Merkel Cell Polyomavirus Small T Antigen Activates Noncanonical NF-κB Signaling to Promote Tumorigenesis.

Multiple human polyomaviruses (HPyV) can infect the skin, but only Merkel cell polyomavirus (MCPyV) has been implicated in the development of a cancer, Merkel cell carcinoma (MCC). While expression of HPyV6, HPyV7, and MCPyV small T antigens (sT), all induced a senescence-associated secretory phenotype (SASP), MCPyV sT uniquely activated noncanonical NF-κB (ncNF-κB), instead of canonical NF-κB signaling, to evade p53-mediated cellular senescence. Through its large T stabilization domain, MCPyV sT activated ncNF-κB signaling both by inducing H3K4 trimethylation-mediated increases of NFKB2 and RELB transcription and also by promoting NFKB2 stabilization and activation through FBXW7 inhibition. Noncanonical NF-κB signaling was required for SASP cytokine secretion, which promoted the proliferation of MCPyV sT-expressing cells through autocrine signaling. Virus-positive MCC cell lines and tumors showed ncNF-κB pathway activation and SASP gene expression, and the inhibition of ncNF-κB signaling prevented VP-MCC cell growth in vitro and in xenografts. We identify MCPyV sT-induced ncNF-κB signaling as an essential tumorigenic pathway in MCC. IMPLICATIONS: This work is the first to identify the activation of ncNF-κB signaling by any polyomavirus and its critical role in MCC tumorigenesis.

Conventional MR and diffusion-weighted imaging of musculoskeletal soft tissue malignancy: correlation with histologic grading.

AIM: To evaluate proven soft tissue musculoskeletal malignancies blinded to their Fédération Nationale des Centres de Lutte Contre le Cancer histologic grades to identify the predictive values of conventional MR findings and best fit region of interest (ROI) apparent diffusion coefficient (ADC) measurements. MATERIALS AND METHODS: Fifty-one consecutive patients with different histologic grades were evaluated by four readers (R1-4) of different experience levels. Quantitatively, the maximum longitudinal size, tumor to muscle signal intensity ratios, and ADC measurements and, qualitatively, the spatial location of the tumor, its signal alterations, heterogeneity, intralesional hemorrhage or fat, and types of enhancement were assessed. Intraclass correlation, weighted kappa, ANOVA, and Fisher exact tests were used. RESULTS: There were 22/51 (43%) men (mean age ± SD = 52 ± 16 years) and 29/51 (57%) women (mean age ± SD = 54± 17 years), with the majority of tumors 38/51 (75%) in the lower extremities. Histologic grades were I in 8/51 (16%), II in 17/51 (33%), and III in 26/51 (51%), respectively. The longitudinal dimensions were different among three grades (p = 0.0015), largest with grade I. More central enhancements and deep locations were seen in grade III tumors (p = 0.0191, 0.0246). The ADC mean was significantly lower in grade III than in grade I or II (p < 0.0001 and p = 0.04). The ADC min was significantly lower in grade III than in grade I (p = 0.02). Good to excellent agreements were seen for T1/T2 tumor/muscle ratios, longitudinal dimension, and ADC (ICC = 0.60-0.98). CONCLUSION: Longitudinal tumor dimension, central enhancement, and ADC values differentiate histology grades in musculoskeletal soft tissue malignancy with good to excellent inter-reader reliability. KEY POINTS: • The longitudinal tumor dimension of grade III malignancy is smaller than that of grade I (p < 0.0001), and higher-grade tumors are located deeper (p = 0.0246). • The ADC mean is significantly lower in grade III than in grade I or grade II (p < 0.0001 and p = 0.04). • The ADC minimum is significantly lower in grade III than in grade I (p = 0.02).

Fine-needle aspiration cytology for the diagnosis of metastatic melanoma: systematic review and meta-analysis.

OBJECTIVES: To perform a thorough review and meta-analysis of studies that have shown non-image-guided fine-needle aspiration cytology (FNAC) to be highly sensitive and specific for assessing questionable metastatic melanoma to lymph nodes. METHODS: MEDLINE and Scopus were searched for potentially relevant articles with a search string including the words "melanoma" and "fine needle." All relevant articles were screened by two authors (B.J.H. and R.L.S.). Full articles were screened for extractable data, and the data was pooled for analysis. RESULTS: Of 978 unique studies found, 10 (5,518 cases) met our inclusion criteria. In a pooled analysis of palpation and ultrasound-guided fine-needle aspirations, the area under the receiver operating characteristic curve was 0.99 (95% confidence interval [CI], 0.99-1.00). The summary estimates for the sensitivity and specificity were 0.97 (95% CI, 0.95-0.98) and 0.98 (95% CI, 0.98-1.00), respectively. CONCLUSIONS: With a sensitivity and specificity of 0.97 and 0.99, the overall diagnostic accuracy of FNAC for metastatic melanoma is quite high, and with a positive and negative likelihood ratio of 58 and 0.03, FNAC for metastatic melanoma should be the first-line option in a patient with a clinically suspected mass and a history of melanoma.

Preemptive surgery for premalignant foregut lesions.

INTRODUCTION: Preemptive surgery is the prophylactic removal of an organ at high risk for malignant transformation or the resection of a precancerous or "early" malignant neoplasm in an individual with a hereditary predisposition to cancer. Recent advances in molecular diagnostic techniques have improved our understanding of the biologic behavior of these conditions. Predictive testing is an emerging field that attempts to assess the potential risk of cancer development in predisposed individuals. Despite substantial improvement in these forms of testing, all results are imperfect. This information often becomes an important tool that is used by healthcare providers to evaluate the risk-benefit ratio of various risk modifying strategies (i.e., intensive surveillance or preemptive surgery). METHODS: A systematic literature review was performed using Medline and the bibliographies of all referenced publications to identify articles relating to preemptive surgery for premalignant foregut lesions. RESULTS AND DISCUSSION: In this review, we outline the controversies surrounding predictive risk assessment, surveillance strategies, and preemptive surgery in the management of high-grade dysplasia (HGD) in Barrett's esophagus (BE), hereditary diffuse gastric cancer (HDGC), bile duct cysts, primary sclerosing cholangitis (PSC), and pancreatic cystic neoplasms. Resection of BE is supported by the progressive nature of the disease, the risk of occult carcinoma, and the lethality of esophageal cancer. Prophylactic total gastrectomy for HDGC appears reasonable in the absence of accurate screening tests but must be balanced by the impact of surgical complications and altered quality of life. Surgical resection of biliary cysts theoretically eliminates the exposed epithelium to decrease the lifetime risk of cholangiocarcinoma. Liver transplantation for PSC remains controversial given the scarcity of donor organs and inability to accurately identify high-risk individuals. Given the uncertain natural history of pancreatic cystic neoplasms, the merits of selective versus obligatory resection will continue to be debated. CONCLUSIONS: Preemptive operations require optimal judgment and surgical precision to maximize function and enhance survival. Ultimately, balancing the risk of surgical intervention with less invasive interventions or observation must be individualized on a case-by-case basis.

Alteration of Drug Sensitivity in Human Colon Cancer Cells after Exposure to Heat: Implications for Liver Metastasis Therapy using RFA and Chemotherapy.

Radiofrequency ablation (RFA) is gaining popularity for treating colorectal liver metastases by inducing image guided tumor hyperthermia. In order to reduce tumor recurrence, adjuvant therapies have been administered post-RFA. We hypothesized that tumor cells escaping RFA cytotoxicity by being in the sublethal zones of tumor might develop differential behavior toward cytotoxic drugs. Here, we used cultured human colorectal cancer cells to evaluate the interaction between heat treatment and chemotherapeutic agents. Human colon cancer cell lines HT29 and HCT116 were subjected to temperatures of 42 degrees to 50 degrees C for 15 min, in combination with 5-fluorouracil, oxaliplatin, or irinotecan at different sequences. Cytotoxicity was determined by MTT assay. The cell cycle progression was analyzed by flow cytometry with propidium iodide staining. The expression of several genes associated with drug sensitivity was quantitated by real-time RT-PCR before and after heat treatment. Either heat treatment at 45 degrees C by simultaneous or pre-treatment with three different chemotherapeutic agents didn't affect the cytotoxicity of the combined treatment to HT29 and HCT116 cells, except for irinotecan treatment in HCT116 cells. However, when pre-exposure to 45 degrees C, HCT116 cells, but not HT29 cells, developed resistance to these three drugs. In an analysis of cell cycle profile after the drug followed heat treatment, a longer delay in cell cycle progression in HCT116 cells was observed in comparison to HT29 cells. Furthermore, HCT116 and HT29 cells exhibited different expression profiles of several drug-related genes in response to heat treatment at 45 degrees C. An observation of a differential response to the drug and heat treatment sequences between two human colon cancer cell lines suggests that tumor heterogeneity and selection of chemotherapeutic agents need to be under consideration in the clinical setting.

Induction of p21WAF1 expression protects HT29 colon cancer cells from apoptosis induced by cryoinjury.

BACKGROUND: Cryotherapy is a method of in situ destruction of tumors by freeze/thaw mechanisms. Cancer cells located in the peripheral zone of the tumor undergoing cryotherapy can die by apoptosis. We hypothesized that p21WAF1 is involved in the mediation of cryotherapy-induced apoptosis. METHODS: HT29 cells grown on a plate were subjected to -10 degrees C and returned to 37 degrees C for various periods of time. Cells were analyzed by flow cytometry, Western blot, and reverse transcriptase-polymerase chain reaction for determining cell-cycle distribution, p21WAF1 protein expression, and messenger RNA levels, respectively. The p21WAF1 expression in nude mouse tumor xenografts after cryotherapy was examined by immunofluorescence staining. A series of the p21WAF1 promoter cloned into a luciferase reporter vector were transfected into HT29 cells for identifying the response element to cryoinjury. Antisense oligodeoxynucleotide (ODN) was applied to examine the effect of p21WAF1 expression on cryotherapy-induced apoptosis. RESULTS: Both protein and messenger RNA of p21WAF1 were induced by cryoinjury in cultured cells and tumor xenografts. Deletion analysis of the p21WAF1 promoter revealed that a region from -121 to -95 base pairs was responsible for the activation and that this activation was p53 independent. HT29 cells arrested at the G1 phase after cryoinjury. The cryotherapy-induced apoptotic rate in HT29 cells was increased in the presence of antisense p21WAF1 ODN in comparison to the random ODN. CONCLUSIONS: Induction of p21WAF1 increases tumor cell survival and may result in recurrences at treated sites after cryotherapy. Combining antisense ODN targeted against p21WAF1 and cryotherapy may improve clinical outcomes in the treatment of colorectal cancer.

Heat shock protein 70 is induced in mouse human colon tumor xenografts after sublethal radiofrequency ablation.

BACKGROUND: Radiofrequency ablation (RFA) destroys tumor cells by generating high temperatures through ionic vibration. Tumor recurrence may be a direct function of sublethal temperature. Further, a set of proteins called heat shock proteins (HSPs) can be synthesized under heat stress to facilitate recovery of tumor cells from heat damage. METHODS: Subcutaneous xenografts were induced in nude mice by injection with HT29 human colon cancer cells. The tumors were exposed surgically and subjected to RFA. The tumors were randomly assigned to achieve a target tumor temperature of 42 degrees C, 45 degrees C, or 50 degrees C. Total RNA and cell lysates were isolated from tumor tissues and subjected to reverse transcription-polymerase chain reaction and Western blot analyses, respectively, at various time points after treatments for assessing HSP expression. For in vitro experiments, HT29 cells were subjected to variable temperatures, and HSP expression was assayed. RESULTS: During a 50-day follow-up, the recurrence rates were 0% at 50 degrees C, 30% at 45 degrees C, and 100% at 42 degrees C. The messenger RNA and protein levels of HSP90 and HSP27 remained unchanged after RFA at 45 degrees C; however, HSP70 was induced at 4 and 10 hours after RFA. In vitro HT29 culture cells subjected to a heated water bath exhibited a cellular sensitivity to heat and change of HSP expression similar to those in tumor xenografts subjected to RFA. CONCLUSIONS: Our data establish the requisite heat parameters during RFA for human colon tumors in vitro and in vivo. Because HSP70 plays an important role in protecting cell death from a variety of stresses, HSP70 could be a potential target for enhancing the efficacy of RFA.