Gene expression profiling of acute myeloid leukemia samples from adult patients with AML-M1 and -M2 through boutique microarrays, real-time PCR and droplet digital PCR

Abstract

Acute myeloid leukemia (AML) is the most common and severe form of acute leukemia diagnosed in adults. Owing to its heterogeneity, AML is divided into classes associated with different treatment outcomes and specific gene expression profiles. Based on previous studies on AML, in this study, we designed and generated an AML-array containing 900 oligonucleotide probes complementary to human genes implicated in hematopoietic cell differentiation and maturation, proliferation, apoptosis and leukemic transformation. The AML-array was used to hybridize 118 samples from 33 patients with AML of the M1 and M2 subtypes of the French-American-British (FAB) classification and 15 healthy volunteers (HV). Rigorous analysis of the microarray data revealed that 83 genes were differentially expressed between the patients with AML and the HV, including genes not yet discussed in the context of AML pathogenesis. The most overexpressed genes in AML were STMN1, KITLG, CDK6, MCM5, KRAS, CEBPA, MYC, ANGPT1, SRGN, RPLP0, ENO1 and SET, whereas the most underexpressed genes were IFITM1, LTB, FCN1, BIRC3, LYZ, ADD3, S100A9, FCER1G, PTRPE, CD74 and TMSB4X. The overexpression of the CPA3 gene was specific for AML with mutated NPM1 and FLT3. Although the microarray-based method was insufficient to differentiate between any other AML subgroups, quantitative PCR approaches enabled us to identify 3 genes (ANXA3, S100A9 and WT1) whose expression can be used to discriminate between the 2 studied AML FAB subtypes. The expression levels of the ANXA3 and S100A9 genes were increased, whereas those of WT1 were decreased in the AML-M2 compared to the AML-M1 group. We also examined the association between the STMN1, CAT and ABL1 genes, and the FLT3 and NPM1 mutation status. FLT3+/NPM1− AML was associated with the highest expression of STMN1, and ABL1 was upregulated in FLT3+ AML and CAT in FLT3− AML, irrespectively of the NPM1 mutation status. Moreover, our results indicated that CAT and WT1 gene expression levels correlated with the response to therapy. CAT expression was highest in patients who remained longer under complete remission, whereas WT1 expression increased with treatment resistance. On the whole, this study demonstrates that the AML-array can potentially serve as a first-line screening tool, and may be helpful for the diagnosis of AML, whereas the differentiation between AML subgroups can be more successfully performed with PCR-based analysis of a few marker genes.

Enhanced expression of FCER1G predicts positive prognosis in multiple myeloma

“…Associated diseases included Bleeding Disorder, Platelet-Type, 11(BDPLT11) and Mitochondrial Complex I Deficiency. FCER1G also engaged in many immune responses and played a tumor-promoting role in many kinds of tumors, such as meningioma, Clear cell renal cell carcinoma (ccRCC), childhood leukemia and Acute Myeloid Leukemia(AML) [12,14,15,30]. It was also reported that the demethylation of FCER1G was induced by IL15 in the NKp30+CD8+ T cell population exhibiting high natural killer-like antitumor potential [31].…”

Unlikely role of glycolytic enzyme ��-enolase in cancer metastasis and its potential as a prognostic biomarker

“…The Warburg effect has been hypothesised to be an adaptation mechanism in cancer cells to support the biosynthetic requirements of rapid proliferation. Alpha-enolase expression has been shown to be altered at the mRNA and/or protein level in a range of tumours [ Table 1], and generally upregulated in most, including acute myeloid leukaemia (AML), glioma, melanoma, lymphoma, and colorectal, endometrial, gastric, head and neck, liver, ovarian and pancreatic cancer [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22] .…”

“…Microarray Increased [4] Breast cancer Breast cancer tissue with adjacent matched normal tissue (n = 24)…”

“…Increased glycolysis is considered a hallmark of cancer [2] and investigating glycolytic enzymes may yield new therapeutic approaches for cancer treatment. Enolases are key glycolytic enzymes [3] , and increased expression of one isoform, a-enolase, has been identified in several cancer types [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22] . This review provides an overview of the expression of a-enolase and key functions it controls in cancer cells, with a focus on the potential role of a-enolase as a cancer prognostic biomarker or therapeutic target.…”

The implication of lncRNA expression pattern and potential function of lncRNA RP4‐576H24.2 in acute myeloid leukemia

“…As for the possible explanations to our results, it might include: 1) lncRNA RP4-576H24.2 may promote development and progression of AML through regulating multiple downstream genes as displayed in our RNA sequencing results, in which several genes regulated by lncRNA RP4-576H24.2 are reported to participate in the pathogenesis of leukemia, such as PTPRJ, FCN1, KLF4, G0S2, and LILRB2. [28][29][30][31][32] Therefore, ln-cRNA RP4-576H24.2 might promote the development or progression of AML by mediating the leukemia pathology-related genes, such as PTPRJ, FCN1, KLF4, G0S2, and LILRB2; 2) our further cell experiments disclosed that lncRNA RP4-576H24.2 promoted proliferation while inhibited apoptosis of two AML cell lines, indicating that lncRNA RP4-576H24.2 might aggravate the progression of AML through regulating AML cell functions.…”