Ming-Hong Tsai1,2, Wen-Chi Chen3, Sung-Liang Yu3,4, Chun-Chieh Chen3, Tzu-Ming Jao3, Chi-Yen Huang3, Sheng-Tai Tzeng3, Sou-Jhy Yen1, Ya-Chien Yang5,6. 1. Department of Surgery, Cardinal Tien Hospital, New Taipei City, Taiwan. 2. School of Medicine, Fu-Jen Catholic University, New Taipei City, Taiwan. 3. Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, Taipei, Taiwan. 4. Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan. 5. Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, Taipei, Taiwan. ycyangntu@ntu.edu.tw. 6. Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan. ycyangntu@ntu.edu.tw.
Abstract
BACKGROUND: Shisa3 is a novel tumor suppressor identified in lung cancer. However, its antitumor activity in other human cancers and the mechanism of gene inactivation remain unknown. METHODS: SHISA3 expression was measured by reverse transcription-PCR (RT-PCR) and quantitative RT-PCR (RT-qPCR). DNA methylation was determined by bisulfite sequencing and pyrosequencing. RESULTS: Down-regulation of SHISA3 expression was observed in all of 11 colorectal cancer (CRC) cell lines and was further confirmed in 34 (65.4 %) of 52 colorectal carcinomas by RT-qPCR. Four of six CRC cell lines could restore SHISA3 expression after treatment with 5-aza-2'-deoxycytidine. Tumor-specific methylation of five CpG sites in the first intron of SHISA3 was identified by bisulfite sequencing, and their methylation levels were quantified in 127 pairs of primary CRC tissues by bisulfite pyrosequencing. The methylation levels of SHISA3 in tumors were noticeably higher than that in their matched normal mucosae. In addition, SHISA3 hypermethylation was significantly associated with an increased risk of disease recurrence in patients with stage II and III disease (P = 0.007) and was an independent predictor of poor overall survival [hazard ratio (HR) 2.9, 95 % confidence interval (CI) 1.5-5.8; P = 0.002] and disease-free survival (HR 4.0, 95 % CI 1.6-10.2; P = 0.003) of CRC patients. CONCLUSIONS: SHISA3 gene is epigenetically inactivated in a substantial fraction of CRC, and its hypermethylation is of prognostic significance in predicting clinical outcome. The quantitative bisulfite pyrosequencing assay established could be a cost-effective tool for providing a potential biomarker of adverse prognosis in CRC.
BACKGROUND:Shisa3 is a novel tumor suppressor identified in lung cancer. However, its antitumor activity in other humancancers and the mechanism of gene inactivation remain unknown. METHODS:SHISA3 expression was measured by reverse transcription-PCR (RT-PCR) and quantitative RT-PCR (RT-qPCR). DNA methylation was determined by bisulfite sequencing and pyrosequencing. RESULTS: Down-regulation of SHISA3 expression was observed in all of 11 colorectal cancer (CRC) cell lines and was further confirmed in 34 (65.4 %) of 52 colorectal carcinomas by RT-qPCR. Four of six CRC cell lines could restore SHISA3 expression after treatment with 5-aza-2'-deoxycytidine. Tumor-specific methylation of five CpG sites in the first intron of SHISA3 was identified by bisulfite sequencing, and their methylation levels were quantified in 127 pairs of primary CRC tissues by bisulfite pyrosequencing. The methylation levels of SHISA3 in tumors were noticeably higher than that in their matched normal mucosae. In addition, SHISA3 hypermethylation was significantly associated with an increased risk of disease recurrence in patients with stage II and III disease (P = 0.007) and was an independent predictor of poor overall survival [hazard ratio (HR) 2.9, 95 % confidence interval (CI) 1.5-5.8; P = 0.002] and disease-free survival (HR 4.0, 95 % CI 1.6-10.2; P = 0.003) of CRC patients. CONCLUSIONS:SHISA3 gene is epigenetically inactivated in a substantial fraction of CRC, and its hypermethylation is of prognostic significance in predicting clinical outcome. The quantitative bisulfite pyrosequencing assay established could be a cost-effective tool for providing a potential biomarker of adverse prognosis in CRC.
Authors: Muriel X G Draht; Danny Goudkade; Alexander Koch; Heike I Grabsch; Matty P Weijenberg; Manon van Engeland; Veerle Melotte; Kim M Smits Journal: Clin Epigenetics Date: 2018-03-14 Impact factor: 6.551
Authors: Andrea Rossnerova; Katerina Honkova; Daniela Pelclova; Vladimir Zdimal; Jaroslav A Hubacek; Irena Chvojkova; Kristyna Vrbova; Pavel Rossner; Jan Topinka; Stepanka Vlckova; Zdenka Fenclova; Lucie Lischkova; Pavlina Klusackova; Jaroslav Schwarz; Jakub Ondracek; Lucie Ondrackova; Martin Kostejn; Jiri Klema; Stepanka Dvorackova Journal: Int J Mol Sci Date: 2020-03-31 Impact factor: 5.923