Correction to: Axin gene methylation status correlates with radiosensitivity of lung cancer cells.

Following publication of the original article.

Rare breast metastasis from adenoid cystic carcinoma of the submandibular gland.

Adenoid cystic carcinomas (ACCs) are rare malignant neoplasms of exocrine glands, most commonly found in salivary glands. This report describes a 67-year-old woman with metastatic ACC to the breast, only the third reported case of its kind. The salivary gland ACC was first diagnosed 5 years prior. Routine mammogram identified a Breast Imaging and Reporting Systems (BIRADS) 4 lesion. Core breast biopsy demonstrated findings consistent with metastatic ACC to the breast. The patient ultimately underwent local excision but suffered a recurrence of disease less than 2 months later despite chemotherapy. She passed away 15 months after excision due to complications associated with a small bowel obstruction and decompensated respiratory status from pulmonary metastases. While metastatic salivary ACC to the breast is rare, it is important to be able to distinguish metastatic salivary ACC to the breast from primary ACC of the breast as the treatment considerations for the two disease processes differ significantly.

Axin gene methylation status correlates with radiosensitivity of lung cancer cells.

BACKGROUND: We previously reported that Axin1 (Axin) is down-regulated in many cases of lung cancer, and X-ray irradiation increased Axin expression and inhibited lung cancer cells. The mechanisms, however, were not clear. METHODS: Four lung cancer cell lines were used to detect the methylation status of Axin with or without X-ray treatment. Real-time PCR was used to quantify the expression of Axin, and western blot analysis was applied to measure protein levels of Axin, ß-catenin, Cyclin D1, MMP-7, DNMTS, MeCP2 and acetylated histones. Flow cytometric analysis, colony formation assay, transwell assay and xenograft growth experiment were used to study the biological behavior of the cells with hypermethylated or unmethylated Axin gene after X-ray treatment. RESULTS: Hypermethylated Axin gene was detected in 2 of 4 cell lines, and it correlated inversely with Axin expression. X-ray treatment significantly up-regulated Axin expression in H446 and H157 cells, which possess intrinsic hypermethylation of the Axin gene (P<0.01), but did not show up-regulation in LTE and H460 cells, which have unmethylated Axin gene. 2Gy X-ray significantly reduced colony formation (from 71% to 10.5%) in H157 cells, while the reduction was lower in LTE cells (from 71% to 20%). After X-ray irradiation, xenograft growth was significantly decreased in H157 cells (from 1.15 g to 0.28 g) in comparison with LTE cells (from 1.06 g to 0.65 g). Significantly decreased cell invasiveness and increased apoptosis were also observed in H157 cells treated with X-ray irradiation (P<0.01). Down-regulation of DNMTs and MeCP2 and up-regulation of acetylated histones could be detected in lung cancer cells. CONCLUSIONS: X-ray-induced inhibition of lung cancer cells may be mediated by enhanced expression of Axin via genomic DNA demethylation and histone acetylation. Lung cancer cells with a different methylation status of the Axin gene showed different radiosensitivity, suggesting that the methylation status of the Axin gene may be one important factor to predict radiosensitivity of the tumor.

Histiocytic/dendritic cell transformation of B-cell neoplasms: pathologic evidence of lineage conversion in differentiated hematolymphoid malignancies.

B-cell lymphomas, such as low-grade follicular lymphoma and chronic lymphocytic leukemia/small lymphocytic lymphoma, can transform to histiocytic/dendritic cell sarcoma (H/DS) in rare cases. The diagnosis of this unconventional neoplastic evolution relies on a combination of immunophenotypic analysis and genotypic studies. A genotype identical to that of the primary B-cell neoplasm in a secondary neoplasm with H/DS immunophenotype supports the lineage conversion to H/DS. Putative mechanisms for this unusual phenomenon include dedifferentiation, common immature progenitor, and transdifferentiation models, the latter of which is suggested by clinical laboratory data at the present time. Elucidation of the molecular mechanisms governing this lineage conversion may facilitate the understanding of carcinogenesis of not only hematopoietic but also nonhematolymphoid neoplasms. The clinical outcome of secondary H/DS is dismal, as observed in sporadic cases, and the optimal treatment remains to be determined.

Overexpression of CRKL correlates with poor prognosis and cell proliferation in non-small cell lung cancer.

Crk-Like (CRKL) is an adapter protein that has crucial roles in multiple biological processes, including cell proliferation, adhesion, and migration. Amplification of CRKL gene was found in non-small cell lung cancer (NSCLC). However, the expression pattern of CRKL protein and its clinical significance in human NSCLC have not been well characterized to date. In this study, expression of CRKL was evaluated in 131 NSCLC tissues by immumohistochemistry. CRKL protein was up-regulated in the lung carcinomas compared with adjacent normal lung tissue. Overexpression of CRKL was found in 58 of 131 (44.3%) NSCLC samples and correlated with poor tumor differentiation (P = 0.0042), histological type (adenocarcinoma; P = 0.001), advanced p-TNM stage (P = 0.0004), nodal metastasis (P = 0.0273), high proliferation index (P = 0.0062) and poor overall survival (P = 0.0084). Further univariate and multivariate analysis showed a significant association of CRKL overexpression and worse overall survival in lung cancer patients. In addition, overexpression of CRKL in HBE and H1299 cell lines promoted cell proliferation by facilitating cell cycle progression. Further analysis of cell cycle related molecules showed that CRKL induced cyclin D1, cyclin B1 expression, and increased Rb phosphorylation. In conclusion, this study demonstrated overexpression of CRKL correlated with poor prognosis and lung cancer proliferation by cell cycle regulation.

Systemic mastocytosis with associated clonal hematologic nonmast cell lineage disease: a clinicopathologic review.

Systemic mastocytosis (SM) is a heterogeneous disease with 6 subtypes, including systemic mastocytosis with associated clonal hematologic nonmast cell lineage disease (SM-AHNMD). Bone marrow biopsy specimens show multifocal aggregates of mast cells with predominantly spindle-shaped morphology associated with a myeloid or, less frequently, a lymphoproliferative neoplasm defined by World Health Organization criteria. Neoplastic mast cells abnormally express CD2 and/or CD25, which may be detected by flow cytometry or immunohistochemistry. The pathogenesis of SM-AHNMD is not well understood; however, combined KIT tyrosine kinase receptor mutations and additional genetic events in myeloid stem cells may have a pathogenic role. Reactive mast cell hyperplasia, monocytic/histiocytic proliferations, SM without sufficient criteria for a diagnosis of AHNMD, atypical mast cells associated with PDGFRA rearrangements, and other tryptase-positive myeloid proliferations should be excluded. Overall, the prognosis is poor and largely related to the AHNMD. Cytoreductive therapies, splenectomy, allogeneic bone marrow transplant, and tyrosine kinase inhibitors, excluding imatinib, may have potential efficacy in the treatment of these diseases.

N-terminal 1-54 amino acid sequence and Armadillo repeat domain are indispensable for P120-catenin isoform 1A in regulating E-cadherin.

P120-catenin (p120ctn) exerts important roles in regulating E-cadherin and invasiveness in cancer cells. However, the mechanisms by which p120ctn isoforms 1 and 3 modulate E-cadherin expression are poorly understood. In the current study, HBE, H460, SPC and LTE cell lines were used to examine the effects of p120ctn isoforms 1A and 3A on E-cadherin expression and cell invasiveness. E-cadherin was localized on the cell membrane of HBE and H460 cells, while it was confined to the cytoplasm in SPC and LTE cells. Depletion of endogenous p120ctn resulted in reduced E-cadherin expression; however, p120ctn ablation showed opposite effects on invasiveness in the cell lines by decreasing invasiveness in SPC and LTE cells and increasing it in HBE and H460 cells. Restitution of 120ctn isoform 1A restored E-cadherin on the cell membrane and blocked cell invasiveness in H460 and HBE cells, while it restored cytoplasmic E-cadherin and enhanced cell invasiveness in SPC and LTE cells. P120ctn isoform 3A increased the invasiveness in all four cell lines despite the lack of effect on E-cadherin expression, suggesting a regulatory pathway independent of E-cadherin. Moreover, five p120ctn isoform 1A deletion mutants were constructed and expressed in H460 and SPC cells. The results showed that only the M4 mutant, which contains N-terminal 1-54 amino acids and the Armadillo repeat domain, was functional in regulating E-cadherin and cell invasiveness, as observed in p120ctn isoform 1A. In conclusion, the N-terminal 1-54 amino acid sequence and Armadillo repeat domain of p120ctn isoform 1A are indispensable for regulating E-cadherin protein. P120ctn isoform 1A exerts opposing effects on cell invasiveness, corresponding to the subcellular localization of E-cadherin.

Follicular lymphoma with prominent Dutcher body formation: a pathologic study of 3 cases in comparison with nodal or splenic lymphoplasmacytic lymphoma and marginal zone lymphoma.

Dutcher bodies have been described in lymphoid neoplasms with plasmacytic differentiation, including plasma cell myeloma, lymphoplasmacytic lymphoma, and marginal zone lymphoma. We report a pathologic study of 3 cases of follicular lymphoma with extensive Dutcher body formation in comparison with lymphoplasmacytic lymphoma and marginal zone lymphoma. Of 3 cases, 1 showed a follicular growth pattern, whereas the other 2 cases demonstrated only a vague nodular appearance highlighted by immunohistochemical analysis. Cells containing Dutcher bodies were counted at 25, 90, or 110 per high-power field in each case, respectively. In 2 cases, cells with Dutcher bodies were clustered in an intrafollicular distribution, a possible histopathologic clue for follicular lymphoma. Immunoglobulin M κ was identified as the component of Dutcher bodies in all 3 cases, implying a possible molecular basis for the formation of Dutcher bodies in B-cell lymphomas. All 3 cases had cytogenetic changes supporting the diagnosis of follicular lymphoma, including dual rearrangement of BCL2 and BCL6, rearrangement of BCL2 with trisomy 3 (BCL6), and isolated BCL6 rearrangement. We emphasize immunohistochemical analyses with anti-κ/λ and anti-immunoglobulin heavy-chain isotypes to characterize Dutcher bodies and document clonality in addition to the routine lymphoma workup and indicated cytogenetic studies in B-cell lymphomas with prominent Dutcher bodies.

Expression of ezrin correlates with malignant phenotype of lung cancer, and in vitro knockdown of ezrin reverses the aggressive biological behavior of lung cancer cells.

Ezrin, one of the ezrin-radixin-moesin proteins, is involved in the formation of cell membrane processes such as lamellipodia and filopodia and acts as a membrane-cytoskeleton linker. Its aberrant expression correlates with development and progression of several human cancers. However, the expression of ezrin and its role in lung cancer are currently unknown. In this study, we performed ezrin small interfering RNA transfection in two lung cancer cell lines and examined the effects on malignant phenotypes in cancer cells by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, wound healing, and chamber transwell assays. Ezrin knockdown significantly reduced the proliferation, migration, and invasion of lung cancer cells in vitro. To address the possible mechanisms, we evaluated the expression of adhesion molecules E-cadherin and ß-catenin by Western blot and reverse transcriptase-polymerase chain reaction analyses. The results demonstrated that downregulation of ezrin reduced ß-catenin and increased E-cadherin at the protein level but had no effects on their mRNA levels, suggesting posttranscriptional regulation of these two adhesion molecules. Immunofluorescence assays revealed that ezrin knockdown restored membranous expression of E-cadherin and decreased cytoplasmic ß-catenin in lung cancer cells. In addition, ezrin expression was immunohistochemically evaluated on 135 normal and 183 lung cancer tissues. The expression of ezrin was significantly higher in cancer samples than paired autologous normal lung tissues. In normal bronchial epithelium, ezrin was mainly localized on the apical membrane, while in lung cancers and metastatic foci, ezrin was primarily distributed in cytoplasm. Among lung cancer tissues, expression of ezrin was higher in the invasive front of primary lesions and the highest in lymphatic metastasis. Statistical analysis demonstrated that ezrin expression correlated significantly with lymphatic metastasis and advanced TNM stage. Our data suggest that ezrin may play a crucial role in governing the biological behavior of lung cancer.

Concomitant Waldenstrom macroglobulinemia and IgA plasmablastic myeloma in a patient with untreated IgM paraproteinemia: sequential development of biclonal B-cell neoplasms over a 10-year period in a single individual.

Waldenstrom macroglobulinemia and plasma cell myeloma are both mature B-cell neoplasms primarily involving bone marrow and frequently demonstrating paraproteinemia. Plasma cell myeloma has been described in association with other indolent B-cell lymphomas, particularly chronic lymphocytic leukemia, but concomitant Waldenstrom macroglobulinemia and plasma cell myeloma has not been previously described. We report the first case of sequential development of Waldenstrom macroglobulinemia and plasma cell myeloma over a 10-year period in a 73-year-old man with untreated IgM paraproteinemia. The bone marrow biopsy demonstrated dual populations of lymphoid cells: small mature lymphocytes and immature plasma cells. Although both Waldenstrom macroglobulinemia and plasma cell myeloma were restricted to κ light chain, plasma cell myeloma expressed IgA, whereas the plasmacytic component in Waldenstrom macroglobulinemia produced IgM. The biclonal nature of these 2 B-cell neoplasms was further supported by immunofixation electrophoresis and cytogenetic studies. Although the clinical outcome of plasma cell myeloma when concomitant with other indolent B-cell neoplasms is unfavorable, our patient seemed to respond to high-dose bortezomib plus lenalidomide.

P120-catenin isoforms 1 and 3 regulate proliferation and cell cycle of lung cancer cells via ß-catenin and Kaiso respectively.

BACKGROUND: The different mechanisms involved in p120-catenin (p120ctn) isoforms' 1/3 regulation of cell cycle progression are still not elucidated to date. METHODS AND FINDINGS: We found that both cyclin D1 and cyclin E could be effectively restored by restitution of p120ctn-1A or p120ctn-3A in p120ctn depleted lung cancer cells. When the expression of cyclin D1 was blocked by co-transfection with siRNA-cyclin D1 in p120ctn depleted cells restoring p120ctn-1A or 3A, the expression of cyclin E was slightly decreased, not increased, implying that p120ctn isoforms 1 and 3 cannot up-regulate cyclin E directly but may do so through up-regulation of cyclin D1. Interestingly, overexpression of p120ctn-1A increased ß-catenin and cyclin D1 expression, while co-transfection with siRNA targeting ß-catenin abolishes the effect of p120ctn-1A on up-regulation of cyclin D1, suggesting a role of ß-catenin in mediating p120ctn-1A's regulatory function on cyclin D1 expression. On the other hand, overexpression of p120ctn isoform 3A reduced nuclear Kaiso localization, thus decreasing the binding of Kaiso to KBS on the cyclin D1 promoter and thereby enhancing the expression of cyclin D1 gene by relieving the repressor effect of Kaiso. Because overexpressing NLS-p120ctn-3A (p120ctn-3A nuclear target localization plasmids) or inhibiting nuclear export of p120ctn-3 by Leptomycin B (LMB) caused translocation of Kaiso to the nucleus, it is plausible that the nuclear export of Kaiso is p120ctn-3-dependent. CONCLUSIONS: Our results suggest that p120ctn isoforms 1 and 3 up-regulate cyclin D1, and thereby cyclin E, resulting in the promotion of cell proliferation and cell cycle progression in lung cancer cells probably via different protein mediators, namely, ß-catenin for isoform 1 and Kaiso, a negative transcriptional factor of cyclin D1, for isoform 3.

Sudden death due to undiagnosed T lymphoblastic leukemia/lymphoma in a 5-year-old boy.

Undiagnosed neoplasms in childhood are rare causes of sudden and unexpected death. Deaths due to undiagnosed hematologic malignancies are limited to a small number of case reports. The following case of acute leukemia was diagnosed at forensic autopsy in a 5-year-old boy with no significant past medical history. He complained of nausea and vomiting 2 days before his death, with the subsequent development of fever. Meningitis was the initial suspected cause of death. Findings at autopsy included a 100% cellular bone marrow with greater than 95% blasts. Hemorrhages involving the cerebrum, pons, epicardium, lungs, and thymus were present. Prominent leukemic infiltrates and leukostasis were present in the brain, heart, lungs, spleen, hilar lymph nodes, liver, and kidneys. A peripheral blood smear and automated blood cell count showed a white blood cell count of 435 × 10/L with greater than 80% circulating blasts. Immunohistochemical stains confirmed the diagnosis of T lymphoblastic leukemia/lymphoma. Given these circumstances, the diagnosis of acute leukemia should be considered when an intracerebral hemorrhage and/or visceral hemorrhages are identified on internal examination for appropriate collection of tissue for smears and microscopic examination. This case also highlights the uncommon, although serious, risks associated with acute lymphoblastic leukemia and hyperleukocytosis.

Molecular detection of circulating Sezary cells in patients with mycosis fungoides: could it predict future development of secondary Sezary syndrome? A single-institution experience.

While the majority of patients with early-stage mycosis fungoides (MF) have an excellent prognosis, a few cases progress to secondary Sezary syndrome (sSS), which carries a dismal clinical outcome. We retrospectively analyzed 135 cases of MF/SS and correlated molecular detection of T-cell clones in the skin and blood with other clinicopathologic findings. When stratified by the diagnoses, patients with MF demonstrated a 26.5% (31/117) positive rate for a blood T-cell clone, of which 50% (10/20) had an identical T-cell clone in the skin. Follow-up evaluation showed conversion into sSS or leukemic phase in 50% (5/10) of cases with a positive blood T-cell clone (estimated mean interval 41.8 months) in comparison to no cases in the group without a clone (0/31). Interestingly, 4/5 cases of sSS had an identical T-cell clone in the skin, while the remaining case did not have the test performed on skin for clonal comparison. Kaplan-Meier survival analysis demonstrated a poor clinical outcome in the group with a blood T-cell clone, in comparison with the group without, in overall survival (p < 0.0001) and progression-free survival (p < 0.0001; HR = 22.6). These findings suggest that molecular detection of a blood T-cell clone may have a role in predicting sSS. Due to amplification of non-neoplastic T-cell expansion in a significant number of cases, comparison of blood T-cell clones with skin may have confirmatory value.

A small cell variant of ALK-positive, CD8-positive anaplastic large cell lymphoma with primary subcutaneous presentation mimicking subcutaneous panniculitis-like T-cell lymphoma.

ALK-positive anaplastic large cell lymphoma (ALK+ ALCL) is an uncommon non-Hodgkin's lymphoma of T-cell origin, the majority of which express CD4 and show frequent pan-T-cell antigen loss. While most cases of ALK+ ALCL have the common pattern characterized by anaplastic morphology with hallmark cells, a less common but well-recognized variant with a small cell pattern may pose a diagnostic challenge. We report a case of ALK+ ALCL with small cell morphology and CD8 subset restriction in a 53-year-old male patient who presented primarily with multiple recurrent subcutaneous nodules with histopathologic features simulating a subcutaneous panniculitis-like T-cell lymphoma (SPTCL). The case was initially diagnosed as SPTCL but was reconsidered as ALK+ ALCL when the incidental finding of CD30 positivity on a subsequent biopsy prompted an ALK immunostain, which turned out to be positive in the neoplastic T-cells. The diagnosis of ALK+ ALCL, small cell variant, was then confirmed by detection of an ALK gene rearrangement by FISH analysis. This report highlights a case of ALK+ ALCL with a deceiving clinical and histopathologic presentation, and emphasizes the value of immunohistochemical panel studies and genetic tests in such cases to avoid diagnostic errors.

Recurrent granulosa cell tumor presenting with spontaneous retroperitoneal hemorrhage: A case report.

► Patients diagnosed with granulosa cell tumor require long-term surveillance. ► Recurrent granulosa cell tumor may present as spontaneous retroperitoneal hemorrhage. ► We present an unusual case of recurrent granulosa cell tumor resulting in retroperitoneal hemorrhage.

Primary cutaneous giant cell plasmacytoma in an organ transplant recipient: a rare presentation of a posttransplant lymphoproliferative disorder.

Posttransplant lymphoproliferative disorder (PTLD) is comprised of a spectrum of lymphoid diseases, ranging from early lesions, such as plasmacytic hyperplasia, to monomorphic neoplasms, including plasmacytoma-like lesions. Although PTLD may involve a variety of organs, primary cutaneous PTLD is rare. We report a unique case of Epstein-Barr virus (EBV)-positive primary cutaneous giant cell plasmacytoma developed 5 years after renal/pancreatic transplant in a 55-year-old male patient. The patient received local radiotherapy without reduction in immunosuppression and responded well. A review of the literature identified additional 49 cases of primary cutaneous B-cell PTLD, including 18 cases of plasmacytoma-like lesions. Primary cutaneous B-cell PTLD usually presents years after transplantation, has male preponderance, tends to occur on extremities, is frequently EBV-associated, and predicts a favorable clinical outcome. Unlike PTLD in general, in which EBV-positive cases usually occur earlier than EBV-negative ones, the longer presentation interval in the cutaneous PTLD seems to be uncorrelated to EBV status. Compared with other subtypes of cutaneous B-cell PTLD, plasmacytoma-like lesions have an increased male preponderance and tendency to present on the extremities. Although the majority of cases have been treated with reduction of immunosuppression, antiviral therapy and/or local radiotherapy, and a few with chemotherapy, the best therapeutic intervention for primary cutaneous B-cell PTLD remains to be further investigated with the analysis of more reported cases and large clinical trials.

Primary mediastinal (thymic) large B cell lymphoma with aberrant expression of CD3: a case report with review of the literature.

We report the first case of primary mediastinal large B cell lymphoma (PMBL) with aberrant expression of CD3. PMBL is a subtype of diffuse large B cell lymphoma (DLBCL) and usually presents with bulky mediastinal lesions. Lineage ambiguity/infidelity is uncommon in DLBCL but has been described in sporadic case reports/series. A literature search identifies 13 additional cases of DLBCL expressing CD3, with the majority displaying cytoplasmic expression. Of the 14 total cases, 6 are pyothorax-associated lymphoma, 4 are conventional DLBCL, 2 are plasmablastic lymphoma, one is primary effusion lymphoma and one is PMBL. Two cases show genotypic ambiguity/infidelity with dual clonal IG and TCR gene rearrangements in addition to ambiguous immunophenotypes. Of the 13 cases tested for EBV status, 11 are positive, suggesting an important role of EBV in promoting lineage ambiguity/infidelity. A low threshold for testing EBV status is advocated in DLBCL with phenotypic ambiguity along with panels of immunohistochemical and molecular studies.