, 29 to 45) with squamous cell carcinoma of the head and neck and infection with the human immunodeficiency virus are reported. Primary tumor sites include nasopharynx, oral cavity, oropharynx, and larynx. Probes for the human papillomavirus were positive in two of the patients. Therapy consisted of surgery followed by radiation therapy in five patients, surgery alone in one patient, and radiation therapy in the remaining two patients. Follow‐up ranged up to 2 years and revealed four deaths, three patients alive without disease, and one patient recently posttreatment with residual disease. The significance of the presence of the human papillomavirus in these individuals remains to be determined. The incidence of squamous cell carcinoma of the head and neck under age 45 is low, and whether there may be a higher incidence in HIV‐positive patients cannot be determined from this small series.

Patterns of mRNA for epidermal growth factor receptor and keratin B-2 in normal cervical epithelium and in cervical intraepithelial neoplasia

Patterns of epidermal growth factor receptor (EGFR) and keratin B-2 (KB2) mRNA localization were studied in samples of normal cervical squamous epithelium and in samples of cervical intraepithelial neoplasia (CIN). 35S-Labeled EGFR and KB2 DNA probes were used for in situ hybridization on formalin-fixed tissue sections. Normal cervical squamous epithelium showed predominantly basal and parabasal expression of EGFR mRNA and suprabasal and midepithelial localization of KB2 mRNA. CIN lesions with moderate to severe dysplasia were generally characterized by continued expression of EGFR mRNA and decreased KB2 mRNA in midepithelial locations. Ratios of KB2 mRNA levels at the basal layer to KB2 mRNA levels at the midepithelial location were increased in moderate and severe dysplasia (CIN II and III) compared with normal epithelium (P less than .01). Ratios of EGFR mRNA levels at the midepithelial level to those of EGFR mRNA at the basal layer were increased in moderate to severe dysplasia compared with normal epithelium. These findings indicate a possible in vivo role of EGFR gene expression in normal and neoplastic proliferation and in prevention of differentiation of the cervical epithelium.

Lymphoma of ovary with stromal luteinization, presenting as secondary amenorrhea

A case of primary lymphoma of the ovary with extensive stromal luteinization is presented. Immunohistochemical markers that work satisfactorily in fixed tissues allowed a diagnosis of B cell lymphoma to be made on formalin-fixed tissue sections. Immunohistochemical analysis of frozen tissue and genotypic analysis confirmed the diagnosis of B cell lymphoma. By electron microscopy two populations of cells, including large atypical lymphoid cells and luteinized stromal cells, were identified. Presentation with amenorrhea and resumption of menstrual periods after removal of the tumor suggested the possibility of functional activity in the luteinized stromal cells. Stromal luteinization has not been previously described in primary ovarian lymphoma.

Flow cytometric DNA ploidy and quantitative histopathology in partial moles.

SummaryThis study quantitates morphologic changes seen in partial moles and hydropic abortuses in an attempt to find a correlation with DNA content. Thirty-two products of conception were studied. Fifteen were diagnosed as definitive partial moles (DPM), and 17 had changes suggestive of a partial mole (SPM). We determined DNA ploidy by flow cytometry and quantitatively analyzed the following microscopic features: villous edema and sclerosis, central cistern formation, villous blood vessels, trophoblastic proliferation, and trophoblastic inclusions. No single pathologic feature significantly correlated with DNA content, even though triploid cases showed more inclusions than diploid cases (8.3 versus 7.2 on the average per ten X100 fields). An overall diagnosis of DPM did correlate with ploidy; 12 of 18 triploid cases (67%) compared with only three of 13 diploid cases (23%) had been previously diagnosed as DPM (p < 0.05). Flow cytometric DNA ploidy estimates DNA content and cannot determine chromosomal origin. It is possible that detailed cytogenetic studies of moles compared with abortuses may show a significant correlation of the proportion of paternal chromosomes, morphologic changes, and the risk of persistent gestational trophoblastic disease.