Development of an Icariin-Loaded Bilosome-Melittin Formulation with Improved Anticancer Activity against Cancerous Pancreatic Cells

Pancreatic cancer currently represents a severe issue for the entire world. Therefore, much effort has been made to develop an effective treatment against it. Emerging evidence has shown that icariin, a flavonoid glycoside, is an effective anti-pancreatic cancer drug. Melittin, as a natural active biomolecule, has also shown to possess anticancer activities. In the present study, with the aim to increase its effectiveness against cancerous cells, icariin-loaded bilosome-melittin (ICA-BM) was developed. For the selection of an optimized ICA-BM, an experimental design was implemented, which provided an optimized formulation with a particle size equal to 158.4 nm. After estimation of the release pattern, the anti-pancreatic cancer efficacy of this new formulation was evaluated. The MTT assay was employed for the determination of half maximal inhibitory concentration (IC50), providing smaller IC50 for ICA-BM (2.79 ± 0.2 µM) compared to blank-BM and ICA-Raw (free drug) against PNAC1, a human pancreatic cancer cell line isolated from a pancreatic carcinoma of ductal cell origin. Additionally, cell cycle analysis for ICA-BM demonstrated cell arrest at the S-phase and pre-G1 phase, which indicated a pro-apoptotic behavior of the new developed formulation. The pro-apoptotic and anti-proliferative activity of the optimized ICA-BM against PNAC1 cells was also demonstrated through annexin V staining as well as estimation of caspase-3 and p53 protein levels. It can be concluded that the optimized ICA-BM formulation significantly improved the efficacy of icariin against cancerous pancreatic cells.

REVIEW OF POTENTIAL PLANTS IN INDONESIA AS AN ANTIDIABETIC WITH HYPOGLYCEMIC ACTIVITIES

Background: Diabetes mellitus which is characterized by hyperglycemia is a metabolic disease due to insulin action, defects in insulin secretion, or both. Various studies have shown that herbs that have a hypoglycemic effect can be used in the treatment of diabetes mellitus. Generally, herbs can slow down the complications of diabetes mellitus and improve metabolism. The ability of plants to restore the function of pancreatic tissue to increase insulin production or make it easier for insulin to process glucose is related to the effects of hypoglycemia. Objective: Comparing plants that have a hypoglycemic activity that can be used in the treatment of diabetes mellitus in Indonesia. Methods: studying literature from various databases, websites, national journals, and relevant international journals to identify plants that have hypoglycemic activity. Results: The mechanism A. sativum as a hypoglycemic involves different fiber viscosities; A. vera involved as a hypoglycemic by protecting pancreatic? cells; Asiatic acid in C. asiatica increasing glycolysis by restoring the activity of key enzymes; G. max inhibiting ?-amylase and the stigmasterol increasing the absorption of blood glucose and reducing insulin resistance; P. macrocarpa exert their antidiabetic action via ?-glucosidase modulation, an extra distinctive pancreatic mechanism; S. arvensis has the potential to inhibit ?-amylase enzymes. Conclusions: This review article has presented 6 detailed comparisons of plants that have a hypoglycemic activity that can be used in the treatment of diabetes mellitus in Indonesia and we believe can be useful for students, researchers, or practitioners. Keywords: Diabetes mellitus, hypoglycemia, Indonesian plants

KrasG12D induces changes in chromatin territories that differentially impact early nuclear reprogramming in pancreatic cells

Background Pancreatic ductal adenocarcinoma initiation is most frequently caused by Kras mutations. Results Here, we apply biological, biochemical, and network biology methods to validate GEMM-derived cell models using inducible KrasG12D expression. We describe the time-dependent, chromatin remodeling program that impacts function during early oncogenic signaling. We find that the KrasG12D-induced transcriptional response is dominated by downregulated expression concordant with layers of epigenetic events. More open chromatin characterizes the ATAC-seq profile associated with a smaller group of upregulated genes and epigenetic marks. RRBS demonstrates that promoter hypermethylation does not account for the silencing of the extensive gene promoter network. Moreover, ChIP-Seq reveals that heterochromatin reorganization plays little role in this early transcriptional program. Notably, both gene activation and silencing primarily depend on the marking of genes with a combination of H3K27ac, H3K4me3, and H3K36me3. Indeed, integrated modeling of all these datasets shows that KrasG12D regulates its transcriptional program primarily through unique super-enhancers and enhancers, and marking specific gene promoters and bodies. We also report chromatin remodeling across genomic areas that, although not contributing directly to cis-gene transcription, are likely important for KrasG12D functions. Conclusions In summary, we report a comprehensive, time-dependent, and coordinated early epigenomic program for KrasG12D in pancreatic cells, which is mechanistically relevant to understanding chromatin remodeling events underlying transcriptional outcomes needed for the function of this oncogene.

Anti-apoptotic effect of Buchholzia coriacea Engl. stem back extracts on AsPC-1 and mechanisms of action

Ethnopharmacological relevance Buchholzia coriacea Engl. is popularly called wonderful cola due to its wide ethnomedicinal use for the treatment of various ailments. We investigated the possible cytotoxic effect of its various fractions on human pancreatic cancer cell (AsPC-1) and also determined its mechanisms of action. Materials and methods The AsPC-1 cells were cultivated and separately treated with 5-fluorouracil (5-FU) or Buchholzia coriacea Engl. bark (BC) (ethanol, aqueous, chloroform or ethyl acetate extract) for 72 h. Cell viability, caspase 3 and mitochondrial membrane potential (ΔΨm) were determined in vitro after the treatment. Nitric oxide (NO) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals’ scavenging property, ferric reducing power and lipid peroxidation assays were also done to examine the antioxidant effect of BC in vitro. Results Various extracts of BC, especially at 2500 μg/ml and 5000 μg/ml, increased the AsPC-1 viability while 5-FU decreased it. The activity of caspase 3 was increased by 5-FU but reduced by all concentrations of various extracts of BC. Incubation of AsPC-1 with 5-FU showed the majority of cells having the monomeric form of JC-1 dye (bright green fluorescence), which indicated de-energized mitochondria. However, fluorescence photomicrograph of cells incubated with different concentrations (20, 40 and 100 μg/ml) of BC extracts (aqueous, ethanol, chloroform and ethyl acetate) showed strong JC-1 aggregation (yellow), which indicated mitochondria with intact membrane potentials. BC extracts also scavenged NO and DPPH radicals, inhibited lipid peroxidation and increased ferric reduction, though not as much as ascorbic acid. Conclusion This study suggests that BC elicits anti-apoptotic activity in AsPC-1 by increasing cell viability, decreasing caspase 3 activity, stabilizing the ∆Ψm, and scavenging free radicals. Even though BC is used ethnomedicinally as anti-cancer agent, our findings in the present study suggest that it has pro-cancer potential in-vitro, especially on pancreatic cells. Its anti-apoptotic activity in AsPC-1 could be of clinical significance, especially to counteract the effect of apoptotic agents on pancreatic cells.

Gene Signatures of NEUROGENIN3+ Endocrine Progenitor Cells in the Human Pancreas

NEUROGENIN3+ (NEUROG3+) cells are considered to be pancreatic endocrine progenitors. Our current knowledge on the molecular program of NEUROG3+ cells in humans is largely extrapolated from studies in mice. We hypothesized that single-cell RNA-seq enables in-depth exploration of the rare NEUROG3+ cells directly in humans. We aligned four large single-cell RNA-seq datasets from postnatal human pancreas. Our integrated analysis revealed 10 NEUROG3+ epithelial cells from a total of 11,174 pancreatic cells. Noticeably, human NEUROG3+ cells clustered with mature pancreatic cells and epsilon cells displayed the highest frequency of NEUROG3 positivity. We confirmed the co-expression of NEUROG3 with endocrine markers and the high percentage of NEUROG3+ cells among epsilon cells at the protein level based on immunostaining on pancreatic tissue sections. We further identified unique genetic signatures of the NEUROG3+ cells. Regulatory network inference revealed novel transcription factors including Prospero homeobox protein 1 (PROX1) may act jointly with NEUROG3. As NEUROG3 plays a central role in endocrine differentiation, knowledge gained from our study will accelerate the development of beta cell regeneration therapies to treat diabetes.

Dimethylaminomicheliolide (DMAMCL) Inhibits Cell Proliferation and Increases Apoptosis and Efficacy of Gemcitabine via Annexin A2 in Pancreatic Adenocarcinoma

Background: Pancreatic adenocarcinoma is one of the highest malignant tumors in digestive tract with extremely poor survival rate. Dimethylaminomicheliolide (DMAMCL) is a clinical developing anti-cancer agent, however, little is known regarding its effects in pancreatic cancer, and the mechanisms of DMAMCL are still not fully understood.Methods: This study evaluated DMAMCL on three pancreatic cancer cell lines by cell viability assay, colony formation assay, and apoptosis assay. To identify the direct binding target of DMAMCL in pancreatic cells, a chemical proteomics approach, molecular docking and site-directed mutagenesis were performed.Results: DMAMCL inhibits proliferation and promotes apoptosis of pancreatic cancer cells. Interestingly, using a chemical proteomics approach, we identify ANXA2 as a direct binding target of DMAMCL in pancreatic cells. Molecular docking and site-directed mutagenesis confirm that Cys132 (C132) of ANXA2 is the binding site for DMAMCL. The knockdown of ANXA2 largely decreases the inhibition activity of DMAMCL, indicating that the effect of DMAMCL is mainly mediated by ANXA2 in pancreatic cancer. In addition, the combination regimen of gemcitabine and DMAMCL exhibits synergistic effect on pancreatic cancer cell lines at both proliferation and pro-apoptosis level. Conclusions: Thus, our findings elucidate the mechanisms of DMAMCL and may provide a potential strategy to enhancing the efficacy of gemcitabine in pancreatic adenocarcinoma.