Effect of combined use of Smo inhibitors and ptch1 inhibitors in hedgehog pathway on in vitro cell proliferation and invasion

Cancer is a disease with the highest mortality and incidence worldwide. Although the mechanisms of cancer are being clarified day by day, there are still many unknowns about the disease. Clarifying the signaling pathways will increase the knowledge about cancer and may guide the diagnosis and treatment of cancer. The hedgehog signaling pathway is involved in cell proliferation, cell cycle, tissue regeneration, wound healing, and homeostasis during human embryogenesis. Abnormal Sonic Hedgehog (Shh) activation in adulthood has been shown to play an important role in neoplastic growth, development of cancer stem cells, and epithelial-mesenchymal transition processes. In the absence of hedgehog signaling, Patched 1 (Ptch1), a member of the 12-transmembrane receptor family, acts catalytically to inhibit the activity of the 7-transmembrane protein Smoothened (Smo) to prevent its localization to the cell membrane. Hedgehog signaling pathway activation causes changes in cilia in the cell membrane. Ptch abrogates the inhibition on Smo and Smo is sequentially phosphorylated. After phosphorylations, the activated Gli1 transcription factor is translocated to the cell nucleus and the target genes that will respond to the hedgehog signal are transcribed. This expression of Gli1 is directly and indirectly related to cell proliferation. Inhibition of the hedgehog signaling pathway with specific inhibitors and antagonists is considered an innovative way of cancer therapy. In this project, the presence of hedgehog signaling pathway was demonstrated in commercial glioblastoma cell line U87-MG, and the effect of combined use of Ptch1 antagonist Robotnikinin and Smo inhibitor Vismodegib on the hedgehog signaling pathway was investigated. In order to observe this effect, cell proliferation, migration and invasion analyzes were performed. In addition, in silico comparisons of the antagonist and inhibitor used were made and it was concluded that robotnikin could be a drug-like molecule. The results obtained from in vitro studies show that effective results can be obtained with combined treatment. It has been observed that the combined use of agents in combined treatments is more effective than their use alone. This is the first study in the literature to apply combined treatments of Smo and Ptch1, two proteins of the hedgehog signaling pathway, with vismodegib and robotnikinin.