Abstract 1963: Characterization of microRNA profilings in tumor tissues and serum from myxofibrosarcoma patients
Authors: Morita Takuya , Fujiwara Tomohiro , Uotani Koji , Yoshida Aki , Ozaki Toshihumi , Kazunori Tsukuda 
Published: July 2016

Abstract
Introduction:
Myxofibrosarcoma (MFS) is a common type of soft tissue sarcoma in elderly patients that exhibits a propensity for local recurrence. The local recurrence rates of MFS was reported to range from 22% to 79% even after wide resection. Despite difficulty in pathological diagnosis and evaluation of residual tumor cells after surgery, there are no useful biomarkers to solve these problems. Emerging reports have indicated the possibility of microRNAs (miRNAs) as promising novel biomarkers in malignant tumors. To date, several miRNAs were shown to be dysregulated in various malignant tissue specimens. However, little has been known about the miRNA dysregulations of tumor tissues as well as blood samples in MFS patients. In this study, we investigated miRNA dysregulations in tumor tissues and serum of MFS patients in order to develop novel biomarkers for diagnosis and monitoring MFS progression.
Methods:
The serum samples were collected from patients treated at our institute, which included 5 MFS patients, 5 age-matched non-MFS patients and 9 healthy controls. The tissue samples were collected from 11 MFS patients and normal tissue (bone, muscle, fat, and nerve). Human MFS cell lines (NMFH-1, NMFH-2), human mesenchymal stem cell (hMSC) and human skeletal muscle cell (SKMC) were also included and used in the validation analysis.
Total RNA was isolated using a miRNeay Serum/plasma Kit (QIAGEN) for serum from MFS patients, and miRNeasy Mini Kit (QIAGEN) for tissue from MFS patients. Serum levels of miRNA in MFS patients were compared with those in non-MFS patients and/or healthy controls by using miRNA microarray analysis, and compared with the results of tissue samples. The miRNA expressions in serum from MFS patients and in culture medium from human MFS cell lines, hMSC and SKMC were profiled by quantitative real-time PCR (qRT-PCR).
Results:
miRNA microarray analysis based on the extracted RNA from tumor tissue and serum collected from MFS patients revealed marked upregulation of five miRNAs in MFS serum samples compared with non-MFS or healthy controls (p &lt; 0.05), which were included as the upregulated miRNAs in MFS tissue specimens. One of six miRNA expression levels in the serum from MFS patients were higher than in that from healthy controls, and also upregulated in the MFS tissues compared with control normal tissues. The expression of the miRNA was higher in all MFS cell lines than in hMSCs and SKMCs. Expression of the miRNA was higher in all MFS cell lines than in hMSCs and SKMCs. The expression of this miRNA in the culture media from MFS cell lines increased with time (24 and 48 hours) and numbers of tumor cells, indicating that the detected miRNA are certainly secreted to the extracellular fluids.
Citation Format: Takuya Morita, Tomohiro Fujiwara, Koji Uotani, Aki Yoshida, Toshihumi Ozaki, Tsukuda Kazunori. Characterization of microRNA profilings in tumor tissues and serum from myxofibrosarcoma patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1963.