Chi-Jung Huang

Associations between VHL genotype and clinical phenotype in familial von Hippel–Lindau disease

Background  Von Hippel–Lindau (VHL) disease is an autosomal dominant hereditary disorder associated with tumours and cysts in the central nervous system (CNS) and other visceral organs. Germline mutations in the VHL gene on chromosome 3p25–26 are considered the cause of this disease.

Ribosomal Protein S27-Like in Colorectal Cancer: A Candidate for Predicting Prognoses

Background The development and progression of colorectal cancer (CRC) involve a complex process of multiple genetic changes. Tumor suppressor p53 is capable of determining the fate of CRC cells. However, the role of a p53-inducible modulator, ribosomal protein S27-like (RPS27L), in CRC is unknown. Methods Here, the differential expression of RPS27L was examined in the feces and colonic tissues of CRC patients, to explore its possible correlation with patient survival and to investigate the cellular mechanisms underlying their clinical outcomes. Eighty intermediate-stage CRC patients (42 at stage II and 38 at stage III) were divided into two groups according to their fecal RPS27L mRNA levels. The survival probabilities of the groups were estimated using the Kaplan–Meier method. The RPS27L protein in the colonic tissues of stage III patients with different prognoses was further examined immunohistochemically. RPS27L expression in LoVo cells was manipulated to examine the possible cellular responses in vitro. Results Elevated RPS27L expression, in either feces or tissues, was related to a better prognosis. In vitro, RPS27L-expressing LoVo cells ceased DNA synthesis and apoptotic activity while the expression of their DNA repair molecules was upregulated. Conclusions Elevated RPS27L may improve the prognoses of certain CRC patients by enhancing the DNA repair capacity of their colonic cells, and can be determined in feces. By integrating clinical, molecular, and cellular data, our study demonstrates that fecal RPS27L may be a useful index for predicting prognoses and guiding personalized therapeutic strategies, especially in patients with intermediate-stage CRC.

Prognostic Significance of C-Reactive Protein Polymorphism and KRAS/BRAF in Synchronous Liver Metastasis from Colorectal Cancer

Background The liver is the most common target organ in the metastasis of colorectal cancer (CRC). Synchronous liver metastases may confer a poorer prognosis than metachronous metastases, and genetic alterations and an inflammatory response have also been associated with a poor prognosis in cases of a liver metastasis arising from CRC. However, few studies have examined the relationship between KRAS mutations and inflammatory status in CRC, especially with respect to liver metastases. Methods The effect of the activated mitogen-activated protein kinase pathway and another protein involved in inflammation, C-reactive protein, in liver metastases were examined. We aimed to determine the impact of the CRP-specific single nucleotide polymorphism (SNP) rs7553007 in liver metastasis on the CRC-specific survival (CSS) of patients after colorectal liver metastasectomy. Results We found no significant differences in genotype distributions and allele frequencies at the CRP SNP rs7553007 between CRC patients with liver metastasis and the control group. CSS rates were low in the subgroup of patients with synchronous metastasis with the A-allele (A/A and A/G) at rs7553007 or mutated KRAS/BRAF in liver metastatic specimens. Furthermore, the CRP SNP rs7553007 (hazard ratio [HR] = 1.101; 95% confidence interval [CI] = 1.011–1.200; P = 0.027) and KRAS/BRAF mutations (HR = 2.377; 95% CI = 1.293–4.368; P = 0.005) remained predictive for the CSS of CRC patients with synchronous liver metastasis in multivariate analysis. Conclusions Both the CRP SNP rs7553007 and KRAS/BRAF mutations were independent prognostic factors for CRC patients with synchronous liver metastasis.

Lowly Expressed Ribosomal Protein S19 in the Feces of Patients with Colorectal Cancer

Colorectal cancer (CRC) has become one of the most common fatal cancers. CRC tumorigenesis is a complex process involving multiple genetic changes to several sequential mutations or molecular alterations. P53 is one of the most significant genes; its mutations account for more than half of all CRC. Therefore, understanding the cellular genes that are directly or indirectly related to p53 is particularly crucial for investigating CRC tumorigenesis. In this study, a p53-related ribosomal protein, ribosomal protein S19 (RPS19), obtained from the feces of CRC patients is evaluated by using specifically quantitative real-time PCR and knocked down in the colonic cell line by gene silencing. This study found that CRC patients with higher expressions of RPS19 in their feces had a better prognosis and consistent expressions of RPS19 and BAX in their colonic cells. In conclusion, the potential mechanism of RPS19 in CRC possibly involves cellular apoptosis through the BAX/p53 pathway, and the levels of fecal RPS19 may function as a prognostic predictor for CRC patients.

Hydrostatic pressure enhances mitomycin C induced apoptosis in urothelial carcinoma cells

OBJECTIVES Urothelial carcinoma (UC) of the bladder is the second most common cancer of the genitourinary system. Clinical UC treatment usually involves transurethral resection of the bladder tumor followed by adjuvant intravesical immunotherapy or chemotherapy to prevent recurrence. Intravesical chemotherapy induces fewer side effects than immunotherapy but is less effective at preventing tumor recurrence. Improvement to intravesical chemotherapy is, therefore, needed. METHODS AND MATERIALS Cellular effects of mitomycin C (MMC) and hydrostatic pressure on UC BFTC905 cells were assessed. The viability of the UC cells was determined using cellular proliferation assay. Changes in apoptotic function were evaluated by caspase 3/7 activities, expression of FasL, and loss of mitochondrial membrane potential. RESULTS Reduced cell viability was associated with increasing hydrostatic pressure. Caspase 3/7 activities were increased following treatment of the UC cells with MMC or hydrostatic pressure. In combination with 10 kPa hydrostatic pressure, MMC treatment induced increasing FasL expression. The mitochondria of UC cells displayed increasingly impaired membrane potentials following a combined treatment with 10 μg/ml MMC and 10 kPa hydrostatic pressure. CONCLUSIONS Both MMC and hydrostatic pressure can induce apoptosis in UC cells through an extrinsic pathway. Hydrostatic pressure specifically increases MMC-induced apoptosis and might minimize the side effects of the chemotherapy by reducing the concentration of the chemical agent. This study provides a new and alternative approach for treatment of patients with UC following transurethral resection of the bladder tumor.

A vitronectin M381T polymorphism increases risk of hemangioblastoma in patients with VHL gene defect

Hemangioblastomas, highly vascular tumors, occur sporadically or associated with von Hippel–Lindau (VHL) disease. Diverse mutations in the VHL gene inactivate the VHL protein and constitute the molecular etiology of the disease. Changes in VHL gene were analyzed in patients with multiplex ligation-dependent probe amplification and single-strand conformation polymorphism analyses. We report here that other angiogenesis-related changes in vitronectin were identified with 2D electrophoresis of plasma samples and restriction fragment length polymorphisms. Our findings revealed that most patients (80.0%) with a familial VHL deletion carried the threonine (T) allele at vitronectin codon 381. Adults simultaneously carrying a VHL defect and the T allele were 5.0-fold more likely to be affected by VHL disease than were methionine/methionine (M/M) homozygotes carrying a VHL defect. Patients with sporadic hemangioblastoma, C-terminally truncated VHL protein or a large deletion in the VHL gene, and the T allele were 18.0-fold more likely to develop recurrent disease. Taken together, individuals with mutated VHL are more likely to be affected by familial or recurrent sporadic hemangioblastoma when carrying the M/T or T/T genotype at codon 381 of vitronectin.

Comparative Proteomic Analysis of Placenta-Derived Multipotent Cells (PDMCs) and Its Differentiated Neuron

Placenta-derived multipotent cells (PDMCs) are defined as a group of cells with differentiating and self-renewal properties. The proteomic experimental approach is analyzing a proteome in a high-through-put way. The field of proteomics has several technologies including two-dimensional gel eletrophoresis (2-DE) for protein separation and quantification, mass spectrometry-based protein identification and bioinformatics that integrated technologies. We used IBMX to induce the differentiation of PDMCs to neuron. Further, those cells would serve as a model to apply proteomics strategy to global profile the proteome of PDMCs and its differentiated neuron. We also compared the up- and down-regulated proteins during PDMCs differentiation trying to find the key bio-factors behind PDMCs differentiated into neuron. We believe those findings will give as several candidates for key players in the neuron differentiation. It will help us decipher the mechanism of neuron differentiation and provide us a new version of disease treatment.

Serum CA125 concentration as a predictor of peritoneal dissemination of colorectal cancer in men and women

AbstractPeritoneal dissemination (PD) of cancer is difficult to diagnose. Previous reports have shown that carbohydrate antigen 125 (CA125) is a sensitive marker of PD of gastric cancer. However, CA125 has not been evaluated as a marker of colorectal cancer (CRC), and its accuracy in men is controversial. The aim of this study was to compare the ability of CA125 and carcinoembryonic antigen (CEA) to predict PD of CRC in men and women.Preoperative CA125 and CEA concentrations were measured in 853 people (510 men, 343 women) over 10 years. PD was confirmed intraoperatively in 57 patients. The predictive ability was compared between CA125 and CEA.Compared with CEA, CA125 concentration had a lower sensitivity, higher specificity, and diagnostic accuracy, and significantly greater area under the curve. Further analysis of CA125s sensitivity and specificity among CEA-negative group (n = 514) showed acceptable sensitivity (57.1%) and good specificity (92.0%). In men and women, CA125 concentration did not increase with stage from I to IV unless PD was present (P < 0.001). CEA concentration was increased in women with metastasis with PD (P < 0.001) or without PD (P < 0.001), but was increased only in men with metastasis without PD (P < 0.01). CA125 concentration correlated with PD grade for men and women, but CEA concentration correlated with grade only in women.When analyzed according to the primary tumor site, CA125 concentration in men did not differ between patients with the primary site in the right or left colon, or the rectum, regardless of PD status. By contrast, CA125 concentration differed between PD-positive and PD-negative patients with cancer in the right (P < 0.001) or left (P < 0.001) colon but not in the rectum. CEA concentration in men did not differ according to the primary site or PD status. In women, CA125 and CEA concentrations differed significantly between the PD-positive and PD-negative groups in patients with the primary tumor in the right (P < 0.001) or left (P < 0.001) colon; tumor sites did not differ between the PD-positive and PD-negative groups.These findings suggest that CA125 is a better tumor marker than CEA for predicting PD of CRC in both men and women.