Supplemental Table 1 and 2 and Supplemental Figure 1-5. from Activin Upregulation by NF-κB Is Required to Maintain Mesenchymal Features of Cancer Stem–like Cells in Non–Small Cell Lung Cancer

Wamsley, J. Jacob; Kumar, Manish; Allison, David F.; Clift, Sheena H.; Holzknecht, Caitlyn M.; Szymura, Szymon J.; Hoang, Stephen A.; Xu, Xiaojiang; Moskaluk, Christopher A.; Jones, David R.; Bekiranov, Stefan; Mayo, Marty W.

doi:10.1158/0008-5472.22401968.v1

Supplemental Table 1 and 2 and Supplemental Figure 1-5. from Activin Upregulation by NF-κB Is Required to Maintain Mesenchymal Features of Cancer Stem–like Cells in Non–Small Cell Lung Cancer

journal contribution

posted on 2023-03-30, 22:30 authored by J. Jacob Wamsley, Manish Kumar, David F. Allison, Sheena H. Clift, Caitlyn M. Holzknecht, Szymon J. Szymura, Stephen A. Hoang, Xiaojiang Xu, Christopher A. Moskaluk, David R. Jones, Stefan Bekiranov, Marty W. Mayo

Supplemental Table 1 and 2 and Supplemental Figure 1-5. Supplemental Table 1: Oligonucleotide sequences of shRNA molecules targeting INHBA/Activin and the targeted sequences. Supplemental Table 2: Oligonucleotide sequences used for amplification in QRTPCR. Supplemental Figure 1: Microarray analysis performed on mesenchymal A549 cells show massive and reproducible epigenetic changes in gene expression without upregulating TGFβ or NODAL. Supplemental Figure 2: Mesechymal NSCLC cells are capable of sustaining INHBA upregulation over a four-day time course. Supplemental Figure 3: The ability of condition media to stimulate SNAI1 transcription is blocked by the ALK4/ALK5/ALK7 inhibitor. Supplemental Figure 4: Infection of A549 cells with shRNA targeting INHBA. Supplemental Figure 5: Representative images of transwell assay inserts.

History

ARTICLE ABSTRACT

Soluble growth factors and cytokines within the tumor microenvironment aid in the induction of the epithelial-to-mesenchymal transition (EMT). Although EMT promotes the development of cancer-initiating cells (CIC), cellular mechanisms by which cancer cells maintain mesenchymal phenotypes remain poorly understood. Work presented here indicates that induction of EMT stimulates non–small cell lung cancer (NSCLC) to secrete soluble factors that function in an autocrine fashion. Using gene expression profiling of all annotated and predicted secreted gene products, we find that NF-κB activity is required to upregulate INHBA/Activin, a morphogen in the TGFβ superfamily. INHBA is capable of inducing and maintaining mesenchymal phenotypes, including the expression of EMT master-switch regulators and self-renewal factors that sustain CIC phenotypes and promote lung metastasis. Our work demonstrates that INHBA mRNA and protein expression are commonly elevated in primary human NSCLC and provide evidence that INHBA is a critical autocrine factor that maintains mesenchymal properties of CICs to promote metastasis in NSCLC. Cancer Res; 75(2); 426–35. ©2014 AACR.