Lytic Bacteriophage EFA1 Modulates HCT116 Colon Cancer Cell Growth and Upregulates ROS Production in an Enterococcus faecalis Co-culture System

Enterococcus faecalis is an opportunistic pathogen in the gut microbiota that’s associated with a range of difficult to treat nosocomial infections. It is also known to be associated with some colorectal cancers. Its resistance to a range of antibiotics and capacity to form biofilms increase its virulence. Unlike antibiotics, bacteriophages are capable of disrupting biofilms which are key in the pathogenesis of diseases such as UTIs and some cancers. In this study, bacteriophage EFA1, lytic against E. faecalis, was isolated and its genome fully sequenced and analyzed in silico. Electron microscopy images revealed EFA1 to be a Siphovirus. The bacteriophage was functionally assessed and shown to disrupt E. faecalis biofilms as well as modulate the growth stimulatory effects of E. faecalis in a HCT116 colon cancer cell co-culture system, possibly via the effects of ROS. The potential exists for further testing of bacteriophage EFA1 in these systems as well as in vivo models.

Investigation of PTGS2, MAGE-A3, CALR, KRT19 and TMPRSS4 expressions in HCT116 colon cancer and PC3 prostate cancer cell lines

Cancer is a disease arising from DNA alterations that dysregulate gene structure and function. These deregulated genes can also play a role in tumor invasion and metastasis or resistance to treatment. In this study, we determined the gene expression during transcription of PTGS2 (Prostaglandin-endoperoxide synthase 2), MAGE-A3 (Melanoma-associated antigen 3), CALR (Calreticulin), KRT19 (Cytokeratin 19), and TMPRSS4 (Transmembrane protease, serine 4) in HCT116 colon cancer cell line and PC3 prostate cancer cell line. After RNA isolation and cDNA conversion, DNA amplification was performed with Real-Time PCR. We determined the altered transcriptional expression level of those genes. In HCT116 colon cancer cell line, expression of the TMPRSS4 gene, MAGEA3 gene and KRT19 gene was found as increased and expression of the CALR gene and the PTGS2 gene was found as decreased. Especially a 93.70-fold increase in expression of the KRT19 gene was found in HCT116 colon cancer cell line. In PC3 prostate cancer cell lines, TMPRS4 gene expression and MAGEA3 gene expression were found as increased. But there was 50 fold decrease in PTGS2 gene expression.

Anticancer Activity of Lindernia crustacea (L.) F. Muell. var. Crustacean on Human HCT116 Colon Cancer Cell via Cellular Lipid and β-sheet Protein Accumulation

Lindernia crustacea (L.) F. Muell. var. crustacean or “Ya Kap Hoi: YKH”, which is an edible vegetable, is commonly seen in Thailand. This study investigated its anticancer properties with high antioxidant activity by reducing power activity, excluding alkylation activity. It explored YKH extract induction of anticancer activity through biomolecular changes in the HCT116 human colon cancer cell line. The ethanolic extract stock solution and water extract stock solution were prepared. NR assay was used for cancer cell cytotoxic testing and Fourier transform infrared (FTIR) microspectroscopy was used for biomolecular changes study on lipid, protein and nucleic acid/DNA. The apoptosis induction by the extracts was detected by using Annexin V-FITC and DAPI staining. The compounds in the YKH ethanolic extract was performed by using GC-MS analysis. As a result, YKH ethanolic extract caused HCT116 colon cancer cell death in the dose dependent manner after 24 h exposure, and the 50 % cell death (IC50 concentration) was 195.4 ± 12 μg/mL. Cellular biochemical changes observed from FTIR data showed that YKH ethanolic extract treated HCT116 colon cancer cell. There is an increase in lipid content and a reduction in intensity of nucleic acid/DNA, a-helix protein structure at 1,656 cm-1 was reduced and peak position of b-sheet structure (1,637 cm-1) was shifted to lower frequency. From the analysis results, YKH ethanolic extract seems to exert anti-colon cancer effect by changing cellular biomolecular structure of lipids and β-sheet protein accumulation, supporting apoptotic induction. The compounds in the YKH ethanolic extract mainly yielded fatty acids,which may be useful as potential compounds.

Structural Modifications of Nature-Inspired Indoloquinolines: A Mini Review of Their Potential Antiproliferative Activity

Cryptolepine, neocryptolepine and isocryptolepine are naturally occurring indoloquinoline alkaloids with various spectrum of biological properties. Structural modification is an extremely effective means to improve their bioactivities. This review enumerates several neocryptolepine and isocryptolepine analogues with potent antiproliferative activity against MV4-11 (leukemia), A549 (lung cancer), HCT116 (colon cancer) cell lines in vitro. Its activity towards normal mouse fibroblasts BALB/3T3 was also evaluated. Furthermore, structure activity relationships (SAR) are briefly discussed. The anticancer screening of neocryptolepine derivatives was performed in order to determine their cytotoxic and growth inhibitory activities across the JFCR39 cancer cell line panel.

KO of 5-InsP7kinase activity transforms the HCT116 colon cancer cell line into a hypermetabolic, growth-inhibited phenotype

The inositol pyrophosphates 5-InsP7(diphosphoinositol pentakisphosphate) and 1,5-InsP8(bis-diphosphoinositol tetrakisphosphate) are highly energetic cellular signals interconverted by the diphosphoinositol pentakisphosphate kinases (PPIP5Ks). Here, we used CRISPR to KO PPIP5Ks in the HCT116 colon cancer cell line. This procedure eliminates 1,5-InsP8and raises 5-InsP7levels threefold. Expression of p53 and p21 was up-regulated; proliferation and G1/S cell-cycle transition slowed. Thus, PPIP5Ks are potential targets for tumor therapy. Deletion of the PPIP5Ks elevated [ATP] by 35%; both [ATP] and [5-InsP7] were restored to WT levels by overexpression of PPIP5K1, and a kinase-compromised PPIP5K1 mutant had no effect. This covariance of [ATP] with [5-InsP7] provides direct support for an energy-sensing attribute (i.e., 1 mMKmfor ATP) of the 5-InsP7–generating inositol hexakisphosphate kinases (IP6Ks). We consolidate this conclusion by showing that 5-InsP7levels are elevated on direct delivery of ATP into HCT116 cells using liposomes. Elevated [ATP] inPPIP5K−/−HCT116 cells is underpinned by increased mitochondrial oxidative phosphorylation and enhanced glycolysis. To distinguish between 1,5-InsP8and 5-InsP7as drivers of the hypermetabolic and p53-elevated phenotypes, we usedIP6K2RNAi and the pan-IP6K inhibitor,N2-(m-trifluorobenzyl),N6-(p-nitrobenzyl) purine (TNP), to return 5-InsP7levels inPPIP5K−/−cells to those of WT cells without rescuing 1,5-InsP8levels. Attenuation of IP6K restored p53 expression but did not affect the hypermetabolic phenotype. Thus, we conclude that 5-InsP7regulates p53 expression, whereas 1,5-InsP8regulates ATP levels. These findings attribute hitherto unsuspected functionality for 1,5-InsP8to bioenergetic homeostasis.