AUM302 inhibits the cell signaling pathways regulated by PIM kinases and PI3K/mTOR pathway.

BxPC-3 (A), Capan-2 (B), MIA PaCa-2 (C), PANC-1 (D), and Hs766T (E) cells were treated with DMSO (vehicle) or TP-3654 (10 and 100 nM) or AUM302 (10 and 100 nM) for 24 hours.</p

AUM302 changes the cell cycle profile of Hs766T pancreatic cancer cell line.

Cells were treated with DMSO or TP-3654 (100 nM) or AUM302 (100 nM) for 24 (A), 48 (B), and 72 hours (C). Cells were stained with propidium iodide and analyzed by FACS analysis. Data are represented as mean ±SD, N = 3, *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 calculated with two-way ANOVA.</p

AUM302 and TP-3654 decrease the cell viability of MIA PaCa-2 gemcitabine-resistant (GemR) cell line.

MIA PaCa-2 GemR cell line was treated with 10 nM, 100 nM, or 1 μM of TP-3654 (A, B, & C) or AUM302 (D, E, & F) twenty-four hours after seeding. Cells were treated with test compounds for 72 hours and cell viability was measured using Cell Titer-Glo. Data represents mean ±SD (N = 6 and N = 4). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 calculated with two-way ANOVA.</p

AUM302 inhibits the proliferation of MIA PaCa-2 gemcitabine-resistant (GemR) cells and activity of multiple signaling pathways.

(A) MIA PaCa-2 GemR cells were treated with 10 nM, 100 nM, and 1 μM of TP-3654 or AUM302. Cell count was determined 24, 48, and 72 hours after treatment using a cell counter. The measurement of the control (cells with DMSO) was defined as 100%. Data represent mean ±SD (N = 9). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 calculated with two-way ANOVA. (B—D) MIA PaCa-2 GemR cells were treated with DMSO (vehicle) or TP-3654 or AUM302 (10 nM, 100 nM, and 1 μM) for 24 (B), 48 (C), and 72 (D) hours and analyzed with western blot.</p

AUM302 inhibits the proliferation of multiple pancreatic cancer cell lines.

The following pancreatic cancer cell lines, BxPC-3 (A), Capan-2 (B), MIA PaCa-2 (C), PANC-1 (D), and Hs766T (E), were treated with DMSO or TP-3654 (10 nM and 100 nM), or AUM302 (10 nM and 100 nM). Cell count was determined 24, 48, and 72 hours after treatment using a cell counter. The measurement of the control (cells with DMSO) was defined as 100%. Data represent mean ±SD (N = 6). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 calculated with two-way ANOVA.</p

Densitometry analysis of western blots results of proteins regulated by PIM and PI3K/mTOR pathway in MIA PaCa-2 GemR cells treated with 10 nM, 100 nM, and 1 μM of TP-3654 or AUM302 for 24 hours.

Each experiment was performed in triplicate and the results are shown as mean ±SD (N = 3). Densitometry analysis was performed using FIJI software [61]. Statistical analysis was performed using the Student’s test followed by an analysis of the normal distribution (Tukey’s test). *p (PDF)</p

Ribosomal proteins are downregulated in AA CRC vs CA CRC.

<p>Ribosomal proteins are downregulated in AA CRC vs CA CRC.</p

Differently methylated miRNAs in AA and CA CRC specimens, ranked by statistical significance.

<p>Differently methylated miRNAs in AA and CA CRC specimens, ranked by statistical significance.</p

Aberrant DNA Methylation: Implications in Racial Health Disparity

<div><p>Background</p><p>Incidence and mortality rates of colorectal carcinoma (CRC) are higher in African Americans (AAs) than in Caucasian Americans (CAs). Deficient micronutrient intake due to dietary restrictions in racial/ethnic populations can alter genetic and molecular profiles leading to dysregulated methylation patterns and the inheritance of somatic to germline mutations.</p><p>Materials and Methods</p><p>Total DNA and RNA samples of paired tumor and adjacent normal colon tissues were prepared from AA and CA CRC specimens. Reduced Representation Bisulfite Sequencing (RRBS) and RNA sequencing were employed to evaluate total genome methylation of 5’-regulatory regions and dysregulation of gene expression, respectively. Robust analysis was conducted using a trimming-and-retrieving scheme for RRBS library mapping in conjunction with the BStool toolkit.</p><p>Results</p><p>DNA from the tumor of AA CRC patients, compared to adjacent normal tissues, contained 1,588 hypermethylated and 100 hypomethylated differentially methylated regions (DMRs). Whereas, 109 hypermethylated and 4 hypomethylated DMRs were observed in DNA from the tumor of CA CRC patients; representing a 14.6-fold and 25-fold change, respectively. Specifically; CHL1, 4 anti-inflammatory genes (i.e., NELL1, GDF1, ARHGEF4, and ITGA4), and 7 miRNAs (of which miR-9-3p and miR-124-3p have been implicated in CRC) were hypermethylated in DNA samples from AA patients with CRC. From the same sample set, RNAseq analysis revealed 108 downregulated genes (including 14 ribosomal proteins) and 34 upregulated genes (including POLR2B and CYP1B1 [targets of miR-124-3p]) in AA patients with CRC versus CA patients.</p><p>Conclusion</p><p>DNA methylation profile and/or products of its downstream targets could serve as biomarker(s) addressing racial health disparity.</p></div

Annotated list of differentially downregulated genes in AA CRC compared to CA CRC, ranked by statistical significance.

<p>Annotated list of differentially downregulated genes in AA CRC compared to CA CRC, ranked by statistical significance.</p