Selection of Endogenous Control Reference Genes for Studies on Type 1 or Type 2 Endometrial Cancer

被引：22

作者：

Ayakannu, Thangesweran ^{[1
,2
,3
]}

Taylor, Anthony H. ^{[1
,4
]}

Konje, Justin C. ^{[1
,5
]}

机构：

[1] Univ Liverpool, Fac Hlth & Life Sci, Thompson Yates Brownlow Hill, Liverpool L69 3GB, Merseyside, England

[2] Univ Leicester, Dept Canc Studies & Mol Med, Reprod Sci Sect, Leicester, Leics, England

[3] Liverpool Womens NHS Fdn Trust, Liverpool Womens Hosp, Gynaecol Oncol Canc Ctr, Liverpool, Merseyside, England

[4] Univ Leicester, Dept Mol & Cell Biol, Leicester, Leics, England

[5] HMC, Dept Obstet & Gynaecol, Womens Wellness & Res Ctr, Sidra Med, Doha, Qatar

来源：

SCIENTIFIC REPORTS | 2020年 / 10卷 / 01期

关键词：

REAL-TIME PCR; RT-PCR; EXPRESSION; TRANSCRIPTION; RECEPTOR; CARCINOMA; QUANTIFICATION; NORMALIZATION; VALIDATION; BIOMARKERS;

D O I：

10.1038/s41598-020-64663-4

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：

A panel of 32 candidate reference genes was used to identify the most stable genes for gene normalisation in quantitative RT-PCR studies using endometrial biopsies obtained from women with endometrial cancer (type 1 or type 2) and without cancer (controls). RNA from the biopsies was isolated, examined for purity and quality, and then reverse transcribed into cDNA before being subjected to real-time qRT-PCR analysis in triplicate within the TaqMan gene Expression Assay kit. The most 'stable' endogenous control genes were then identified using the geNorm qbase+2 and NormFinder software packages. PSMC4, PUM1 and IPO8 were identified as the best reference genes combination for type 1 endometrial cancer (grades 1, 2 and 3), whereas for type 2 endometrial cancer (serous and carcinosarcoma), UBC, MRPL19, PGK1 and PPIA were the best reference genes combination. We conclude that the use of these normaliser combinations should provide accurate interpretation of gene expression at the transcript level in endometrial cancer studies especially for types 1 and 2 cancers.

引用

页数：10

共 41 条

[1] Impact of reference gene selection for type 2 cannabinoid receptor gene expression studies in human spermatozoa [J].