Serial Analysis of Gene Expression Identifies Connective Tissue Growth Factor Expression as a Prognostic Biomarker in Gallbladder Cancer

Abstract

Background: Gallbladder cancer (GBC) is an uncommon neoplasm in the United States, but one with high mortality rates.This malignancy remains largely understudied at the molecular level such that few targeted therapies or predictive biomarkers exist. Experimental Design: We built the first series of serial analysis of gene expression (SAGE) libraries from GBC and nonneoplastic gallbladder mucosa, composed of 21-bp long-SAGE tags. SAGE libraries were generated from three stage-matched GBC patients (representing Hispanic/ Latino, Native American, and Caucasian ethnicities, respectively) and one histologically alithiasic gallbladder. Real-time quantitative PCR was done on microdissected epithelium from five matched GBC and corresponding nonneoplastic gallbladder mucosa. Immunohistochemical analysis was done on a panel of 182 archival GBC in high-throughput tissue microarray format. Results: SAGE tags corresponding to connective tissue growth factor (CTGF) transcripts were identified as differentially overexpressed in all pairwise comparisons of GBC (P < 0.001). Realtime quantitative PCR confirmed significant overexpression of CTGF transcripts in microdissected primary GBC (P < 0.05), but not in metastatic GBC, compared with nonneoplastic gallbladder epithelium. By immunohistochemistry, 66 of 182 (36%) GBC had high CTGF antigen labeling, which was significantly associated with better survival on univariate analysis (P = 0.0069, log-rank test). Conclusions: An unbiased analysis of the GBC transcriptome by SAGE has identified CTGF expression as a predictive biomarker of favorable prognosis in this malignancy. The SAGE libraries from GBC and nonneoplastic gallbladder mucosa are publicly available at the Cancer Genome Anatomy Project web site and should facilitate much needed research into this lethal neoplasm.

Long non-coding RNA Malat1 promotes gallbladder cancer development by acting as a molecular sponge to regulate miR-206

“…KRAS mutations are common in GBC [34], and are often linked to resistance to cetuximab, an epithelial growth factor receptor (EGFR) monoclonal antibody [35]. Here we further demonstrate that KRAS and ANXA2 levels are elevated in GBC tissues, in agreement with previous studies [36]. In addition, we find that knockdown of Malat1 inhibits KRAS and ANXA2 expression, and that this can be reversed by co-transfection of miR-206 inhibitor.…”

CTGF is overexpressed in papillary thyroid carcinoma and promotes the growth of papillary thyroid cancer cells

“…Individuals whose gastric carcinoma expressed a high level of CTGF had significantly lower survival rates than those with a low CTGF expression [22]. However, it was noted that patients with lung [23], breast [24], and gallbladder [25] cancers, as well as cartilaginous malignancies [9], whose tumors had high CTGF expression, had significantly better survivals. These data suggest that the role of CTGF in tumor progression depends on the tumor type.…”

Current Management of Gallbladder Carcinoma

“…Transcriptional analysis of GBCs has been accomplished using serial analysis of gene expression profiling. This identified connective tissue growth factor as a prognostic marker as well as a number of other transcripts differentially expressed between GBC and normal gallbladder epithelium [96].…”