摘要：Multiple myeloma (MM) is a cancer of malignant plasma cells in the bone marrow
which might lead to the development of one or more clinical manifestations such as
bone destruction, anemia, renal insufficiency etc. Gene expression analysis using
quantitative real-time PCR (qRT-PCR) is imperative to understand the
developmental mechanisms of MM. Housekeeping genes (HKGs) are commonly
used as endogenous controls to normalize quantitative real-time PCR (qRT-PCR)
data for gene expression analysis. However, recent studies argue that the expression
of HKG genes may vary under certain experimental condition. In addition, no
studies have been found on the expression analysis of HKGs by qRT-PCR in MM.
Therefore, the present study was designed to validate reference genes for qRT-PCR
normalization through observing the effects of hypoxia, serum stimulation and Myc
inhibition on the expression of five HKGs (18S, ACTB, B2M, GAPDH, and TBP) in
three different myeloma cell lines (ANBL-6, IH-1 and INA-6). Four different
approaches (Best Keeper, ∆Ct approach, GeNorm, and NormFinder) followed by
comprehensive methods were used for the evaluation and selection of reference
gene. Most stable expression of 18S in hypoxic and serum stimulation experiment
while constant expression of B2M in Myc inhibition made them an excellent
combination to normalize qRT-PCR data in gene expression analysis in MM.
