Positive Staining For Cytokeratin Research Articles

SMARCA4-Deficient Undifferentiated Carcinoma of the Gastroesophageal Junction: A Case Report

Abstract Introduction/Objective To report a rare case of the newly characterized variant of gastric carcinoma; undifferentiated carcinoma with intact SMARCB1 (INI1) but lost SMARCA4/SMARCA2, and approach to diagnosis. Methods/Case Report A case of gastric mass extending to the gastroesophageal junction, microscopically, showing sheets of atypical, medium to large, discohesive, polygonal cells with large eccentric and pleomorphic nuclei, variably prominent nucleoli, and eosinophilic cytoplasm with brisk mitosis, in the background of marked necrosis and granulomatous inflammation. And having focally positive staining for cytokeratin and negative for melanoma, diffuse large B-cell lymphoma, high grade neuroendocrine tumor, sarcoma, rhabdoid/myoid-specific immunohistochemistry (IHC) markers. The case was further characterized with immunophenotypic, cytogenetic, and molecular findings. Results (if a Case Study enter NA) By immunohistochemistry (IHC), the large tumor cells were positive for vimentin, FLi-1 and patchy CD71. A subset of tumor cells showed CD34 positivity. The tumor cells were mainly negative (with rare focal positivity) for all cytokeratin including (CK AE1/AE3, EMA, and CK OSCAR). Tumor cells were negative for specific markers for melanoma, GIST, sarcomatous tumors, squamous cell carcinoma, hematolymphoid malignancies, and neuroendocrine tumors. EBER-ISH and HHV-8 were negative. The Ki67 showed a proliferative index of ~60%. The tumor cells demonstrated retention of INI1 (SMARCB1) with loss of both SMARCA2 and SMARCA4 by IHC. Molecular studies detected an inactivating mutation of the retinoblastoma-1 gene (RB1 I124Rfs*6) and TP53 with altered function compared to wild-type (TP53-G245S) with no mutations or deletion of SMARCB1 gene. The status of SMARCA4 gene came as indeterminate. No microsatellite Instability detected. Conclusion Our study demonstrates the challenge of diagnosing the SMARCA4/SMARCA1-deficient undifferentiating gastric carcinoma especially with negative cytokeratin stains, showing the immunohistochemistry profile of the tumor and its molecular findings in our case.

Rare Seminal Vesicle Cystadenoma: Case Report and Literature Review

Abstract Introduction/Objective Primary neoplasms of seminal vesicles (SV) are rare, with the majority being metastatic tumors, often from prostatic carcinoma. SV cystadenoma is an exceedingly rare benign epithelial tumor that arises from the remnants of the Mullerian duct. The tumor formed of cysts of variable sizes lined by bland cuboidal epithelium. Herein we present a case of SV cystadenoma in a 38-year-old male, underscoring its rarity and risk of progression to higher grades, as outlined in literature review. Methods/Case Report A 38-year-old male with a history of left atrophic kidney and intermittent pelvic pain due to recurrent obstruction of the SV. Pelvic Magnetic Resonance Imaging revealed a 5.3 cm benign unilocular cyst within the SV, along the posterior aspect of the urinary bladder. The patient underwent successful laparoscopic excision of the left SV. Gross examination of the excised SV demonstrated a 3.5X 2.7X 1.5 cm cyst within the SV. Microscopic examination displayed a benign cyst lined by single to multiple layers of cuboidal epithelium, surrounded by unremarkable SV stroma. The epithelial lining cells exhibited round nuclei with inconspicuous nucleoli, lacking dysplasia. Immunohistochemistry revealed positive staining for cytokeratin 7 and PAX 8 in the epithelial cells, whereas prostatic specific antigen staining was negative. Results (if a Case Study enter NA) NA Conclusion Cystadenoma and mixed epithelial stromal tumors of the SV (MEST) are rare entities, with approximately 30 reported cases. This case demonstrates classic histologic features consistent with low-grade MEST, as per recently proposed grading criteria. Notably, a recent case report has suggested a potential for malignant transformation to high-grade dysplasia and adenocarcinoma in SV cystadenoma, as evidenced by focal epithelial dysplasia.

Clinicopathological and molecular characteristics of microsecretory adenocarcinoma in salivary gland

Objective: To investigate the clinicopathological, immunohistochemical, and molecular genetic characteristics of microsecretory adenocarcinoma (MSA) of the salivary gland, and to improve the understanding of this rare tumor. Methods: Cases originally diagnosed as MSA at the Department of Oral Pathology, the Ninth People's Hospital of Shanghai Jiao Tong University School of Medicine were retrospectively collected. The cases of polymorphous adenocarcinoma and adenocarcinoma, not otherwise specified from January 2000 to January 2020 were reviewed to identify potential misdiagnosed MSA cases. Clinicopathological analysis and follow-up of all confirmed MSA cases were performed, and relevant literature was reviewed. Results: A total of 4 MSA cases were identified, including 2 screened from the polymorphous adenocarcinoma cohort. Of the 4 MSA patients, 3 were male and 1 was female, with an average age of 53 years (range, 37-67 years). Three cases occurred in the palate, and one in the buccal region. The clinical manifestation was usually a slow-growing painless mass. Tumors were generally small, with a maximum diameter ranging from 0.7 to 1.8 cm (average, 1.2 cm). Microscopically, the tumor was unencapsulated and showed an infiltrative growth pattern. The tumor cells appeared small in size and showed bland, cubic and flattened cytological features, forming microcystic lumens and glandular tubes. Significant basophilic secretions were seen in the lumens. Between the tumor nests there was fibro-myxoid stroma. Immunohistochemistry showed diffusely or partially positive staining for cytokeratin 7, S-100, SOX-10, p63 and vimentin and negative staining for p40, mammaglobin, and calponin. The proliferation index of Ki-67 was relatively low (1%-3%). Four MSA cases all harbored SS18 gene rearrangement as shown by fluorescence in situ hybridization (FISH), including 2 cases with MEF2C::SS18 fusion gene through RNA-targeted next generation sequencing. All 4 patients underwent surgical resection without any adjuvant treatments. Three patients were followed up for a period of 2 to 203 months. No tumor recurrence, metastasis, or disease-related death was found. Conclusions: Salivary gland MSA is a novel and rare low-grade carcinoma with unique and consistent histological morphology, immunophenotype, and molecular changes. Immunohistochemical staining and SS18 break apart FISH are useful for the diagnosis of the tumor with atypical morphology and high-grade transformation.

Does Photobiomodulation Affects CK10 and CK14 in Oral Mucositis Radioinduced Repair?

The mechanisms of action of photobiomodulation (PBM) in oral mucositis (OM) are not completely elucidated. To enlighten the role of PBM in the evolution of epithelial maturity in OM ulcers, the present study evaluated the effect of PBM with red (λ) wavelength of 660 nanometers (nm) and infrared of 780 nm in radio-induced OM wounds on the tongue of rats, eight and twenty days after irradiation with single dose of 20 Gy. The percentage area corresponding to positive staining for cytokeratin 10 (CK10) and 14 (CK14) proteins was evaluated in the epithelial area of the lesions, using an immunohistochemical technique (IHC), 8 and 20 days after the induction of lesions, and compared with an untreated control group. CK10 was significantly more expressed in the group treated with 660 nm PBM. CK14 did not show quantitative differences between the groups evaluated. However, whereas in the groups treated with PBM, CK14 was already restricted to the basal layer of the epithelium, as expected in healthy epithelia, in control group it was also expressed in upper layers of the epithelium. In this work, PBM was able to improve epithelial maturity of the repaired OM wound, especially in the 660 nm group.

Long-term and non-invasive in vivo tracking of DiD dye-labeled human hepatic progenitors in chronic liver disease models.

Chronic liver diseases (CLD) are the major public health burden due to the continuous increasing rate of global morbidity and mortality. The inherent limitations of organ transplantation have led to the development of stem cell-based therapy as a supportive and promising therapeutic option. However, identifying the fate of transplanted cells in vivo represents a crucial obstacle. To evaluate the potential applicability of DiD dye as a cell labeling agent for long-term, and non-invasive in vivo tracking of transplanted cells in the liver. Magnetically sorted, epithelial cell adhesion molecule positive (1 × 106 cells/mL) fetal hepatic progenitor cells were labeled with DiD dye and transplanted into the livers of CLD-severe combined immunodeficiency (SCID) mice. Near-infrared (NIR) imaging was performed for in vivo tracking of the DiD-labeled transplanted cells along with colocalization of hepatic markers for up to 80 d. The existence of human cells within mouse livers was identified using Alu polymerase chain reaction and sequencing. NIR fluorescence imaging of CLD-SCID mice showed a positive fluorescence signal of DiD at days 7, 15, 30, 45, 60, and 80 post-transplantation. Furthermore, positive staining of cytokeratin, c-Met, and albumin colocalizing with DiD fluorescence clearly demonstrated that the fluorescent signal of hepatic markers emerged from the DiD-labeled transplanted cells. Recovery of liver function was also observed with serum levels of glutamic-oxaloacetic transaminase, glutamate-pyruvate transaminase, and bilirubin. The detection of human-specific Alu sequence from the transplanted mouse livers provided evidence for the survival of transplanted cells at day 80. DiD-labeling is promising for long-term and non-invasive in vivo cell tracking, and understanding the regenerative mechanisms incurred by the transplanted cells.

Thyroid angiosarcoma: A case report and review of literature.

Thyroid angiosarcoma is a very aggressive and rare malignancy that is mostly seen in elderly of Alpine regions, related to high prevalence of iodine deficient goiter. The major diagnostic challenge is to distinguish between this entity and anaplastic carcinoma and to specify whether it is a primary or metastatic angiosarcoma. The objective of this work is to clarify this distinction through our case report and a revue of literature. We report the case of a 55 years-old female patient, with no medical history, admitted to the General Surgery Department of Ibn Sina Hospital in Rabat for the management of a cervical mass classified sonographically TI-RADS 6. It was also noted a lesion of the skin next to the thyroid lodge. The patient underwent total thyroidectomy with simple post-operative course. A third tumor location was discovered in the stomach during Oesophago-Gastro-Duodenoscopy (OGD). Microscopically, the thyroid, cutaneous and gastric tumors showed rows of oval to spindle-shaped cells, with marked cytonuclear atypia and high mitotic activity. The stroma was richly vascularized and fibro-inflammatory. Immunohistochemical studies demonstrated that the tumor cells showed positive staining for cytokeratin (AE1/AE3), CK7, ERG and CD31, while they exhibited no immunoreactivity for CK20, thyroglobulin or CD34. The first diagnostic challenge is to rule out anaplastic carcinoma. Using and the classification proposed by Cutlan et al. and a large immunohistochemical panel the diagnosis of epithelioid angiosarcoma was retained. Based on epidemiological and clinical criteria, the primary site in our case was more likely to be cutaneous. In summary, thyroid angiosarcoma is an uncommon thyroid neoplasm which poses a problem of differential diagnosis with anaplastic carcinoma and metastatic angiosarcoma. The use of a large panel of antibodies as well as the epidemiological and clinical data can solves the problem in the most difficult cases. • Thyroid angiosarcoma is a very rare malignancy that is mostly seen in elderly of Alpine regions, related to high prevalence of iodine deficient goiter. • It is a very aggressive tumor that can rapidly spread to the cervical lymph nodes, lungs, and brain or can metastasize to the duodenum, small bowel and large bowel. • Metastatic disease is associated with poor prognosis and limits the mean survival time to a few months after diagnosis and surgical treatment. • The first diagnostic challenge is to rule out an angiomatoid anaplastic carcinoma, using a large panel of antibodies. • The second challenge is to specify whether it is a primary or metastatic angiosarcoma especially when other sites are involved, based on clinical and epidemiological criteria.

Constitutive activation of NF-κB inducing kinase (NIK) in the mesenchymal lineage using Osterix (Sp7)- or Fibroblast-specific protein 1 (S100a4)-Cre drives spontaneous soft tissue sarcoma.

Aberrant NF-κB signaling fuels tumor growth in multiple human cancer types including both hematologic and solid malignancies. Chronic elevated alternative NF-κB signaling can be modeled in transgenic mice upon activation of a conditional NF-κB-inducing kinase (NIK) allele lacking the regulatory TRAF3 binding domain (NT3). Here, we report that expression of NT3 in the mesenchymal lineage with Osterix (Osx/Sp7)-Cre or Fibroblast-Specific Protein 1 (FSP1)-Cre caused subcutaneous, soft tissue tumors. These tumors displayed significantly shorter latency and a greater multiple incidence rate in Fsp1-Cre;NT3 compared to Osx-Cre;NT3 mice, regardless of sex. Histological assessment revealed poorly differentiated solid tumors with some spindled patterns, as well as robust RelB immunostaining, confirming activation of alternative NF-κB. Even though NT3 expression also occurs in the osteolineage in Osx-Cre;NT3 mice, we observed no bony lesions. The staining profiles and pattern of Cre expression in the two lines pointed to a mesenchymal tumor origin. Immunohistochemistry revealed that these tumors stain strongly for alpha-smooth muscle actin (αSMA), although vimentin staining was uniform only in Osx-Cre;NT3 tumors. Negative CD45 and S100 immunostains precluded hematopoietic and melanocytic origins, respectively, while positive staining for cytokeratin 19 (CK19), typically associated with epithelia, was found in subpopulations of both tumors. Principal component, differential expression, and gene ontology analyses revealed that NT3 tumors are distinct from normal mesenchymal tissues and are enriched for NF-κB related biological processes. We conclude that constitutive activation of the alternative NF-κB pathway in the mesenchymal lineage drives spontaneous sarcoma and provides a novel mouse model for NF-κB related sarcomas.

Improved filtration method to isolate pure populations of primary bovine endometrial epithelial and stromal cells for immunological studies

ObjectivesIsolation and culture of distinct primary endometrial cells are key to reliable in-vitro models to investigate the uterine immune response and optimse new disease interventions. Details on the isolation method and purity of distinct cell populations is lacking in currently available protocols leading to inconsistent results across laboratories.MethodsBovine endometrial tissue from non-pregnant bovine uteri were collected immediately post-mortem and separated using differential size filtering. Isolations (n = 15) yielded an average of 3.1 × 105 ± 0.7 × 105 epithelial cells and 1.88 × 106 ± 5.44 × 105 stromal fibroblasts per uterine horn. Following expansion in culture, the purity of cell populations was confirmed using morphology and positive staining for cytokeratin and vimentin which identifies epithelial and stromal fibroblast populations, respectively. Using PCR, cDNA from both cell populations was negative for CD45, a marker of immune cells.ResultsOn challenge with a bacterial PAMP (LPS), epithelial and stromal fibroblasts showed a marked increase in the expression of the inflammatory mediators IL8, IL6, S100A8 and S100A9, with both cell populations displaying distinct expression profiles. Here we provide a detailed methodology on the culture of primary bovine endometrial epithelial and stromal cells and demonstrate these cells provide a physiologically relevant model for studies of endometrial inflammation and its regulation.

Laparoscopic posterior pelvic exenteration for primary adenocarcinoma of the rectovaginal septum without associated endometriosis: A case report

The present report describes a case of laparoscopic posterior pelvic exenteration of a primary adenocarcinoma of the rectovaginal septum (PARS) without associated endometriosis. A 49-year-old woman was admitted to hospital for rectal bleeding. Imaging studies showed a 7-cm solid tumor located in the rectovaginal septum, presenting with invasion to the posterior aspect of the uterine cervix and the anterior rectal wall. The patient received laparoscopic posterior exenteration and rectosigmoid anastomosis followed by chemotherapy. There were no intra- or post-operative complications. Histopathological examination of the neoplastic tissue revealed moderate to severe cytological atypia with bizarre multinucleated cells and prominent mitotic figures. Histopathologically, R0 resection was achieved. No endometriotic lesions were confirmed in the primary tumor or other removed tissues. Immunohistochemistry showed positive staining for cytokeratin (CK)7, cancer antigen 125, vimentin, estrogen receptor and p53, but negative staining for CK20, progesterone receptor, p40 and thyroid transcription factor 1. Based on these findings and on the location of the tumor, the neoplasm was diagnosed as PARS without associated endometriosis, which may have arisen from metaplasia of the embryological Müllerian-duct remnants.

Disgerminoma bilateral e hiperplasia endometrial cística com piometra em cadela

Background : : : Ovarian tumors are considered rare both in dogs and cats. Germ cell neoplasms correspond to approximately 15% of the cases and can be classified into dysgerminomas, teratomas and teratocarcinomas. Reports of bilateral dysgerminoma in dogs and cats are rare. The aim of this work was to describe a case of bilateral dysgerminoma and cystic endometrial hyperplasia with pyometra in a bitch by presenting the clinical, surgical, pathological and immunohistochemical findings. Case : A 10-year-old female crossbred dog was presented with a history of vomiting, apathy and purulent vulvar discharge. Physical examination revealed hyperthermia and pain was observed on abdominal palpation. Ultrasound examination revealed cystic formation and increased echogenicity inside the uterus. The animal underwent ovariohysterectomy (OHE). Macroscopically, the ovaries were multinodular, spherical to ovoid in shape, measuring 6 cm in diameter, and with a dark red color; they exhibited a firm consistency and a hemorrhagic cut surface with cystic areas and foci of necrotic tissue. At microscopic evaluation of both ovaries, the neoplasm consisted of large and polyhedral cells with amphophilic cytoplasm which were arranged in cords separated by connective tissue. The cells exhibited an anaplastic appearance, with central nuclei, large amounts of granular chromatin and one or more evident nucleoli. Several mitotic figures, often incomplete and with consequent formation of multinucleated cells similar to giant cells, were observed. Immunohistochemical detection of cytokeratin and vimentin was performed for further examination. The ovarian tumor exhibited positive staining for cytokeratin and negative staining for vimentin. Discussion : Most ovarian tumors are asymptomatic and many times are considered incidental findings during spaying surgeries. On physical examination, affected animals may show palpable abdominal masses, ascites and systemic signs caused by hormonal disturbances. The macroscopic and microscopic findings seen in this case are in accordance with those found in the literature and confirm the diagnosis of bilateral dysgerminoma and cystic endometrial hyperplasia-pyometra. Dysgerminoma should be differentiated from other neoplasias as lymphosarcoma, solid adenocarcinoma, solid granulosa cell tumor, theca cell tumor and teratoma. Diagnosis is performed by histopathological analysis. Immunohistochemistry can be performed to identify the origin of tumoral cells in order to assist the differential diagnosis for other neoplasias. Even though the immunohistochemical result can be positive for cytokeratin, this is not considered usual for dysgerminomas. The positive result for cytokeratin is related to the anaplastic characteristic of the tumor. Most cases of dysgerminomas are described in older dogs and cats, yet it can develop in younger animals. Dysgerminoma is a tumor that is typically unilateral and is hardly ever found in both ovaries. The rarity with which this neoplasia is observed in bitches can be related to the premature age at which OHE is performed.

SINONASAL TERATOCARCINOSARCOMA - A RARE CASE REPORT

Sinonasal teratocarcinosarcoma is a rare and highly malignant tumor characterized by complex admixture of teratomatous and carcinosarcomatous component. Here, we report a case of 72-year-old male presented with complaints of epistaxis, recurrent sinusitis, nasal stuffiness, and nasal blockade for past 5-6 months and his computed tomography report revealed as an inflammatory sinonasal disease with polyposis. Histopathology examination showed fragments of upper respiratory mucosa admixed with fragments of tumor composed of benign and malignant counterparts of both epithelial and mesenchymal components in a variety of histologic patterns. Also noted were the characteristic squamous epithelium with “fetal-type” clear cells, areas of necrosis, mitotic figure and sheets of undifferentiated blastemal like cells. Immunohistochemistry demonstrated positive staining for cytokeratin, epithelial membrane antigen, vimentin, synaptophysin, S-100, chromogranin, and CD99. This polyimmunophenotypic characteristic and diverse histological differentiation provided the basis for the diagnosis. Knowing its highly aggressive behavior, the patient should be treated aggressively and followed up closely. Key words: Carcinosarcom, Fetal epithelium, Polyimmunophenotypic, Teratoid

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OBJECTIVES/SPECIFIC AIMS: To further elucidate the role of estrogen receptor β (ER-β) in the early endometriotic lesion attachment. METHODS/STUDY POPULATION: EECs were immortalized using a telomerase vector. Immortalized cells and parental cells were characterized by genotyping, and expression of ER-β as well as other epithelial cell markers. ER-β was knocked-down in immortalized EECs using lentivirus-mediated shRNA transduction. ER-β knockdown was confirmed by RT-qPCR and Western analysis. EEC cells with or without ER-β knockdown were used to assess their attachment to PMCs in an established in vitro assay (Lucidi, 2005). Results were analyzed with Student t-test. RESULTS/ANTICIPATED RESULTS: Genotyping using karyotype assay confirmed a normal chromosomal profile. Also positive staining for cytokeratin and lack of any staining with vimentin confirms the epithelial origin of these cells. ER-β knockdown has a significant decrease in attachment compared to control (p=0.02). DISCUSSION/SIGNIFICANCE OF IMPACT: Primary and immortalized cells were 46XX, cytokeratin positive, and vimentin negative confirming their epithelial origin. ER-β knockdown has a significant decrease in attachment compared with control.

Prognostic nomogram for patients with hepatocellular carcinoma underwent adjuvant transarterial chemoembolization following curative resection.

Adjuvant transarterial chemoembolization (TACE) is a major option for postoperative hepatocellular carcinoma (HCC) patients with recurrence risk factors. However, individualized predictive models for subgroup of these patients are limited. This study aimed to develop a prognostic nomogram for patients with HCC underwent adjuvant TACE following curative resection.A cohort comprising 144 HCC patients who received adjuvant TACE following curative resection in the Zhongshan Hospital were analyzed. The nomogram was formulated based on independent prognostic indicators for overall survival (OS). The performance of the nomogram was evaluated by the concordance index (C-index), calibration curve, and decision curve analysis (DCA) and compared with the conventional staging systems. The results were validated in an independent cohort of 86 patients with the same inclusion criteria.Serum alpha-fetoprotein (AFP), hyper-sensitive C-reactive protein (hs-CRP), incomplete tumor encapsulation, and double positive staining of Cytokeratin 7 and Cytokeratin 19 on tumor cells were identified as independent predictors for OS. The C-indices of the nomogram for OS prediction in the training cohort and validation cohort were 0.787 (95%CI 0.775-0.799) and 0.714 (95%CI 0.695-0.733), respectively. In both the training and validation cohorts, the calibration plot showed good consistency between the nomogram-predicted and the observed survival. Furthermore, the established nomogram was superior to the conventional staging systems in terms of C-index and clinical net benefit on DCA.The proposed nomogram provided an accurate prediction on risk stratification for HCC patients underwent adjuvant TACE following curative resection.

Abstract P1-01-05: Prognostic values of circulating tumor cells (CTC) and cancer associated macrophage-like cells (CAML) enumerations in metastatic breast cancer: The role for innate immunity in the metastatic process

Abstract Background: The enumeration of circulating tumor cells (CTCs) using the CellSearch assay is a well-established prognostic and predictive marker for metastatic breast cancer (MBC). However, additional prognostic markers are lacking in patients with ≥ 5 CTCs in 7.5 ml of blood. Tumor-associated macrophages (TAMs) are derived from circulating monocytes or tissue-resident macrophages. TAMs have a controversial role in metastasis and anti-tumor processes. Recent studies showed that circulating cancer associated macrophage-like cells (CAMLs) are specialized phagocytic myeloid cells and found in the peripheral blood of patients with solid tumors including breast cancer, but not in healthy individuals. The presence of CAMLs may indicate the activation of innate immunity in cancer patients. The function and prognostic value of CAMLs in MBC is unknown. In the current study, we measured CTCs and CAMLs on the CellSearch™ platform and investigated their prognostic values in MBC. Methods: Peripheral blood samples from 127 stages IV breast cancer patients were collected at baseline before starting first-line therapy. The detection and enumeration of CTCs and CAMLs in 7.5 ml blood sample were performed on the CellSearch™ system. CTCs were identified by cytokeratins (CK-8, 18, and 19) positive and CD45 negative staining. CAMLs were defined by positive staining for cytokeratins and CD45 (Adams et al, PNAS, 111(9):3514-9, 2014). CTCs and CAMLs enumeration in associations with the progression-free survival (PFS) and overall survival (OS) of patients were evaluated using Kaplan Meier curves and Cox proportional hazards modeling. Results: The image review of CAMLs by using CellSearch analysis showed heterogeneous morphological phenotypes. CAMLs are large cells presenting enlarged nuclei or multiple individual nuclei, and both cytokeratin and CD45 positive with diffused cytoplasmic staining. Among the 127 MBC patients, 38 (29.9%) had elevated CTCs (≥5 CTCs), and 21 (16.5%) had at least one CAML detected. Patients with CAMLs had a significantly increased PFS (p=0.0374) and OS (p=0.0042), compared to patients without CAMLs at baseline. Patients with elevated baseline CTCs and CAMLs had worse PFS with a hazard ratio (HR) of 4.04 (95% CI 2.16 -7.56, P<0.0001), compared to patients with < 5 CTCs and without CAMLs. The combined analysis of baseline CTCs enumeration and CAMLs showed similar effect on patient OS. Compared to patients with < 5 CTCs and without CAMLs, patients with < 5 CTCs and with CAMLs, patients with ≥ 5 CTCs and without CAMLs, and patients with ≥ 5 CTCs and with CAMLs, had an increasing trend of death risk, with an HR of 2.66 (95% CI 0.53-13.21), 6.14 (2.10-17.92), and 9.13 (3.05-27.37), respectively (p for trend<0.0001). Conclusion: Baseline enumerations of both individual CTCs and CAMLs are feasible and increase our ability to accurately predict outcome in MBC patients. Evaluation of CAMLs in peripheral blood may be a marker of innate immunity and provide additional prognostic values for MBC. Citation Format: Mu Z, Wang C, Ye Z, Rossi G, Austin L, Yang H, Cristofanilli M. Prognostic values of circulating tumor cells (CTC) and cancer associated macrophage-like cells (CAML) enumerations in metastatic breast cancer: The role for innate immunity in the metastatic process [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-01-05.

An Unusual Presentation of Metastatic Esophageal Cancer

Esophageal cancer is the 6th leading cause of death from cancer and the 5-year survival rate is dismal around 15%-25%. Metastatic spread to the intestinal tract is extremely rare and thus there are no guidelines suggesting preoperative colonoscopy. Here we present a case of esophageal adenocarcinoma which metastasized to the colon, an event that has so far been unreported in the literature. A 70 year old male with a past medical history of poorly differentiated pT3N1 esophageal adenocarcinoma diagnosed in June 2013 presented to the hospital in December 2015 with shortness of breath and weakness for the preceding two weeks. Patient's initial staging workup included CT of his chest, abdomen and pelvis and PET-CT which demonstrated a PET-avid lesion in his left upper lung. He received neoadjuvant chemotherapy with EOF (Epirubicin, Oxaliplatin and Fluorouracil) for five cycles and subsequently underwent both esophagectomy and left upper lobe wedge resection for his malignancies. He declined further adjuvant chemotherapy after his surgery. On admission, CT of the chest revealed a new pulmonary embolus. He was started on a heparin drip and subsequently had two episodes of hematochezia. Given his history of malignancy and new pulmonary embolus upper and lower endoscopy were pursued. EGD revealed a healthy anastomosis at 21 cm from the incisors. Colonoscopy revealed innumerable friable polypoid lesions, some with overlying exudate, measuring 5-30 mm in size found diffusely throughout the colon. Pathology from the biopsies revealed poorly differentiated adenocarcinoma with positive staining for cytokeratin AE1/AE3, CK7 and CDX2 and negative for HER2, CK20, P40, LCA (CD45), neuroendocrine and melanoma markers. Review of his prior pathology from his esophageal tumor demonstrated a matching immunologic profile consistent with the diagnosis of poorly differentiated metastatic esophageal adenocarcinoma. The patient was referred for systemic chemotherapy; however, he unfortunately expired one week after the diagnosis.Figure 1Esophageal adenocarcinoma is becoming more prevalent in the United States and yet remains a very deadly cancer. Despite strict staging protocols preoperatively, we present a case of esophageal adenocarcinoma metastasizing diffusely to the colon. Although rare and a previously unreported condition, it does raise the question of whether colonoscopic surveillance should be included in the staging guidelines for esophageal adenocarcinoma preoperatively.Figure 2

Abstract 4258: Multi-tumor cell culture medium supports a high take rate and improves culture growth rate in five tumor types

Abstract The efficiency of primary cell line derivation is inadequate, which presents a substantial hurdle in the study of many solid tumor types. We examined whether substitution of a novel multi-tumor cell culture medium into existing cell culture protocols may improve the efficiency of primary cell culture and cell line derivation. The multi-tumor medium RETM (Renaissance Essential Tumor Medium) was initially screened for the ability to achieve primary, extended, and continuous cultures from solid tumors of the breast, lung, colon, prostate and ovary (serous and endometrioid). A pan-cytokeratin antibody was then used to confirm epithelial origin of the expanded cell lines. Performance of RETM was compared to two established media types (RPMI, and Fallopian Tube Medium) in extended primary culture of five high-grade serous carcinoma (HGSC) tumor samples isolated from patients’ ascites. Immunofluorescent staining of Pax8, an important Mullerian lineage marker, was used to confirm the purity and the expected lineage identity of the resulting cell lines. To examine genetic drift, we performed SNP genotyping on the original tumor and the cultured cells in our extended primary cultures of HGSC and our continuous cultures from breast, lung, and colon. The resulting genotype was then compared to the original tumor and the percent matching genotype was determined. Of the six tumor types screened, five grew as long term cultures. Without additional optimization of methodology in HGSC, RETM supported a higher take rate (extended culture in 4 of 5 isolates versus 1 of 5 for established media types) as well as a 10-fold higher average growth rate in the same four cases (RETM doublings/day: 0.309 versus 0.030 for RPMI [p = 0.004] and 0.023 for FTM [p = 0.001]). In all cases, fibroblast overgrowth was ruled out by positive cytokeratin staining. The genetic analysis demonstrated a high degree of fidelity in genotypes between the patients’ tumors and the corresponding cell lines as follows: DF30 (HGSC at 23 doublings) 99.7%; DF68 (HGSC at 25 doublings) 95.5%; Wood (breast ductal and lobular carcinoma at 150 doublings) 98.3%; Jacket (lung adenocarcinoma at 150 doublings) 87.2%; and Ferry (colon adenocarcinoma at 30 doublings) 99.6%. We have shown for the first time that a single cell culture medium can support establishment of long-term cell lines established from a number of different tumor types including breast, lung, colon, endometrioid ovarian, and HGSC. Our results demonstrate that, compared to the current standard, RETM significantly improves both culture take rate and growth rate of the extended HGSC primary cultures. Together with the absence of cell culture crisis, the maintenance of genotype across a large number of population doublings indicate these cell lines are genetically stable and may be cultured to higher passages with reduced concern for genetic drift. Citation Format: Elin S. Agoston, Marian Novak, Naghmeh Salimi, Alex Chao, Jeffrey Kent, Agoston T. Agoston, Ronny Drapkin. Multi-tumor cell culture medium supports a high take rate and improves culture growth rate in five tumor types. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4258.

Histologic and immunohistochemical evaluation of mirror image biopsies in oral squamous cell carcinoma

Background and objectiveThe concept of field cancerization has frequently been quoted to explain the occurrence of multiple primary cancers in the head and neck region and recurrence following complete excision of the original tumor. The main objective is to study the occurrence of field changes in mirror image biopsy in relation to histopathological changes in the oral mucosa among oral cancer patients using hematoxylin and eosin and to study the expression pattern of cytokeratin, Ki-67 and p53 in oral squamous cell carcinoma and mirror image biopsy. Materials and methodsA pilot study of 15 patients clinically diagnosed with carcinoma lesion and their corresponding mirror image sites were taken and stained using immunohistochemistry method for the expression of cytokeratin, Ki-67 and p53. ResultsPrimary tumors showed strong positive staining for cytokeratin throughout both the epithelium and malignant epithelial islands but absence of staining for Ki-67 and p53. ConclusionCK may be useful in predicting epithelial differentiation and Ki-67 and p53 act as weak indicators of malignant disease progression in oral tissues.

Synchronous ovarian metastasis from colorectal cancer: A report of two cases

Ovarian metastasis of colorectal cancer is relatively rare. The present study reports two cases of synchronous ovarian metastasis from colorectal cancer, which were managed by cytoreductive surgery. In case one, a 60-year-old female patient presented with a multilocular pelvic tumor and ascites. Virtual colonoscopy revealed a mass in the sigmoid colon; however, no tumor cells were identified on histological examination. Ovarian metastasis from sigmoid colon cancer was suspected and adnexectomy was subsequently performed. Histological examination of the excised tumor revealed adenocarcinoma. Immunohistochemical analysis of the resected tumor revealed positive staining for cytokeratin (CK)20 and caudal-type homeobox 2 (CDX2), and negative staining for CK7, estrogen receptor, progesterone receptor and inhibin. The immunohistological results supported the diagnosis of ovarian metastasis from sigmoid colon cancer. In case two, a 56-year-old female patient presented with a multilocular pelvic tumor and ascites. Colonoscopy identified a rectal tumor, and histological examination revealed moderately-differentiated adenocarcinoma, which was confirmed by cytological analysis of ascites. Subsequently, ovarian metastasis from rectal cancer with peritoneal dissemination was diagnosed, and left ovariectomy and transverse colostomy were performed. Histological examination of the excised tumor revealed moderately-differentiated adenocarcinoma, and immunohistochemical investigation revealed positive staining for CK20 and CDX2, but negative staining for CK7. These immunohistological results indicated ovarian metastasis from rectal cancer. Both patients recovered well and are currently undergoing regular follow-up examinations. The observations from the two cases indicate that ovarian metastases of primary colorectal cancer may present as pelvic tumors and, thus, preoperative examination of the gastrointestinal tract is required. Furthermore, even in cases of widespread colorectal cancer metastases, excision of the ovarian tumor is required to establish a histological diagnosis for the selection of appropriate treatments.

A method for establishing human primary gastric epithelial cell culture from fresh surgical gastric tissues.

At present, biopsy specimens, cancer cell lines and tissues obtained by gastric surgery are used in the study and analysis of gastric cancer, including the molecular mechanisms and proteomics. However, fibroblasts and other tissue components may interfere with these techniques. Therefore, the present study aimed to develop a procedure for the isolation of viable human gastric epithelial cells from gastric surgical tissues. A method was developed to culture human gastric epithelial cells using fresh, surgically excised tissues and was evaluated using immunocytochemistry, periodic acid-Schiff (PAS) staining and cell viability assays. Low cell growth was observed surrounding the gastric tissue on the seventh day of tissue explant culture. Cell growth subsequently increased, and at 12 days post-explant a high number of pure epithelial cells were detected. The gastric cancer cells exhibited rapid growth with a doubling time of 13-52 h, as compared to normal cells, which had a doubling time of 20-53 h. Immunocytochemical analyses of primary gastric cells revealed positive staining for cytokeratin 18 and 19, which indicated that the culture was comprised of pure epithelial cells and contained no fibroblasts. Furthermore, PAS staining demonstrated that the cultured gastric cells produced neutral mucin. Granulin and carbohydrate antigen 724 staining confirmed the purity of gastric cancer and normal cells in culture. This method of cell culture indicated that the gastric cells in primary culture consisted of mucin-secreting gastric epithelial cells, which may be useful for the study of gastric infection with Helicobacter pylori and gastric cancer.

Effect of Human Umbilical Cord Blood Stem Cells on Hepatic Regeneration and Immune Response in Liver Fibrosis in Rats

To study the effect CD34+, CD34- and mononuclear cells in a rat model of liver fibrosis, 65 adult female albino rats were divided into 5 equal groups. Group I served as control, while hepatic fibrosis was induced in the other 4 groups by subcutaneous injection of CCl4 for 8 weeks. Group II did not receive any treatment while groups III, IV and V were treated by CD34+ stem cells, CD34- fraction, or mononuclear cells, respectively, as a single IV injection. Liver function and fibrosis were assessed by serum levels of AST, ALT, GGT, albumin and procollagen III. At the end of the experiments the liver was extracted for immunohistochemical (IHC) detection of homing and immune-response markers, as well as for histopathological study. There was a significant improvement of serum levels of AST, ALT and GGT in the 3 treated groups, whereas serum albumin and procollagen III did not show significant change. Histopathology showed improvement in hepatic damage in the stem cell treated rats. IHC staining showed positive staining for cytokeratin 19, human CD34 and human albumin. IHC staining with monoclonal mouse anti-rat CD68, CD8 and CD4 showed no differences among the 3 groups treated with stem cells. We conclude that HUCB stem cells were transdifferentiated into hepatocytes when transplanted in rats with an injured liver and improved liver functional and structural alterations without stimulating a significant immune response.