1.Profiling of Gene Expression Associated with Stemness and Aggressiveness of ALDH1A1- Expressing Human Breast Cancer Cells
Septelia Inawati Wanandi ; Resda Akhra Syahrani ; Sekar Arumsari ; Gita Wideani ; Novi Silvia Hardiany
Malaysian Journal of Medical Sciences 2019;26(5):38-52
Background: It has been widely reported that breast cancer aggressiveness may be driven
by breast cancer stem cells (BCSCs). BCSCs display stemness properties that include self-renewal,
tumourigenicity and pluripotency. The regulation of gene expression may have important roles in
BCSC stemness and aggressiveness. Thus, the aim of this study was to examine the stemness and
aggressiveness gene expression profile of BCSCs compared to MCF-7 and MDA-MB-231 breast
cancer cells.
Methods: Human ALDH1+ BCSCs were grown in serum-free Dulbecco’s Modified Eagle
Medium (DMEM)/F12, while MCF-7 and MDA-MB-231 were cultured in DMEM supplemented with
10% foetal bovine serum under standard conditions. Total RNA was extracted using the Tripure
Isolation Reagent. The relative mRNA expressions of OCT4, ALDH1A1 and CD44 associated with
stemness as well as TGF-β1, TβR1, ERα1 and MnSOD associated with aggressiveness in BCSCs and
MCF-7 cells were determined using the quantitative real-time PCR (qRT-PCR).
Results: The mRNA expressions of OCT4 (5.19-fold ± 0.338; P = 0.001), ALDH1A1 (3.67-
fold ± 0.523; P = 0.006), CD44 (2.65-fold ± 0.307; P = 0.006), TGF-β1 (22.89-fold ± 6.840; P =
0.015), TβR1 (3.74-fold ± 1.446; P = 0.045) and MnSOD (4.6-fold ± 1.096; P = 0.014) were higher
in BCSCs than in MCF-7 but were almost similar to MDA-MB-231 cells. In contrast, the ERα1
expression of BCSCs (0.97-fold ± 0.080; P = 0.392) was similar to MCF-7 cells, indicating that
BSCSs are oestrogen-dependent breast cancer cells.
Conclusion: The oestrogen-dependent BCSCs express stemness and aggressiveness genes
at a higher level compared to oestrogen-dependent MCF-7 but are almost similar to oestrogenindependent
MDA-MB-231 cells.