Shuen-Kuei Liao

Activation of Th1 Immunity Is a Common Immune Mechanism for the Successful Treatment of Hepatitis B and C: Tetramer Assay and Therapeutic Implications

Both chronic hepatitis B and C virus (HBV and HCV) infections respond ineffectively to current antiviral therapies. Recent studies have suggested that treatment outcomes may depend on the development of type 1 T helper (Th1) and Th2 cell responses. Specifically, activation of Th1 immunity may play a major role in successfully treating hepatitis B and C. This model was revisited herein by evaluating immune responses in 36 HBV and 40 HCV patients with or without treatment, in an attempt to find a common immune mechanism for successful treatment. The immune responses in all examined cases were studied by peripheral blood mononuclear cell (PBMC) proliferation and cytokine responses to viral antigens, cytotoxic T lymphocyte (CTL) responses, enzyme-linked immunospot (ELISPOT) assay, and tetramer staining of virus-specific CD8+ T cells. The overall results revealed that all responders among both HBV- and HCV-infected cases displayed significantly higher PBMC proliferation to viral antigens with a predominant Th1 cytokine profile. Furthermore, the Th1-dominant responses were associated with significant enhancement of CTL activities and were correlated with ELISPOT data, while non-responders responded more weakly. During therapy, the numbers of tetramer-staining, virus-specific CD8+ T cells showed greater increases in responders than in non-responders (p = 0.001). The frequencies determined by the tetramer assay were approximately 200-fold higher than data estimated by limiting-dilution analysis. In conclusion, activation of Th1 immunity accompanied by enhancement of CTL activity during therapy is a common immune mechanism for successfully treating hepatitis B and C, and therefore may have important therapeutic implications.

PROSTATIC STROMAL SARCOMA IN A YOUNG ADULT: A CASE REPORT

Prostate stromal sarcoma is quite rare, comprising only 0.1–0.2% of all prostate cancers. Here, we report one case of prostate stromal sarcoma in a 38-year-old man. Initially, the patient suffered from lower urinary tract symptoms, and intravenous pyelography showed a larger filling defect in the bladder. Transrectal ultrasound showed a huge heterogenous mass between the bladder and rectum. Abdominal computed tomography revealed prostate tumor with local invasion. Radical cystoprostatectomy with ileal conduit was performed; pathology revealed high-grade prostate stromal sarcoma with invasion to the right seminal vesicle and urethra. This article describes the pathology and immunohistrochemical features of this case and briefly reviews the literature.

Molecular events associated with epithelial to mesenchymal transition of nasopharyngeal carcinoma cells in the absence of Epstein-Barr virus genome

Epithelial-mesenchymal transition (EMT) is an important process in tumor metastasis. The EMT-related events associated with metastasis of NPC in the absence of EBV have not been elucidated. We established an EBV-negative NPC cell line from a bone marrow biopsy of an NPC patient. Using a Matrigel system we isolated an invasive and non-invasive sublines, designated NPC-BM29 and NPC-BM00. NPC-BM29 acquired an invasive-like phenotype characterized by EMT, marked by down-regulation of E-cadherin and β-catenin with concomitant increased expression of Ets1. NPC-BM29 cells expressed ≥ 10-fold higher of MMP-9 than NPC-BM00 cells. NPC-BM29 cells grew better in 2% serum than NPC-BM00 cells, with a population doubling-time of 26.8 h and 30.7 h, respectively. A marked reduction in colony-formation ability of NPC-BM00 cells compared to NPC-BM29 was observed. Wound-healing assay revealed that NPC-BM29 cells displayed higher motility than NPC-BM00 and the motility was further enhanced by cell treatment with TPA, a PKC activator. Cell surface markers and tumor-associated molecules, AE3, MAK6 and sialyl-Tn, were up-regulated in NPC-BM29 cells, whereas the expression of HLA-DR and CD54 was significantly increased in NPC-BM00 cells. NPC-BM29 consistently released higher levels of IL-8 and IL-10 than NPC-BM00, with low levels of IL-1α expression in both cell lines. Higher level of VEGF production was detected in NPC-BM00 than NPC-BM29 cells. These data show that EBV is not required for exhibiting multiple metastatic phenotypes associated with EMT. More studies that target right molecules/signalings associated with the EMT may offer new therapeutic intervention options for NPC invasion and metastasis.

Identification of genetic alterations in upper urinary tract urothelial carcinoma in end-stage renal disease patients.

Clinical presentations of end‐stage renal disease (ESRD) patients on dialysis with upper urinary tract urothelial carcinoma (UUT‐UC) are different from those with normal renal function. The pathogenesis remains unknown. We investigated the pathogenetic influence of chromosomal aberrations in patient on dialysis with UUT‐UC. The chromosomal aberrations of UUT‐UC specimens from seven dialysis patients were assessed by conventional comparative genomic hybridization (cCGH). Subsequently, we further investigated 20 cases by whole genome and fine‐tiling oligonucleotide array‐based CGH to demonstrate gains and losses, and compared with the clinicopathologic background. The chromosomal aberrations in UUT‐UC specimens from dialysis patients were more complex than in bladder urothelial carcinoma (B‐UC). Our data showed that gains at 5p, 7, 19q, and losses at 4q, 9p, and 15q are common in UUT‐UC of ESRD patients. Gains in regions associated with DNA repair genes were noted in this study. High‐stage and high‐grade tumors displayed more copy number variants. In addition, female ESRD patients with UUT‐UC had more frequent chromosomal aberrations than their male counterparts. In conclusion, unique chromosomal aberrations were indentified in UUT‐UC in ESRD patients.

Total HLA class I loss in a sarcomatoid renal carcinoma cell line caused by the coexistence of distinct mutations in the two encoding β2-microglobulin genes

In renal cell carcinoma (RCC), HLA class I downregulation has been found in about 40% of the lesions examined. Since only scanty information is available about the molecular basis of these defects, we have investigated the mechanism(s) underlying HLA class I antigen downregulation or loss in six RCC cell lines. Five of them express HLA class I antigens although at various levels; on the other hand, HLA class I antigens are not detectable on the remaining cell line, the RCC52 cell line, belonging to a sarcomatoid subtype, even following incubation with IFN-γ. β2-microglobulin (β2m) was not detected in RCC52 cells. Surprisingly, RCC52 cells harbor two mutations in the β2m genes in exon 1: a single G deletion (delG) in codon 6, which introduces a premature stop at codon 7, and a CT dinucleotide deletion (delCT), which leads to a premature stop at codon 55. Analysis of eight clonal sublines isolated from the RCC52 cell line showed that the two β2m gene mutations are carried separately by RCC52 cell subpopulations. The delG/delCT double mutations were detected in two sublines with a fibroblast-like morphology, while the delCT mutation was detected in the remaining six sublines with an epithelial cell morphology. Furthermore, loss of heterozygosity (LOH) of the β2m gene at STR D15S-209 was found only in the epithelioid subpopulation, indicating loss of one copy of chromosome 15. Immunostaining results of the tumor lesion from which the cell line RCC52 was originated were consistent with the phenotyping/molecular findings of the cultured cells. This is the first example of the coexistence of distinct β2m defects in two different tumor subpopulations of a RCC, where loss of one copy of chromosome 15 occurs in one of the subpopulations with total HLA class I antigen loss.

Epstein-Barr virus infection in urothelial transitional cell carcinoma tissues.

To explore a possible correlation of Epstein‐Barr virus (EBV) infection with urothelial tumours, as the mutation of oncogenes, inactivation of tumour suppressor genes and viral infections may be important in the tumorigenesis of urothelial tumours, and EBV has been implicated in the pathogenesis of a variety of lymphoproliferative disorders and several epithelial neoplasms.

Urinary fluorescence in situ hybridization assay for detecting urothelial carcinoma in Taiwanese patients

Study Type – Diagnosis (case series)
Level of Evidence 4

Mixed germ cell tumor metastatic to the skin: case report and literature review.

BackgroundTesticular cancer is the most common cancer for males aged 15~35 years old. The initial presentation is typically an asymptomatic enlarged testicle. The retroperitoneum is the most common metastatic area. Other metastatic sites include the lung, liver, brain, adrenal glands, gastrointestinal tract and spleen. Skin metastasis is a rare event and frequently associated with poor prognosis.Case presentationA 19-year old male was diagnosed testicular mixed germ cell tumor with initial presentation of cutaneous metastasis at scalp and upper abdomen. After radical orchiectomy and four courses of cisplatin-based chemotherapy, the scalp and upper abdominal lesions regressed completely. The size of lung metastases remained unchanged.ConclusionsFor advanced stage testicular cancer, cisplatin-based chemotherapy is still effective to achieve partial response.

Epstein-Barr virus DNA load in tumour tissues correlates with poor differentiation status in non-muscle invasive urothelial carcinomas

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Withaferin A targeting both cancer stem cells and metastatic cancer stem cells in the UP-LN1 carcinoma cell model

Withaferin A targeting both cancer stem cells and metastatic cancer stem cells in the UP-LN1 carcinoma cell model Lai-Lei Ting1, Andy Shau-Bin Chou2, Chin-Hsuan Hsieh3, Shih-Chieh Hsiung3, See-Tong Pang3, Shuen-Kuei Liao4,5 1Department of Radiation Oncology, Cancer Center, Taipei Medical University Hospital, Taipei 110, Taiwan, China. 2Department of Radiology, Tzu-Chi General Hospital, Hualien 970, Taiwan, China. 3Department of Surgery, Division of Uro-Oncology, Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan, China. 4The PhD Program of Cancer Biology and Drug Discovery, Taipei Medical University, Taipei 110, Taiwan, China. 5Department of Research and Development, Vectorite Biomedica Inc., New Taipei City 221, Taiwan, China. Correspondence to: Dr. Shuen-Kuei Liao, The PhD Program of Cancer Biology and Drug Discovery, Taipei Medical University, Taipei 110, Taiwan, China. E-mail: liaosk@h.tmu.edu.tw Aim: As our understanding of cancer stem cell (CSC) biology improves, search for inhibitory agents of CSCs and metastatic CSCs (mCSCs) positive for CXCR4 is warranted. Withaferin A (WA), a withanolide extracted from the medicinal plant Withania somnifera, has been shown to exhibit anti-cancer effects through multiple mechanisms. Whether WA could selectively target CSCs, mCSCs, or nonCSCs of a gastrointestinal (GI) carcinoma tumor remains unclear. Methods: Side-population (SP) analysis, flow cytometric phenotyping and sorting, non-invasive imaging in conjunction with xenotransplantation, and immunohistology were used in this investigation. Results: Using the lymph node metastatic GI cancer cell line UP-LN1, consisting of CD44high/CD24low floating (F) and CD44low/CD24high adherent (A) cell subsets, this study demonstrated that as compared with parental UP-LN1 cells or A cells, WA preferentially reduced F-cell proliferation, tumor sphere formation, and SP cells in vitro in greater effi ciencies by apoptosis. This action was mechanistically mediated via the down-regulation of CXCR4/CXCL12 and STAT3/interleukin-6 axes, both of which are instrumental in the acquisition of metastatic ability. Attenuation of interferon-γ-induced CXCR4 expression in F cells by knockdown with siRNA or blocking with an anti-CXCR4 antibody, followed by Western blot analysis, showed signifi cantly reduced metastatic potential in vitro. The extent of in vitro anti-invasive effect of WA on the IFN-γ-treated F cells was signifi cantly greater than on the F cells without WA treatment, or F cells treated with control siRNA or with control IgG antibody. The observed in vitro effects of WA on the CSC and mCSC targeting were validated by data obtained with non-invasive imaging in NOD/SCID mouse xenotransplantation. Conclusion: WA could effi ciently block the formation of both CSCs and mCSCs in the UP-LN1 cell line, suggesting that WA may be considered an effective therapeutic agent for this type of GI malignancies.