Expression of stem cell biomarker CD44 in oral squamous cell carcinoma and its association with lymph node metastasis and TNM staging.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is one of the most prevalent cancers in the world. OSCC is a highly invasive lesion frequently having soaring morbidity as well as substantial mortality, attributed to resistance to therapy, metastasis, and recurrence driven by specific populations of cancer stem cells (CSC). The evidence of the association of expression of stem cell biomarker CD44 and metastatic potential of the tumor is inconclusive in OSCC and hence needs further evaluation. OBJECTIVES: To determine the immunohistochemical expression of CD44 in OSCC and to find its association with lymph node metastasis and TNM staging. MATERIALS AND METHODS: One hundred and five histologically proven cases of OSCC were studied. Histopathological parameters like depth of invasion, presence of lymph node metastasis, grading, and TNM staging were done according to the new AJCC staging criteria. Both intensity and proportion of CD44 expression were recorded. RESULTS: The mean age observed in this study was 52.59 years with a male: female ratio of 1:3.76. Forty-nine cases (46.6%) showed a depth of invasion of more than 10 mm. Fifty-two out of one hundred and five cases (49%) had nodal involvement. TNM staging was 5.7%, 7.6%, 44.7%, and 42% for stages I, II, III, and IV, respectively. The majority of the cases (87.5%) showed CD44 expression in the tumor. There was a significant association between the CD44 expression and lymph node metastases (P < 0.001). Higher CD44 expression was seen in stages III and IV (P < 0.001). CONCLUSION: CD44, a stem cell biomarker is significantly associated with higher TNM stage and lymph node metastases. This may be useful in predicting the tumor behavior in the small biopsy.

Association Between the Immunohistochemistry Expression of E-cadherin, Beta-Catenin, and CD44 in Colorectal Adenocarcinoma.

Background Colorectal cancer is a leading cause of cancer-related deaths worldwide, and epithelial-mesenchymal transition (EMT) plays an important role in cancer metastasis. In EMT, there is downregulation of E-cadherin, an intracellular adhesion molecule, as well as mutations in beta-catenin genes. On immunohistochemistry (IHC), the expression of CD44 portrays stem cell differentiation, which, in turn, is strongly associated with EMT. Thus, newer targeted therapies can be advised based on the expression of EMT and stem cell differentiation. Aims and objectives To determine the IHC expression of E-cadherin, beta-catenin, and CD44 in colorectal adenocarcinoma and find the association of the IHC expression of E-cadherin, beta-catenin, and CD44 with the histopathological grade, stage, lymph node metastasis, and lymphovascular invasion of colorectal adenocarcinoma. Materials and methods Fifty histologically proven cases of colorectal adenocarcinoma from 2016 to 2021 were included in this study, and clinicopathological data including age, gender, grading, TNM (tumour, node, and metastasis) staging, and lymph node metastasis were collected and hematoxylin and eosin slides were reviewed. IHC staining for E-cadherin, beta-catenin, and CD44 was done for all cases using the peroxidase and anti-peroxidase method, and the results were analysed. Results Peak incidence occurred in the 61-70 years age group (36%), and the most common site of the tumour was the rectal area (48%). The majority of the cases were in TNM stage II (37.3%), and a low expression of E-cadherin was found to be associated with higher T stage (p = 0.03), TNM staging (p = 0.04), as well as the presence of lymph node metastasis (p = 0.006). High beta-catenin expression was observed to have a significant correlation with a higher T stage (p = 0.006) and TNM staging (p = 0.005), while high CD44 expression was found to be associated with lymph node metastasis (p = 0.01). Altered expression of EMT-related proteins (E-cadherin and beta-catenin) showed a significant correlation with higher T stage (p = 0.03), TNM staging (p = 0.016), and lymph node metastasis (0.04). Conclusions EMT and cancer stem cell IHC markers are biomarkers for aggressive tumour growth and lymph node metastasis. Hence, EMT markers (E-cadherin and beta-catenin) and cancer stem cell markers (CD44) can be used as prognostic markers.

Outcome of Treatment in Verrucous Carcinoma of Oral Cavity: A Tertiary Rural Hospital Experience.

Verrucous carcinoma of oral cavity is a highly well differentiated variant of squamous cell carcinoma with a low potential for invasion and metastasis. It is prevalent in the tobacco quid chewing population in our region. In this observational study, we reviewed the medical case records of 58 patients treated for oral verrucous carcinoma staged T2 to T4a. All patients underwent wide excision of tumour which included marginal mandibulectomy in 22 and hemimandibulectomy in 23 patients along with neck dissection saving the accessory nerve and internal jugular vein. 5 patients were found to have bone involvement along the alveolar sockets. 11 patients had other associated premalignant lesions in oral cavity. Only 2 patients had lymph node metastasis without extra nodal spread in submandibular region. With a mean follow up of 6 years and minimum follow up of 1 year, 3 patients had local recurrence. All these 3 patients had bone involvement and 2 of them had lymph node metastasis on histopathological examination. 3 patients who had associated premalignant lesions developed second primary in oral cavity after 3 years. In our experience, verrucous carcinoma has good prognosis when treated by surgery. Bone involvement along alveolar sockets and associated oral premalignant lesions adversely affect the outcome. There was no difference in the outcome between selective and modified radical neck dissection. Therefore selective neck dissection (supraomohyoid) would be adequate in treating these patients. Adjuvant radiotherapy can be reserved for T4a lesions or for positive margins.

Evaluation of Tumour Volume as a Prognostic Factor in Carcinoma Buccal Mucosa.

(1) To determine tumor volume by clinical measurement, imaging and histopathological examination in patients with squamous carcinoma of buccal mucosa. (2) To correlate tumor volume with lymph-node metastasis and loco-regional control. Retrospective Observational Study. Rural tertiary care hospital. 75 Patients undergoing surgery for T2 (68%) and T3 (32%) Oral squamous carcinomas were included in this observational study. Tumor volume-a product of maximum length, breadth and thickness, was determined clinically and by imaging. Tumor volume on histopathology was documented and correlated with lymph-node metastasis. After 13 months minimum follow-up, tumor volume was correlated with loco-regional control. Recurrences were analysed with regard to tumor volume, depth of invasion, lymph-node metastasis and resection margins. Average tumor volume was 7.1 cm3among T2 and 14.4 cm3among T3. 25% of T2 tumors had metastatic lymph-nodes compared to 33.3% in T3. There was positive correlation between tumor volume and lymph-node metastasis. Lymph-node metastasis was absent when tumor volume was < 8 cm3. There were 4 local and 2 regional recurrences in this study. Average histopathological tumor volume among patients who had local recurrence was 12.95 cm3. Regional recurrences occurred in tumors staged N2b and N3b on histopathology. Average depth of invasion in patients with recurrence was 10.33 mm.. Tumor volume represents actual tumor load and correlates with lymph node metastasis and prognosis in oral cancers. Depth of invasion and tumor thickness influence staging and prognosis. Large volume tumors have poor oncological outcome.

Expression of Vascular Endothelial Growth Factor and Microvessel Density in Psoriatic Skin Lesions.

BACKGROUND: Psoriasis is a common chronic and immune-mediated skin disorder having a significant impact on the patient's quality of life. An analysis of the role of angiogenic vascular endothelial growth factor (VEGF) expression and microvessel density in psoriatic skin lesions may help in better understanding of the disease pathogenesis. The aim of this study was to examine the expression of vascular endothelial growth factor (VEGF) and microvessel density using CD34 antibodies in psoriatic skin lesions by immunohistochemical examination using normal skin of healthy individuals as controls. MATERIALS AND METHODS: Patients with clinical diagnosis of plaque type of psoriasis (e.g., chronic plaque) (n = 49) were included in the study. 5-mm punch biopsies were taken from the psoriatic skin lesions in these patients. A total of 20 punch biopsies were taken from the control group comprising of 20 healthy volunteers. The biopsies were subjected to histopathological examination for confirmation of diagnosis and grading. Immunohistochemical evaluation was done for the expression of VEGF, and microvessel density was assessed using CD34 and compared with the controls. RESULTS: An increased VEGF expression by keratinocytes (49.80% ± 21.16%) and microvessel density in the papillary dermis (15.302% ± 3.8061%) was observed in patients with psoriasis, which was significantly higher as compared to controls (P < 0.0001). A significant positive correlation was observed between VEGF expression by keratinocytes and the microvessel density in the dermis (r = 0.664, P = 0.01). No significant correlation was observedbetween the histopathological grade of psoriasis and microvessel density, or with the VEGF expression. CONCLUSION: VEGF expression ascertained to be a significant factor in the pathogenesis of psoriasis.

Outcome of compartment resection of locally advanced oral cancers extending to infratemporal fossa: a tertiary rural hospital experience.

Locally advanced oral cancers extending to infratemporal fossa (ITF) are a challenge to head and neck surgeons. These tumors are classified as T4b whenever the masticator space (MS), pterygoid muscles (PM), and pterygoid plates (PP) are involved according to AJCC classification. Until recently, these tumors were considered inoperable and treated only with palliative intent. However, a few studies in the last decade showed that many of these tumors could be resected with a reasonably favorable prognosis by compartment resection of ITF, particularly when the tumor was below sigmoid notch of mandible. A few studies attempted to downstage these tumors by neo-adjuvant chemotherapy before attempting resection. Oral Squamous cell carcinoma has a high prevalence in South India. Majority of these patients are females addicted to tobacco quid chewing and present with locally advanced disease. In this retrospective analysis, we evaluated the outcome of treatment of oral squamous cell carcinoma extending to ITF and staged T4b in 52 patients. All patients underwent Composite resection including compartment resection of ITF followed by adjuvant treatment. 20 patients had received neo-adjuvant chemotherapy. Pectoralis major myocutaneous flap was the mainstay of reconstruction. After mean follow-up of 2 years, 31 patients are alive and disease free. 14 patients had local recurrence in ITF and 2 patients had recurred in cervical nodes. 8 patients died due to disease and 6 are on palliative care. Neo-adjuvant chemotherapy did not benefit the outcome. Close margins of resection, extra nodal spread from lymph nodes and supra notch and involvement of posterior part of ITF were factors which predisposed to recurrence.

GSTP1 c.341C>T gene polymorphism increases the risk of oral squamous cell carcinoma.

Glutathione S Transferases (GST) are anti-oxidant enzymes involved in detoxification of cellular and exogenous carcinogens and oxidative products of reactive oxygen species. Genetic polymorphisms can attenuate the detoxification capacity of GST and consequently increase the susceptibility to carcinogenesis. There are eight classes of GST enzymes of which pi subtype is the predominant form expressed in the oral mucosa. c.341C > T single nucleotide polymorphism (rs1138272) in GSTP1 gene, is a functional variation that reduces the enzymatic activity of GST pi. We carried out a 1:2 case-control study involving 270 individuals to determine the association of c.341C > T variation with the risk of oral squamous cell carcinoma. GSTP1 c.341C > T variation was genotyped by PCR-RFLP method. GST pi expression in the tumour sample was determined by immunohistochemistry. Tobacco consumption was the major risk factor among cancer patients. The odds ratio for the risk of oral squamous cell carcinoma in individuals with the minor allele was 4.5 (0.95 CI = 2.3-8.9; P = 0.000004). The genotype was found to follow dominant mode of inheritance (OR 4.4 [0.95 CI = 2.1-9.2]; P = 0.00006). Our results support the conclusion that c.341C > T variation in GSTP1 increases the risk of OSCC in patients habituated to tobacco consumption.

Cytological Evaluation of Thyroid Lesions by Nuclear Morphology and Nuclear Morphometry.

INTRODUCTION: Fine needle aspiration (FNA) of the thyroid gland is an effective diagnostic method. The Bethesda system for reporting thyroid cytopathology classifies them into six categories and gives implied risk for malignancy and management protocol in each category. Though the system gives specific criteria, diagnostic dilemma still exists. Using nuclear morphometry, we can quantify the number of parameters, such as those related to nuclear size and shape. The evaluation of nuclear morphometry is not well established in thyroid cytology. OBJECTIVE: To classify thyroid lesions on fine needle aspiration cytology (FNAC) using Bethesda system and to evaluate the significance of nuclear parameters in improving the prediction of thyroid malignancy. MATERIALS AND METHODS: In the present study, 120 FNAC cases of thyroid lesions with histological diagnosis were included. Computerized nuclear morphometry was done on 81 cases which had confirmed cytohistological correlation, using Aperio computer software. One hundred nuclei from each case were outlined and eight nuclear parameters were analyzed. RESULTS: In the present study, thyroid lesions were common in female with M: F ratio of 1:5 and most commonly in 40-60 yrs. Under Bethesda system, 73 (60.83%) were category II; 14 (11.6%) were category III, 3 (2.5%) were category IV, 8 (6.6%) were category V, and 22 (18.3%) were category VI, which were malignant on histopathological correlation. Sensitivity, specificity, and diagnostic accuracy of Bethesda reporting system are 62.5, 84.38, and 74.16%, respectively. Minimal nuclear diameter, maximal nuclear diameter, nuclear perimeter, and nuclear area were higher in malignant group compared to nonneoplastic and benign group. CONCLUSION: The Bethesda system is a useful standardized system of reporting thyroid cytopathology. It gives implied risk of malignancy. Nuclear morphometry by computerized image analysis can be utilized as an additional diagnostic tool.

Clinicopathological study of surgical margins in squamous cell carcinoma of buccal mucosa.

OBJECTIVE: To evaluate the margins of resected specimen of oral squamous cell carcinoma (SCC) and to document the surgical margin (measured at the time of resection) and margins at the time of pathological examination (after immersion of the specimen in formalin). METHODS: Patients who were diagnosed and confirmed with squamous carcinoma of buccal mucosa were included in the study. Patients underwent resection of the tumor with a margin of 1 cm. Soon after resection, the distance between outermost visible margin of the tumor and the margin of the specimen was measured and documented. Specimens were fixed in 10% formalin and submitted for gross and histopathological examination. The closest histopathologic margin was compared with the in situ margin (10 mm) to determine and document any shrinkage of the margin and the percentage of discrepancy if any. RESULTS: A total of 52 specimens were collected from patients between January 2014 and December 2014. All specimens were obtained from the oral cavity (n = 52) of which 43 (82.7%) were squamous cell carcinoma and 9 (17.3%) were verrucous variant of squamous cell carcinoma. The average decrease in tumor margins measured after fixation in formalin was found to be statistically significant (P < 0.05) in 65% of cases. CONCLUSION: Tumor margin shrinks significantly after formalin fixation by about 25%. The operating surgeon and pathologist should be well aware of such changes while planning for further management thereby ensuring adequate margin of resection and adjuvant treatment wherever required to prevent possible local recurrence of the disease.

Evaluation of histomorphological and immunohistochemical parameters as biomarkers of cervical lymph node metastasis in squamous cell carcinoma of oral cavity: A retrospective study.

INTRODUCTION: Oral squamous cell carcinoma (OSCC) is seen worldwide but is more common in India. Lymph node (LN) metastasis has been shown to be the strongest prognostic factor in OSCC. Many histopathological and immunohistochemical markers have been studied to predict the LN metastasis. AIMS: To identify clinicopathological factors and immunohistochemical (IHC) biomarkers which predict cervical metastasis in OSCC patients. STUDY AND DESIGN: A total of 105 cases of OSCC were taken up for our study. Histopathological parameters such as tumor thickness, depth, degree of differentiation, pattern of invasion (POI), lymphovascular and neural invasion were assessed. IHC was done on all cases using antibodies against Ki-67, cyclin D, E-cadherin, p53, CD31 and each antibody was assessed according to the standard protocol. STATISTICAL ANALYSIS: To calculate the relation between clinical, histopathological parameter, IHC marker and the occurrence of LN metastasis, chi-square test was used. Variables were tested using multivariate logistic regression method to assess the predictive significance. RESULTS: Out of 105 cases studied, 29 cases showed LN metastasis. Maximum numbers of cases affected were females with involvement of buccal mucosa. We found significant association of cervical LN metastasis with high grade of differentiation, lack of E-cadherin expression, high Ki-67 and cyclin D1 expression. In our study; tumor depth, thickness, extent of peritumoral lympho-plasmacytic infiltration, presence of eosinophils, tumor nest type, p53 and microvessel density (MVD) showed no significant correlation. CONCLUSION: Significant association of cervical LN metastasis with high grade of differentiation, lack of E-cadherin expression, high Ki-67 and cyclin D1 expression was seen.

Micronuclei in breast aspirates. Is scoring them helpful?

BACKGROUND: Micronuclei scoring can be used as a biomarker of genotoxic and chromosomal damage. AIMS: 1. To score the spontaneously occurring micronuclei in the baseline population (fibroadenomas) and infiltrating ductal carcinoma, 2. Compare micronuclei frequency in benign tumors and various grades of infiltrating ductal carcinomas. MATERIALS AND METHODS: Study was done at a tertiary hospital where 40 cases of fibroadenoma and 40 cases of infiltrating ductal carcinoma were taken up. May Grunwald Giemsa stained smears were analyzed for micronuclei scoring. STATISTICAL ANALYSIS: Independent sample test (Student t test) was done to look for significant difference occurring between the controls among all grades of infiltrating ductal carcinoma. RESULTS: Mean micronuclei range in fibroadenoma was 1.8 ± 1.9. It was 12.1 ± 9.2, 27.4 ± 27.2 and 100 ± 36.5 in grade I, grade II and grade III carcinomas respectively. CONCLUSION: An increase in micronuclei values was seen from fibroadenoma to infiltrating ductal carcinoma. Micronuclei scoring can be used as a biomarker on fine needle aspiration cytology smears of breast carcinoma.

Expression of vimentin in breast carcinoma, its correlation with Ki67 and other histopathological parameters.

CONTEXT: Vimentin is a mesenchymal marker, known to express in some epithelial carcinomas. AIMS: 1. To find out the expression of vimentin in infiltrating ductal carcinoma of breast (not otherwise specified), 2. To find out the correlation between expression of vimentin and prognostic markers such as tumor size, tumor grade, lymph node status, proliferation index (measured by Ki 67), and Nottingham prognostic index (NPI). MATERIALS AND METHODS: Study was done at Department of Pathology; 50 cases of infiltrating ductal carcinoma (NOS) were studied for tumor grade; immunohistochemistry was done using antibodies against vimentin and Ki 67. Percentages of positive cells were documented. An immunoscore was also calculated for vimentin. Vimentin expression was correlated with tumor size, lymph node status, Nottingham prognostic index, and Ki 67. STATISTICAL ANALYSIS USED: statistical correlation was done using Pearson's chi-square test. A P value less than 0.01 was considered significant. RESULTS: Vimentin expression was seen in 18% of cases. Its expression correlated with high tumor grade and high growth fraction (P value < 0.01). It did not correlate with lymph node status, tumor size, and NPI. CONCLUSIONS: Increased vimentin expression is associated with bad prognostic factors. Immunohistochemistry with vimentin may be helpful in knowing the prognosis in cases of infiltrating ductal carcinoma of breast (NOS).

Malignant peripheral nerve sheath tumor with divergent differentiation.

A malignant peripheral nerve sheath tumor (MPNST) is an uncommon spindle cell sarcoma accounting for approximately 5% of all soft tissue sarcomas. A 55-year-old female with a right suprarenal tumor showed MPNST with additional foci of epithelioid, rhabdomyoblastic, osteogenic and lipogenic differentiation. Although the capacity of MPNST to undergo epithelioid, rhabdomyoblastic, osteogenic and very rarely lipogenic differentiation is reported in literature, the occurrence of all these differentiation in one case has not been described in literature before. To the best of our knowledge, this is only the second MPNST case with lipomatous differentiation.

Subarachnoid spread of germinoma mimicking tuberculous meningitis.

A case of pineal germinaoma spreading along the basal subarachnoid space, clinically and macroscopically at autopsy, resembling tuberculous basal arachnoiditis is reported. Need to carry out CSF cytology of even hemorrhagic CSF is stressed, to diagnose the condition.

Medulloblastoma with extensive nodularity: a variant occurring in the very young-clinicopathological and immunohistochemical study of four cases.

CASE REPORT: Four cases of medulloblastoma with extensive nodularity (MBEN) are described. The patients were 3 years of age or younger at diagnosis. Cranial CT scan disclosed multiple coalescing nodules with peculiar 'grape-like' architecture in three patients. A near total excision of the tumour was performed in all four patients. The patients are currently receiving radiation and chemotherapy. Histologically, the tumours were characterized by extreme nodularity with intranodular uniformity and low proliferative index. The internodular zones were extremely cellular, composed of undifferentiated, mitotically active cells. Bcl-2 protein expression was observed in the cellular zones but was distinctly absent within nodules. DISCUSSION: MBEN represents a variant that occurs in very young children but has a good prognosis. The favourable outcome is probably related to its spontaneous neurocytic differentiation. The pattern of Bcl-2 immunoreactivity in MBENs indicates that this protein could be a key player in the regulation of neuronal differentiation in medulloblastomas.

Unusual case of multiple cellular and malignant schwannomas of the cranial and spinal nerves.

Schwannomas from cranial nerves and spinal roots are most often benign, malignant schwannomas being uncommon in this location. This report describes a unique case of multiple cellular and malignant schwannomas of the cranial and spinal nerves in a patient with features of neurofibromatosis 2. The tumors were arising from left optic, bilateral oculomotor, trochlear, abducent and vestibular nerves, the left facial and the spinal lumbar nerve roots. The tumor arising from the right trochlear nerve was seen excavating into the hippocampus and the left vestibular nerve into the medulla. In addition, the patient had nodular schwannomas adherent to the inferior surface of the optic chiasm and the cerebellum away from the cranial nerves. Hyperplastic schwannosis was noted in perivascular spaces of thalamus, cerebellum and hippocampus on both sides away from the main lesions. The immunohistochemical and ultrastructural profiles of the tumors suggest that neurofibromas and schwannomas are probably not distinct tumors but lie within a spectrum that differs histologically depending on the predominant cell type.

Intracranial haemorrhage resulting from unsuspected choriocarcinoma metastasis.

A retrospective analysis of clinicopathological data of 10 patients with clinically unsuspected cerebral metastatic choriocarcinoma was carried out. All patients were young adult females. History of preceding pregnancy/abortion was forthcoming in 5 cases but none had a prior history of abnormal gestation. Features of raised intracranial tension followed by hemiparesis were the commonest presenting symptoms. A clinicoradiologic diagnosis of intracerebral haemorrhagic mass, either primary or secondary to tumour bleed, cortico-venous thrombosis or arteriovenous malformation, was entertained in 8 out of 10 cases. In other two cases, ring enhancing lesions prompted the diagnosis of granulomatous masses. Eight patients were operated upon, of whom two died after short hospital stay, and were autopsied. All had haemorrhagic masses noted at surgery/autopsy. Accurate diagnosis of metastatic choriocarcinoma was established only by histologic examination of these haemorrhages. This report emphasizes the importance of considering metastatic choriocarcinoma as an important differential diagnosis of haemorrhagic intracerebral lesions in women of child bearing age group. Measurement of serum/CSF level of HCG in suspected cases helps to implement early therapy. The diagnostic value of histopathologic examination of surgically resected blood clots in determining aetiology of intracerebral haemorrhagic masses is highlighted.