Sep 2017 DOI 10.14302/issn.2572-3030.jcgb-17-1737
J. O OgunbiyiCorresponding author
Scientific review, Supervisory, slides evaluation and final review
Objective P16INK4a and Ki-67 are adjuncts to current histological assessment of cervical biopsies in identifying cases that require strict follow up and prompt intervention. This study aimed to evaluate P16INK4a and Ki-67 expression in squamous intraepithelial and other benign cervical lesions. Methods A retrospective cross-sectional study of 153 cases of cervical biopsies diagnosed as CIN and benign cervical lesions between 2006 and 2013 at the University College Hospital, Ibadan, Nigeria. Slides and tissue blocks of all the selected cases were retrieved and classified using the 2003 WHO classification for intraepithelial and benign cervical lesions and were stained with p16INK4a and Ki-67 immunohistochemical stains following heat-induced antigen retrieval. Results were evaluated and compared with histologic diagnosis. Results Cases were classified as chronic cervicitis (12.3%), squamous Metaplasia (0.7%), CIN 1 (47.1%), CIN 2 (36.6%) and CIN 3 (3.3%). Majority of the non-dysplastic cervical lesions (including chronic cervicitis cases) showed low P16INK4a reactivity. Positive P16INK4a reactivity was seen in 80% of CIN 3 cases, 83.9% of CIN 2 cases, and, surprisingly, in 97.2% of CIN 1 cases. Ki-67 positivity was seen in 36.6% of cases (75% CIN 2 and 60% CIN 3). There was a significant correlation between the H&E diagnoses of CIN and P16INK4a/Ki-67 immunoreactivities. Conclusion Majority of the CIN 1 cases showing low grade p16INK4a immunereactivity strongly suggesting that cervical squamous intraepithelial neoplasia in this environment is likely associated with high grade HPV subtype infections and may predict possible progression to high grade squamous intraepithelial neoplasia. The use of P16INK4a and Ki-67 in the evaluation of cervical biopsies for benign mimics of high grade intraepithelial lesion will aid proper single Pathologist evaluation and help in patients triaging for follow up.
Oct 2015
S. Woo JenniferCorresponding author
Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, 10833 Le Conte Avenue, CHS 13-145, Los Angeles, CA, United States
The mitotic count is the most frequent reason for discordance between pathologists in modified Bloom and Richardson (mBR) scoring. Recently, the phosphohistone H3 (PHH3) immunohistochemical stain has been proposed as a potential surrogate marker for mitotic figures. This study examines the differences between H&E mitotic count, PHH3 mitotic count, and Ki-67 index in invasive breast carcinoma. A retrospective review of invasive breast carcinoma cases from 2013- 2014 was performed. H&E and PHH3 mitotic counts were assigned a mitotic score of 1 to 3 using mBR criteria. Ki-67 index was categorized into a three-grade system: <10% (low), 10 - <20% (intermediate), and >20% (high). A total of 451 cases were evaluated. PHH3 versus H&E mitotic count changed mBR scores in 24% of cases, upgrading in 23% and downgrading in 1%. A total of 431 cases had both Ki-67 and PHH3 available for comparison. Both H&E and PHH3 mitotic scores correlated with Ki-67 in 51% of cases; however, PHH3 had better correlation. We conclude that PHH3 in breast carcinoma allows for a more sensitive and practical approach in the identification of mitotic figures. PHH3 IHC is useful as a confirmatory tool in assessing the final mitotic score for more accurate mBR scoring and grading. In this study, 48 out of 451 (10.6%) of patients had a significant upgrade that may change the patient's treatment plans, including the addition of chemotherapy
Jan 2022 DOI 10.14302/issn.2372-6601.jhor-22-4061
B. Danilova AnnaCorresponding author
N.N. Petrov National Medicine Research Center of Oncology, Department of Oncoimmunology, 197758, Leningradskaya str., 68, Pesochny, Saint-Petersburg, Russian Federation
Background Human malignant cell models which reflect the structural and physiological complexity of tumor tissue are of great importance for preclinical research in oncology. Spheroids/tumoroids derived from solid tumors are of great interest as cellular models mimicking the first vascular-free growth phase of a tumor node. The fact of the identity between artificially created tumor multicellular aggregates and the real tumor tissue, however, needs to be specified, described and validated in order to see how closely the spheroids are biologically similar to the malignized tissues in vivo compared to the monolayer cell cultures traditionally used. We present here a comparison study of the characteristics of solid tumor cells of different histogenesis (melanomas, soft tissue sarcomas and bone sarcomas, epithelial tumors) cultured in two dimensions (monolayer culture) and three dimensional space (spheroid), namely: spatial organization, multiplication, metabolic activity. Patients and Methods For the creation of 2 D and 3D cell models the cells isolated from the patient's solid tumor fragments obtained intraoperatively were used. 15 samples of skin melanoma, 20 samples of soft tissue and osteogenic sarcomas (STBS), and 9 samples of epithelial tumors (ET). The tumor cells were all cultivated for at least 10 passages. We used phase contrast, confocal microscopy, and immunohistochemistry to investigate spheroids and monolayer cultures. The supernatants of tumor cells grown in 2D and 3D cultures were studied using ELISA and multiplex analysis for the production of a spectrum of chemokines and cytokines supporting the immunosuppression, invasion and metastasis processes. Results Tumor specimens received were predominantly of metastatic origin (75%). In 100% of cases 2D cultures were received, in 88.6% of cases (39 out of 44) we succeeded in obtaining spheroids. There was no direct correlation between the efficiency of tumoroid formation and the tumor's histogenetic origin and the stage of the cancer process (primary tumor, recurrence, metastasis). The median size of spheroids by 4-5 days of cultivation with a starting concentration of 10000 cells per well was 657.14 μm for melanoma (min 400 - max 1000 μm), 571.42 μm (min 400 - max 700 μm), 507.14 μm (min 300 - max 600 μm) for soft tissue sarcomas, 650.0 μm (min 400 - max 900 μm) for osteogenic sarcomas. Immunochemical analysis of Ki-67, GLUT1, and Ecadherin markers was carried out for tumor tissue samples, single-layer tumor cultures, and tumoroids of every patient. The distribution of the stained groups in the spheroids was distinct from the monolayer cultures and more in accordance with the distribution of such in the tissue tumor, the number of Ki-67+ cells was increasing in the spheroids. We detected no dependence of Ki-67+ and GLUT1+ cell localization grade on spheroid size. We identified E-cadherin in tumor tissue and tumoroids of breast carcinoma and one melanoma culture. Monolayer cultures did not express it. The increase in secretory cell activity of the solid tumor cells from 2D to 3D system was observed when CCL2, CCL3, CXCL1, CXCL16, MIF, IL10, MICA (p<0.01) were investigated. Conclusion The presence of patient-specific cells of solid tumors in a 3D environment causes activation of the proliferative and metabolic processes as compared to monolayer cultures, which makes these models approximate the real world clinical picture. The production of chemokines that can attract to the tumor various types of immune system cells, to include their immature versions, as well as production of cytokines and Immunosuppression factors that, when present in the tumor microenvironment in the high concentrations, contribute to the formation of immune cells having suppressive capacities occurs in the 3D cell system. Three-dimensional model of the initial tumor nodule formation stage thus demonstrates the forming process of tumor cells favorable for them microenvironment. Construction of three-dimensional models - spheroids of tumor cells of differing histogenesis demands individual approach and more thorough investigation.