PMID- 36627346
OWN - NLM
STAT- MEDLINE
DCOM- 20230112
LR  - 20230228
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 13
IP  - 1
DP  - 2023 Jan 10
TI  - Identification of stable reference genes in peripheral blood mononuclear cells 
      from type 2 diabetes mellitus patients.
PG  - 486
LID - 10.1038/s41598-023-27460-3 [doi]
LID - 486
AB  - Reference genes are obligatory for accurate normalization of mRNA transcript 
      levels across samples and experimental conditions in Real Time-polymerase chain 
      reaction (qRT-PCR) based quantitative gene expression assays. Selection of stably 
      expressed reference genes is therefore crucial for ensuring reproducibility of 
      such assays. However, there is a complete dearth of data on stability of commonly 
      used reference genes in Peripheral Blood Mononuclear Cells (PBMCs) from Type 2 
      diabetes mellitus (T2DM) patients. We have evaluated the gene expression 
      stability of 4 widely used reference genes (Beta-actin, ACTB; Peptidylprolyl 
      Isomerase B, PPIB; Tyrosine 3 Monooxygenase/Tryptophan 5-Monooxygenase Activation 
      Protein Zeta, YWHAZ; and Glyceraldehyde-3-Phosphate Dehydrogenase, GAPDH); in 
      PBMCs from 39 T2DM patients and 47 normoglycemic (NGT) subjects. ACTB and YWHAZ 
      were found to be the most stable genes in PBMCs from T2DM patients and therefore, 
      can be recommended as suitable reference genes in similar contexts. GAPDH and 
      PPIB expressions were not stable in PBMCs from T2DM patients. On using ACTB and 
      YWHAZ as reference genes for measuring relative expression of GAPDH and PPIB in 
      these subjects, relative GAPDH expression was found to be significantly lower in 
      female T2DM patients, compared to female NGT subjects [GAPDH relative 
      normalization unit (RNU): female T2DM (n = 19), median (Q1, Q3): 9.0 (8.1, 9.9); 
      female NGT (n = 18): median (Q1, Q3): 10.1 (9.1, 11.0); P = 0.034]. Dysregulation 
      of GAPDH in PBMCs from female T2DM patients could be associated with sex-specific 
      differences in pathogenesis and outcomes of T2DM.
CI  - (c) 2023. The Author(s).
FAU - Hazarika, Ankita
AU  - Hazarika A
AD  - Division of Nutrition, St. John's Research Institute, St. John's National Academy 
      of Health Sciences, Sarjapur Road, Bangalore, India.
FAU - Nongkhlaw, Bajanai
AU  - Nongkhlaw B
AD  - Division of Nutrition, St. John's Research Institute, St. John's National Academy 
      of Health Sciences, Sarjapur Road, Bangalore, India.
AD  - Department of Pathology, North Eastern Indira Gandhi Regional Institute of Health 
      and Medical Sciences, Shillong, Meghalaya, India.
FAU - Mukhopadhyay, Arpita
AU  - Mukhopadhyay A
AUID- ORCID: 0000-0001-8260-5385
AD  - Division of Nutrition, St. John's Research Institute, St. John's National Academy 
      of Health Sciences, Sarjapur Road, Bangalore, India. arpitam@sjri.res.in.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230110
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Male
MH  - Humans
MH  - Female
MH  - *Leukocytes, Mononuclear
MH  - Reproducibility of Results
MH  - *Diabetes Mellitus, Type 2/genetics
MH  - Real-Time Polymerase Chain Reaction
MH  - RNA, Messenger/genetics
MH  - Reference Standards
MH  - Gene Expression Profiling
PMC - PMC9831022
COIS- The authors declare no competing interests.
EDAT- 2023/01/11 06:00
MHDA- 2023/01/13 06:00
PMCR- 2023/01/10
CRDT- 2023/01/10 23:18
PHST- 2022/06/16 00:00 [received]
PHST- 2023/01/02 00:00 [accepted]
PHST- 2023/01/10 23:18 [entrez]
PHST- 2023/01/11 06:00 [pubmed]
PHST- 2023/01/13 06:00 [medline]
PHST- 2023/01/10 00:00 [pmc-release]
AID - 10.1038/s41598-023-27460-3 [pii]
AID - 27460 [pii]
AID - 10.1038/s41598-023-27460-3 [doi]
PST - epublish
SO  - Sci Rep. 2023 Jan 10;13(1):486. doi: 10.1038/s41598-023-27460-3.