P Kochhar1, P Dwarkanath1, G Ravikumar2, A Thomas3, J Crasta2, T Thomas4, A V Kurpad1, A Mukhopadhyay5. 1. Division of Nutrition, St. John's Research Institute, A recognized research centre of University of Mysore, Bangalore, Karnataka, India. 2. Department of Pathology, St John's Medical College Hospital, Bangalore, Karnataka, India. 3. Department of Obstetrics and Gynaecology, St John's Medical College Hospital, Bangalore, Karnataka, India. 4. Department of Biostatistics, St. John's Medical College Hospital, Bangalore, Karnataka, India. 5. Division of Nutrition, St. John's Research Institute, A recognized research centre of University of Mysore, Bangalore, Karnataka, India. arpitam@sjri.res.in.
Abstract
BACKGROUND/ OBJECTIVES: The current study aimed to identify suitable reference miRNA for placental miRNA expression analysis in a set of well-characterized and fetal-sex balanced small- (SGA) and appropriate- (AGA) for gestational age full-term singleton pregnancies. SUBJECTS/ METHODS: In this retrospective study, placental samples (n = 106) from 35 SGA (19 male and 16 female) and 71 AGA (30 male and 41 female) full-term singleton pregnancies were utilized. Placental transcript abundance of three widely used reference miRNAs [miR-16-5p and Small nucleolar RNAs (snoRNAs) RNU44 and RNU48] were assessed by real-time quantitative PCR. Raw cycle threshold (Ct) analysis and RefFinder tool analysis were conducted for evaluating stability of expression of these miRNAs. RESULTS: Raw Ct values of miR-16-5p were similar between SGA and AGA births (P = 0.140) and between male and female births within SGA (P = 0.159) and AGA (P = 0.060) births while that of RNU44 and RNU48 were higher in SGA births (P = 0.008 and 0.006 respectively) and in male births within the SGA group (P = 0.005) for RNU44 and in female births within the AGA group (P = 0.048) for RNU48. Across all 106 samples tested using the RefFinder tool, miR-16-5p and RNU44 were equally stable reference miRNAs. CONCLUSION: We recommend miR-16-5p and RNU44 as suitable reference miRNAs for placental samples from settings similar to our study.
BACKGROUND/ OBJECTIVES: The current study aimed to identify suitable reference miRNA for placental miRNA expression analysis in a set of well-characterized and fetal-sex balanced small- (SGA) and appropriate- (AGA) for gestational age full-term singleton pregnancies. SUBJECTS/ METHODS: In this retrospective study, placental samples (n = 106) from 35 SGA (19 male and 16 female) and 71 AGA (30 male and 41 female) full-term singleton pregnancies were utilized. Placental transcript abundance of three widely used reference miRNAs [miR-16-5p and Small nucleolar RNAs (snoRNAs) RNU44 and RNU48] were assessed by real-time quantitative PCR. Raw cycle threshold (Ct) analysis and RefFinder tool analysis were conducted for evaluating stability of expression of these miRNAs. RESULTS: Raw Ct values of miR-16-5p were similar between SGA and AGA births (P = 0.140) and between male and female births within SGA (P = 0.159) and AGA (P = 0.060) births while that of RNU44 and RNU48 were higher in SGA births (P = 0.008 and 0.006 respectively) and in male births within the SGA group (P = 0.005) for RNU44 and in female births within the AGA group (P = 0.048) for RNU48. Across all 106 samples tested using the RefFinder tool, miR-16-5p and RNU44 were equally stable reference miRNAs. CONCLUSION: We recommend miR-16-5p and RNU44 as suitable reference miRNAs for placental samples from settings similar to our study.
Authors: Yu Wang; Eugenie R Lumbers; Anya L Arthurs; Celine Corbisier de Meaultsart; Andrea Mathe; Kelly A Avery-Kiejda; Claire T Roberts; Fiona Broughton Pipkin; Francine Z Marques; Brian J Morris; Kirsty G Pringle Journal: Mol Hum Reprod Date: 2018-09-01 Impact factor: 4.025