1.Metabolic Interplay between Tumour Cells and Cancer-Associated Fibroblasts (CAFs) under Hypoxia versus Normoxia
Septelia Inawati WANANDI ; Sri Suciati NINGSIH ; Hijrah ASIKIN ; Rendy HOSEA ; Gladies Mercya Grameinie Neolaka
Malaysian Journal of Medical Sciences 2018;25(3):7-16
The growth of tumour cells is closely related to cancer-associated fibroblasts (CAFs)present within their microenvironment. CAFs, the most abundant cells in tumour stroma, secretegrowth factors that play pivotal roles in tumour cell proliferation, metabolism, angiogenesis andmetastasis. Tumour cells adapt to rapid environmental changes from normoxia to hypoxia throughmetabolic interplay with CAFs. In this mini review, we discuss the role of lactate dehydrogenases(LDHs) and monocarboxylate transporters (MCTs) on the metabolic interplay between tumourcells and CAFs under hypoxia compared to normoxia. The LDHs catalyse the interchange oflactate and pyruvate, whereas MCTs facilitate the influx and efflux of monocarboxylates, especiallylactate and pyruvate. To sum up, tumour cells switch their metabolic state between glycolysis andoxidative phosphorylation through metabolic interplay with CAFs, which exhibit the Warburgeffect under hypoxia and reverse Warburg effect under normoxia.
2.Gene Expression of Molecules Regulating Apoptotic Pathways in Glioblastoma Multiforme Treated with Umbilical Cord Stem Cell Conditioned Medium
Novi Silvia Hardiany ; Edward Christopher Yo ; Eko Ngadiono ; Septelia Inawati Wanandi
Malaysian Journal of Medical Sciences 2019;26(6):35-45
Background: Glioblastoma multiforme (GBM) is the most malignant primary brain
tumour and there is no definite cure. It has been suggested that there are significant interactions
among mesenchymal stem cells (MSCs), their released factors and tumour cells that ultimately
determine GBM’s growth pattern. This study aims to analyse the expression of molecules involved
in GBM cell apoptotic pathways following treatment with the MSC secretome.
Methods: A conditioned medium of umbilical cord-derived MSCs (UCMSC-CM) was
generated by culturing the cells on serum-free αMEM for 24 h. Following this, human GBM T98G
cells were treated with UCMSC-CM for 24 h. Quantitative reverse transcriptase-polymerase chain
reaction (qRT-PCR) was then performed to measure the mRNA expression of survivin, caspase-9,
TNF-related apoptosis-inducing ligand (TRAIL), DR4 and DcR1.
Results: mRNA expression of caspase-9 in CM-treated T98G cells increased 1.6-fold
(P = 0.017), whereas mRNA expression of survivin increased 3.5-fold (P = 0.002). On the other
hand, TRAIL protein expression was upregulated (1.2-fold), whereas mRNA expression was
downregulated (0.4-fold), in CM-treated cells. Moreover, there was an increase in the mRNA
expression of both DR4 (3.5-fold) and DcR1 (1,368.5-fold) in CM-treated cells.
Conclusion: The UCMSC-CM was able to regulate the expression of molecules involved
in GBM cell apoptotic pathways. However, the expression of anti-apoptotic molecules was more
upregulated than that of pro-apoptotic molecules.
3.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.