Gene Expression Profiling of Lung Squamous Cancer Stratified by NSD3 Expression

NSD3 is located on a region of chromosome 8 (8p11-12) that is frequently amplified in several forms of cancer. It has been suggested that NSD3 (WHSC1L1) may be a driver of tumorigenesis in this context (Mahmood et al. 2013). However, it is still unclear how amplification of NSD3 contributes to the disease and whether or not it Read More …

NSD3’s Role in Promoting Epithelial to Mesenchymal Transition is Isoform Dependent

To understand what proteins do, scientists often test the consequence their absence has on a cell. A common method uses a cells regulatory machinery to disrupt gene expression with complementary RNA molecules, known as RNA interference. Here I used small interfering RNAs (siRNAs) to deplete NSD3 and test the effect this has on epithelial cell identity in Read More …

Following Up on NSD3’s Involvement in Epithelial to Mesenchymal Reprogramming

In my last blog post, I observed morphological changes that were indicative of an epithelial to mesenchymal transition (EMT) when NSD3short was overexpressed from a stably integrated transgene in the cancer cell line H1299. Importantly, NSD3 has been identified as amplified in a number of cancer types, including lung (Figure 1) (cBioPortal: Cerami et al., Cancer Discov. 2012 Read More …

Over-expression of the NSD3-Short Isoform in H1299 Lung Cancer Cells Alters Cellular Morphology

Recently, a study of cancer-focused protein-protein interactions identified a direct interaction between NSD3 and the oncoprotein Myc in the non-small cell lung cancer cell line H1299 (Li, Z et al. 2017).  Importantly, the study found that increased NSD3 expression levels can activate Myc in H1299 cells as measured by an E-box luciferase reporter. However, the functional significance of Read More …

Western blots & Lentiviruses: Building Tools to Study NSD3

The early stages of a project often involve spending considerable time building tools to effectively tackle the problem. In cell biology, this often includes testing antibodies and expression constructs in your cell lines of interest. Putting in the work to properly validate these reagents early on in a study is critical. Poorly validated antibodies are, at least Read More …

Cloning an NSD3-Short-3xFLAG Construct into a Lentiviral Expression System

Antibodies are fundamental tools in the exploration of cellular biology. Unfortunately, there is a lack of high-quality NSD3 antibodies available. To overcome this, I have generated a bicistronic NSD3-Short-3xFLAG – IRES – PuroR lentiviral expression plasmid for the purpose of creating cell lines that stably express 3xFLAG-tagged NSD3-short. This will allow me to use a Read More …

Project Overview: Understanding Enhancer Biology in Cancer

Every cell in your body contains the exact same set of genetic instructions encoded in its genome. However, for complex multicellular organisms such as ourselves, we depend on many specialized cell types that exhibit strikingly different features. This is largely accomplished by precisely controlling how and when genetic information is read out in each cell, Read More …