Squamous cell carcinoma of the lung compared with other histotypes shows higher messenger RNA and protein levels for thymidylate synthase

TLDR

Thymidylate synthase (TS) overexpression is a major mechanism of resistance to antimetabolite chemotherapy in cancer. The study sought to quantify TS mRNA and protein levels in 56 NSCLC resection specimens and assess their association with tumor histology and grade. TS mRNA was measured by real‑time PCR, protein by immunohistochemistry on FFPE sections, and transcriptional activity evaluated by real‑time PCR in snap‑frozen tumor and normal tissues. TS levels were significantly higher in squamous cell carcinoma and high‑grade tumors, with strong correlation between mRNA and protein, indicating TS as a potential biomarker for selecting patients for TS‑inhibiting agents.

Abstract

In patients with cancer, one of the main mechanism of resistance to antimetabolite drugs is related to higher levels of thymidylate synthase (TS) activity.To investigate the association between TS expression and histopathologic data, 56 resection specimens from patients with nonsmall cell lung carcinoma (NSCLC) were collected consecutively. TS messenger RNA (mRNA) was evaluated in tumor specimens by using real-time polymerase chain reaction (PCR) analysis; protein expression was evaluated by using immunohistochemistry (IHC) in formalin-fixed, paraffin-embedded (FFPE) specimens; and the analysis of TS transcriptional regulation activity was performed by using real-time PCR analysis in snap-frozen normal and tumor specimens.The amplification of the TS gene from FFPE tissues was obtained from all samples, with a median level (unit-less ratio) of 1.45 (range, 0.34-5.24); whereas positive TS status at IHC (>10% positive cells) was detected in 56% of samples. It is noteworthy that TS expression was significantly higher in squamous cell carcinoma compared with adenocarcinoma when both mRNA levels (2.17 vs. 1.16; P < .0001) and protein levels (P = .0269) were considered in FFPE specimens, and a strong association was observed between mRNA and protein expression (P = .00017). Moreover, higher TS levels were observed in high-grade tumors (P = .0389 and P = .0068 for mRNA and protein quantification, respectively). The analysis in snap-frozen samples revealed that the TS gene was up-regulated strongly in tumors (P = 3.8 x 10(-12)), and an 8-fold increase (as a cut-off value) in the TS mRNA ratio between tumor and corresponding normal tissue was detected in 32 of 56 patients (57%) bearing preferentially squamous cell tumors (P = .0022) and high-grade tumors (P < .001).Data from the current study consistently indicated higher TS expression levels in squamous cell and in high-grade carcinomas. This information may be useful in selecting which patients with NSCLC should receive treatment with TS-inhibiting agents.