腫瘤切除之檢體細胞存活度及其萃取之RNA/DNA品質的探討
楊明昕1、 高建璋1、陳進利1、吳勝堂1、查岱龍1
1三軍總醫院 泌尿外科
How is your RNA/DNA quality and sample viability from your resected tumour?
Ming-Hsin Yang1、Chien-Chang Kao1、Chin-Li Chen1、Sheng-Tang Wu1 、Tai-Lung Cha1
1Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
Abstract
Purpose:
Nucleic acid sequencing and tissue viability are frequently used to determine the molecular basis of cancer experiments. Therefore, proper collection of biological specimens is essential to keep the samples viable and inhibit nucleic acid degradation. In this study, we will evaluate the samples we collected from our surgery.
Materials and Methods:
The viability of tumour samples were tested by Flow cytometry. The effect of several parameters, including the method used for tissue lysis, RNA/DNA later treatment, tissue weight at storage, and the time to storage after surgical resection, on the quantity and quality of RNA/DNA extracted was examined. Frozen samples were also evaluated.
Results:
Data showed that the highest quantity of RNA was isolated using a combination of manual and mechanical methods of tissue lysis. Additionally, sectioning the tissues into small pieces and treating them with proper solution prior to storage increased RNA/DNA stability. Tumor samples’ viability can be maintained more than 90% within 30 minutes after we halted the circulation (eg. control the renal artery during partial nephrectomy). Frozen samples can only have 50% viability. Furthermore, high-quality RNA/DNA can only be obtained freshly.
Conclusion:
By providing detail clinical information of the tumour status, the method can be used in preclinical research settings. The application of these results during sample handling and storage in clinical settings will facilitate the genetic diagnosis of cancer and their subsequent treatment.