Endogenous reference RNAs for microRNA quantitation in formalin-fixed, paraffin-embedded lymph node tissue

被引：12

作者：

Inada, Katsushige ^{[1
]}

Okoshi, Yasushi ^{[1
,2
]}

Cho-Isoda, Yukiko ^{[3
]}

Ishiguro, Shingo ^{[3
]}

Suzuki, Hisashi ^{[2
,4
]}

Oki, Akinori ^{[2
,5
]}

Tamaki, Yoshio ^{[2
,6
]}

Shimazui, Toru ^{[2
,7
]}

Saito, Hitoaki ^{[8
]}

Hori, Mitsuo ^{[1
]}

Iijima, Tatsuo ^{[8
]}

Kojima, Hiroshi ^{[2
,3
]}

机构：

[1] Ibaraki Cent Hosp, Dept Hematol, Ibaraki, Japan

[2] Univ Tsukuba, Ibaraki Clin Educ & Training Ctr, Fac Med, Ibaraki, Japan

[3] Ibaraki Cent Hosp, Dept Med Oncol, Ibaraki, Japan

[4] Ibaraki Cent Hosp, Dept Thorac Surg, Ibaraki, Japan

[5] Ibaraki Cent Hosp, Dept Obstet & Gynecol, Ibaraki, Japan

[6] Ibaraki Cent Hosp, Dept Radiat Oncol, Ibaraki, Japan

[7] Ibaraki Cent Hosp, Dept Urol, Ibaraki, Japan

[8] Ibaraki Cent Hosp, Dept Pathol, Ibaraki, Japan

来源：

SCIENTIFIC REPORTS | 2018年 / 8卷

基金：

日本学术振兴会;

关键词：

MIRNA EXPRESSION ANALYSIS; PCR DATA; NORMALIZATION; GENES; IDENTIFICATION; TARGETS;

D O I：

10.1038/s41598-018-24338-7

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Lymph node metastasis is one of the most important factors for tumor dissemination. Quantifying microRNA (miRNA) expression using real-time PCR in formalin-fixed, paraffin-embedded (FFPE) lymph node can provide valuable information regarding the biological research for cancer metastasis. However, a universal endogenous reference gene has not been identified in FFPE lymph node. This study aimed to identify suitable endogenous reference genes for miRNA expression analysis in FFPE lymph node. FFPE lymph nodes were obtained from 41 metastatic cancer and from 16 non-cancerous tissues. We selected 10 miRNAs as endogenous reference gene candidates using the global mean method. The stability of candidate genes was assessed by the following four statistical tools: BestKeeper, geNorm, NormFinder, and the comparative Delta Ct method. miR-103a was the most stable gene among candidate genes. However, the use of a single miR-103a was not recommended because its stability value exceeded the reference value. Thus, we combined stable genes and investigated the stability and the effect of gene normalization. The combination of miR-24, miR-103a, and let-7a was identified as one of the most stable sets of endogenous reference genes for normalization in FFPE lymph node. This study may provide a basis for miRNA expression analysis in FFPE lymph node tissue.

引用

页数：10

共 28 条

[11] The Heterochronic microRNA let-7 Inhibits Cell Motility by Regulating the Genes in the Actin Cytoskeleton Pathway in Breast Cancer [J].