Impact of ischemia and procurement conditions on gene expression in renal cell carcinoma.
Impact of ischemia and procurement conditions on gene expression in renal cell carcinoma.
复制标题
DOI：
10.1158/1078-0432.ccr-12-2606
复制
发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Bratslavsky G
中科院分区：
文献类型：
--
作者：
Liu NW;Sanford T;Srinivasan R;Liu JL;Khurana K;Aprelikova O;Valero V;Bechert C;Worrell R;Pinto PA;Yang Y;Merino M;Linehan WM;Bratslavsky G
先前的研究表明，缺血改变了正常和恶性组织中的基因表达。没有研究评估缺血对肾肿瘤的影响。本研究探讨缺血和组织采购条件对肾细胞癌RNA完整性和基因表达的影响。对10例肾透明细胞癌患者施行了10例肾肿瘤切除术。肿瘤切除后立即将一块肿瘤快速冷冻。将剩余的肿瘤样品储存在4 ° C、22 ° C和37 ° C下，并在5、30、60、120和240分钟时冷冻。对所有组织样本进行组织学评价，仅选择肿瘤大于80%的组织样本进行进一步分析。通过电泳图确认RNA完整性，并使用RIN指数定量。通过在零时间点和从相同肿瘤获得的来自各种条件的等分试样之间的配对双样本t检验来评估改变的基因表达。进行了140个微阵列。在37 ℃下切除后240分钟观察到一些RNA降解。超过4，000个基因的表达因缺血时间或储存条件而显着改变。最大的基因表达变化观察到较长的缺血时间和较温暖的组织采购条件。来自肾癌的RNA在手术切除后保持完整长达4小时，无论储存条件如何。尽管RNA保存良好，切除后的时间和采购条件显着影响基因表达谱。对采集前变量的密切关注对于准确的肿瘤分析至关重要。
Previous studies have shown that ischemia alters gene expression in normal and malignant tissues. There are no studies that evaluated effects of ischemia in renal tumors. This study examines the impact of ischemia and tissue procurement conditions on RNA integrity and gene expression in renal cell carcinoma. Ten renal tumors were resected without renal hilar clamping from 10 patients with renal clear cell carcinoma. Immediately after tumor resection, a piece of tumor was snap frozen. Remaining tumor samples were stored at 4C, 22C and 37C and frozen at 5, 30, 60, 120, and 240 minutes. Histopathologic evaluation was performed on all tissue samples, and only those with greater than 80% tumor were selected for further analysis. RNA integrity was confirmed by electropherograms and quantitated using RIN index. Altered gene expression was assessed by paired, two-sample t-test between the zero time point and aliquots from various conditions obtained from the same tumor. One hundred and forty microarrays were performed. Some RNA degradation was observed 240 mins after resection at 37C. The expression of over 4,000 genes was significantly altered by ischemia times or storage conditions. The greatest gene expression changes were observed with longer ischemia time and warmer tissue procurement conditions. RNA from kidney cancer remains intact for up to 4 hours post surgical resection regardless of storage conditions. Despite excellent RNA preservation, time after resection and procurement conditions significantly influence gene expression profiles. Meticulous attention to pre-acquisition variables is of paramount importance for accurate tumor profiling.
肝细胞生长因子刺激的肾小管有丝分裂：对C-MYC，C-FOS，C-MET，VEGF和VHL肿瘤抑制剂和相关基因的表达的影响。
影响因子：
8.8
作者：
Clifford, S C;Czapla, K;Richards, F M;O'Donoghue, D J;Maher, E R
通讯作者：
Maher, E R