The Natural Alkaloid Piperlongumine Inhibits Metastatic Activity and Epithelial-to-Mesenchymal Transition of Triple-Negative Mammary Carcinoma Cells.

In this study, we determined the effect of low dose piperlongumine on the motility/invasive capacity and epithelial-to-mesenchymal transition (EMT) of MDA-MB-231 triple-negative breast cancer (TNBC) cells and the metastasis of 4T1 mouse mammary carcinoma cells. MTT assays measured the effect of piperlongumine on TNBC cell growth. Motility/invasiveness were determined by gap closure/transwell assays. Western blotting assessed ZEB1, Slug, and matrix metalloproteinase (MMP) 9 expression. Interleukin (IL) 6 was detected by ELISA. MMP2, E-cadherin, and miR-200c expression was determined by real-time quantitative polymerase chain reaction. Reactive oxygen species (ROS) were measured by flow cytometry. The orthotopic 4T1 mouse model of breast cancer was used to examine metastasis. Piperlongumine-treated MDA-MB-231 cells showed reduced motility/invasiveness, decreased MMP2 and MMP9 expression, increased miR-200c expression, reduced IL-6 synthesis, decreased expression of ZEB1 and Slug, increased E-cadherin expression, and epithelial-like morphology. Piperlongumine also inhibited transforming growth factor ß-induced ZEB1 and Slug expression. ROS accumulated in piperlongumine-treated cells, while changes in metastasis-associated gene expression were ablated by exogenous glutathione. Metastasis of 4T1 cells to the lungs of BALB/c mice was dramatically reduced in piperlongumine-treated animals. These findings reveal a previously unknown capacity of low dose piperlongumine to interfere with TNBC metastasis via an oxidative stress-dependent mechanism.

An Automated Text Messaging System (Tonsil-Text-To-Me) to Improve Tonsillectomy Perioperative Experience: Exploratory Qualitative Usability and Feasibility Study.

BACKGROUND: Inexperience and forgetting perioperative care instruction are significant drivers of parental stress during pediatric tonsillectomy care. With the widespread use of mobile technology, parents now desire a system that provides them with information that is timely, accessible, and comprehensive. Tonsil-Text-To-Me (TTTM) is a text messaging system that sends out automated and timed texts to parents of children who are undergoing tonsillectomy. OBJECTIVE: The objective of this study was to pilot-test TTTM to assess for feasibility and usability and collect suggestions for system improvements desired by parents from a pediatric otolaryngology text message service. METHODS: Parents of pediatric patients who were being scheduled for tonsillectomy with or without adenoidectomy were prospectively enrolled. An exploratory qualitative study using a semistructured interview guide was performed after parents received the automated texts 2 weeks before and 1 week after their child's surgery. RESULTS: A total of 7 parents were interviewed (data saturation was reached). Participants were all of maternal relation to the patient. Overall, all parents felt that the TTTM service was an improvement to the current standard model of information delivery. Parents also reported that the text messages reduced their anxiety and improved their performance when caring for their children during the perioperative period. No parents expressed privacy concerns about receiving texts and regarding the information included in the messages. Service suggestions showed that parents were eager for more information and had a high threshold for message reception regarding their child's surgical care. CONCLUSIONS: All parents expressed enthusiasm for a text message service during their child's tonsillectomy perioperative period. The care instructions and reminders provided to parents via automated and timed text messages may be a strategy to improve information delivery in a simple and accessible format that could empower families in their own health care.

Characterization of cancer stem cell drug resistance in the human colorectal cancer cell lines HCT116 and SW480.

BACKGROUND: Cancer stem cells (CSCs) share a number of properties with somatic stem cells including heightened protective mechanisms and the ability to self-renew. CSCs are a critical subpopulation of cancer cells implicated in tumor formation, metastases and recurrence. METHODS: We used serial colonosphere culture to enrich for CSCs from two human CRC cell lines. The expression of proposed colorectal CSC markers and multi-drug resistance genes were assessed via flow cytometry and RT-qPCR. Drug resistance gene expression and self-renewal ability were also determined following treatment with the chemotherapeutic 5-fluorouracil. RESULTS: Colonosphere culture successfully enriched for a subpopulation of cells with CSC-related gene expression and heightened self-renewal ability, particularly in the SW480 cell line. Chemotherapy treatment significantly reduced sphere formation however a small fraction of cells survived treatment and retained their self-renewal ability. CONCLUSIONS: Our findings support the use of the sphere formation assay to study CSCs. The ability of cells to self-renew following chemotherapy treatment highlights the importance of targeting both the bulk of tumor cells and the CSC population to prevent recurrence in colorectal cancer.

The effects of folic acid on global DNA methylation and colonosphere formation in colon cancer cell lines.

Folate and its synthetic form, folic acid (FA), are essential vitamins for the regeneration of S-adenosyl methionine molecules, thereby maintaining adequate cellular methylation. The deregulation of DNA methylation is a contributing factor to carcinogenesis, as alterations in genetic methylation may contribute to stem cell reprogramming and dedifferentiation processes that lead to a cancer stem cell (CSC) phenotype. Here, we investigate the potential effects of FA exposure on DNA methylation and colonosphere formation in cultured human colorectal cancer (CRC) cell lines. We show for the first time that HCT116, LS174T, and SW480 cells grown without adequate FA demonstrate significantly impaired colonosphere forming ability with limited changes in CD133, CD166, and EpCAM surface expression. These differences were accompanied by concomitant changes to DNA methyltransferase (DNMT) enzyme expression and DNA methylation levels, which varied depending on cell line. Taken together, these results demonstrate an interaction between FA metabolism and CSC phenotype in vitro and help elucidate a connection between supplemental FA intake and CRC development.