Abstract 4597: Telomere maintenance mechanism and expression in colorectal cancer

Abstract

Abstract Tumors can overcome cellular senescence, one of the main barriers to tumor development and immortality by telomere maintenance mechanism (TMM). 90% of colorectal carcinomas (CRC) use telomerase activity (TA) as TMM. Mechanisms of remaining CRC tumors are not well analyzed yet and may utilize alternative lengthening of telomeres (ALT) or a not defined telomere maintenance mechanism (NDTMM). Both TA and expression of telomeric repeat containing RNA (TERRA) were recently identified to correlate with tumor grade. This led us to evaluate possible correlations of TMM and TERRA levels to clinical data. Tissues from 49 CRC patients (35 grade II, 14 grade III, 2 polyps) and cell lines (N = 7) were assessed. Tumor tissues were compared to matched adjacent non-tumor tissues. Mean telomere length (TL) was measured by real-time PCR and telomere restriction fragment (TRF) length analyses. TMM were evaluated by measuring TA with telomere amplification protocol (TRAP) and by detection of ALT with c-circles. Real-time RT-PCR assays for relative quantity (RQ) of transcript levels for TERRA, telomerase genes hTERT and hTERC were determined in relation to levels of house-keeping genes and normal tissues. RQ values were analyzed for significant differences between groups by Mann-Whitney test and were assessed after log-transformation with the Pearson's correlation calculation. TL of grade II and grade III tumors were in the range of 5.5 ± 1.3kbp and 5.3 ± 0.5kbp, respectively (mean ± SD), and were comparable with TL of normal tissue and cell lines. TA was detected with mean of 27 TPG units in 90% (34 of 38) and with mean of 4 units in 24% (9 of 38) of tumor and normal tissues, respectively. All cell lines analyzed demonstrate TA with mean of 51 units. A restricted set of tumors consisting of 2 with and 4 without TA was analyzed for ALT. One of the analyzed tumors without detectable TA showed significantly elevated levels of c-circles, but did not reach levels of sarcomas and cell models with ALT, indicating the existence of NDTMM in CRC. Within the transcripts analyzed, TERRA levels showed a moderate negative correlation with TA (Pearson r=−0.45, p<0.0001), supporting the inhibitory function of TERRA on TA as recently described in vitro. TERRA levels of grade II and III tumors decreased compared to normal tissues, but did not differ between grades. However, TA and TERRA levels of tumors increased and decreased, respectively, in stage II compared to stage I tumors, indicating a role in tumor progression. Correlation with patient survival will be performed and presented. In conclusion, we identified TA and NDTMM, but not ALT in our patient series of CRC. Our preliminary data suggest that TERRA expression together with TA correlates with diagnoses and thus can be considered promising candidates as clinical markers for CRC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4597. doi:1538-7445.AM2012-4597