Evaluation of p53 and Its Target Gene Expression as Potential Biomarkers of Cholangiocarcinoma in Thai Patients

Abstract

Background: Cholangiocarcinoma (CCA), a common cancer in northeastern Thailand, is a severe disease with poor prognosis and short survival time following diagnosis. DNA damage in CCA is believed to be caused by liver fluke infection in combination with exposure to carcinogens. p53, a tumor suppressor, is the most mutated gene in human cancers including liver fluke-associated CCA. Hence, expression patterns of p53 and its target genes may be useful for diagnosis and/or prognosis of CCA patients. Methods: Differential mRNA expression of p53 and its target genes, namely, FUCA1, ICAM2 MDM2, p21, PAI-1, S100A9, and WIP1 in CCA tissue samples (n = 30) relative to matched adjacent non-tumor tissues was determined by quantitative RT-PCR and compared to clinicopathological features. Level of p53 protein was determined by immunohistochemistry and correlated with the expression of its target genes. Results: Immunohistochemistry showed elevation of p53 protein level in 77% of the cases, while RT-PCR showed downregulation of p53 mRNA and its seven target genes in 23% and 47-97% of the samples. PAI-1 was down-regulated in almost all CCA samples, thus highlighting it as a potential diagnostic marker for CCA. However, no significant clinical associations were found except for down-regulation of WIP1 that was significantly correlated with non-papillary type tissue (p-value = 0.001) and with high p53 protein level (p-value = 0.007). Conclusion: Our results demonstrated statistically significant association between down-regulation of WIP1 with non-papillary type and with high p53 protein level, and PAI-1 was down-regulated in almost all CCA. Therefore, expression level of WIP1 and PAI-1 may be useful for predicting p53 functional status and as a potential diagnostic marker of CCA, respectively.

Upregulated LAMA3 modulates proliferation, adhesion, migration and epithelial‑to‑mesenchymal transition of cholangiocarcinoma cells

“…RNA was extracted from liver tissues of CCA patients by coarsely homogenizing the tissue in TRIzol™ Reagent (Invitrogen) as previously described 43 . The RNA concentration and quality (260/280 ratio) were determined using a Nano Drop UV-spectrophotometer (Thermo Fisher Scientific).…”

Zinc Oxide and Magnesium-Doped Zinc Oxide Nanoparticles Ameliorate Murine Chronic Toxoplasmosis

“…Moreover, Toxoplasma infection regulates many cellular pathways to support its survival and proliferation. Many previous studies reported that the activation of P53 is associated with pathological changes in the cell, such as cell cycle arrest, oxidative stress, and DNA damage [ 8 , 9 , 56 ]. The important role of p53 in parasitic infectious diseases has been reported in previous studies [ 57 , 58 ].…”

“…Two researchers performed the score evaluation. The immunohistochemical score (IHS) of each P53 and CD31 was calculated by multiplying the percentage of positive staining cells by the intensity degree of the stain and ranged from 0 (no immunoreactivity) to 12 (maximum immunoreactivity) [ 56 ]. The staining percentage was classified as follows: 0 = 0%, 1 = <10%, 2 = 10–25%, 3 = 26–50%, and 4 = 51–100%.…”

Diagnostic and Prognostic Value of Circulating Cell-Free DNA for Cholangiocarcinoma

“…A total of 13 somatic mutated genes were identified in plasma ctDNA, including ARID1A, PBRM1, MTOR, FGFR3, TP53, PTEN, NCOR1, EPHA2, PIK3CA, TERT, RASA1, EZH2 and BAP1. Mutations in ARID1A, BAP1, PBRM1, and TP53 were previously reported to be associated with poor prognosis for patients with cancer diseases [47][48][49][50]. ARID1A and PBRM1 encode a subunit of the SWI/SNF complex, and were mostly mutated in CCA, including association with tumor progression [48,49].…”