DKK3 knockdown confers negative effects on the malignant potency of head and neck squamous cell carcinoma cells via the PI3K/Akt and MAPK signaling pathways

Abstract

Dickkopf-related protein 3 (DKK3), which is a member of the Dickkopf WNT signaling pathway inhibitor family, is considered to be a tumor suppressor, due to its reduced expression in cancer cells and its ability to induce apoptosis when overexpressed by adenovirus. However, our previous study demonstrated alternative functions for DKK3 in head and neck squamous cell carcinoma (HNSCC). Our study reported that DKK3 expression was predominantly upregulated in HNSCC cell lines and tissue samples, and its expression was significantly correlated with poor prognosis. Furthermore, DKK3 overexpression in HNSCC cells significantly increased cancer cell proliferation, migration, invasion and in vivo tumor growth. These data have led to the hypothesis that DKK3 may exert oncogenic functions and may increase the malignant properties of HNSCC. The present study established a stable DKK3 knockdown cell line (HSC-3 shDKK3) using lentivirus-mediated short hairpin RNA, and assessed its effects on cancer cell behavior using MTT, migration and invasion assays. In addition, its effects on in vivo tumor growth were assessed using a xenograft model. Furthermore, the molecular mechanisms underlying the effects of DKK3 knockdown were investigated by microarray analysis, pathway analysis and western blotting. Compared with control cells, HSC-3 shDKK3 cells exhibited significantly reduced proliferation, migration and invasion, and formed significantly smaller tumor masses when subcutaneously transplanted into nude mice. In addition, in HSC-3 shDKK3 cells, the expression levels of phosphorylated (p)-protein kinase B (Akt) (Ser473), p-phosphoinositide 3-kinase (PI3K) p85 (Tyr467), p-PI3K p55 (Try199), p-3-phosphoinositide-dependent protein kinase-1 (PDK1) (Ser241) and total p38 mitogen-activated protein kinase (MAPK) were reduced. Furthermore, phosphorylation of mechanistic target of rapamycin (mTOR) (Ser2448) was slightly decreased in HSC-3 shDKK3 cells, which may be due to the increased expression of DEP domain-containing mTOR-interacting protein. Conversely, DKK3 overexpression in HSC-3 shDKK3 cells rescued cellular proliferation, migration and invasion. With regards to expression levels, p-PI3K and p-PDK1 expression was not altered, whereas mTOR and p-p38 MAPK expression was elevated. These data supported the hypothesis and indicated that DKK3 may contribute to the malignant phenotype of HNSCC cells via the PI3K/Akt/mTOR and MAPK signaling pathways.

Anethole induces anti-oral cancer activity by triggering apoptosis, autophagy and oxidative stress and by modulation of multiple signaling pathways

“…Inhibition of these pathways can be an excellent target for cancer therapy. In addition, MAPK and Wnt pathways activated are a marker of poor prognosis in human oral cancer, and persistent MAPK activation in it is associated with enhanced cellular and tumor growth 31 , 32 . The high cytotoxic effect of anethole in oral cancer cells at low concentration but in normal gingival cells, it has a marginal effect at low concentration, but this molecule appears to have some toxicity when used at high concentration.…”

Targeting AKT/mTOR in Oral Cancer: Mechanisms and Advances in Clinical Trials

“…Several studies also demonstrated that proteins such as the receptor for activated C kinase 1 (RACK1), dickkopf-related protein 3 (DKK3) and T-cadherin are crucial for the proliferation and survival of OSCC. These proteins regulate the proliferation and survival of OSCC via the activation of the AKT/mTOR pathway [71,110,111]. Besides, keratin 17 also displayed high expression in OC and this protein promoted tumorigenesis via the stimulation of AKT/mTOR signaling and glucose uptake [67].…”

Hinokiflavone Inhibits Growth of Esophageal Squamous Cancer By Inducing Apoptosis via Regulation of the PI3K/AKT/mTOR Signaling Pathway

“…This suggested that HF can regulate post-translational modification of PI3K by physically interacting with the protein, as shown by the results of the GO enrichment, KEGG pathway, and molecular docking analyses. It is plausible that HF may directly attenuate the phosphorylation of PI3K at tyrosine 199, as reported in another study (40). The PI3K/ Akt/mTOR pathway is a crucial signaling cascade in living organisms that has to be activated in a variety of cancers and to regulate cell proliferation, invasion, and migration (20,41).…”