Ancer organoids expressing high levels of GLI2 had been found to be very resistant to chemotherapeutic drugs epirubicin, oxaliplatin, and 5fluorouracil in comparison to these without the need of GLI2 expression in both in vitro and in vivo, although therapy with GANT61 resensitized the organoids to chemotherapy [103]. Additional study by the same group revealed that the PI3K/AKT/mTOR pathway activated in patientderived organoids noncanonically upregulated GLI1 and GLI2 to induce PDL1 expression, which may very well be properly suppressed with rapamycin [104]. Taken with each other, GLI1/2 mediates mTORinduced PDL1 expression to promote immune evasion of cancer cells, also as promotes chemoresistance. Kasiri et al. reported a trend exactly where higher GLI1 transcript expression in NSCLC individuals was related with worse OS [105]. In addition, the loss of GLI1 by shRNAmediated knockdown significantly suppressed cell proliferation of subcutaneous SCC xenograft tumors in vivo. In vitro study demonstrated that inhibition of PI3K/mTOR signaling effectively diminished GLI1 expression and inhibited clonogenicity and proliferation of lung squamous cell carcinoma cell lines. Likewise, regulation of GLI1 was also independent of canonical Hh signaling, as neither SMO inhibition by Triclabendazole sulfoxide medchemexpress GDC0449 nor induction by SAG had a substantial effect on both GLI1 transcript and protein levels; SMO inhibition also did not have an effect on colony formation and cell proliferation. The concurrent inhibition of GLI and PI3K/AKT/mTOR signaling demonstrated a synergistic effect in inhibiting in vivo cancer cell growth, evident by decreased tumor burden of xenografts compared to treatment with any of the agents alone [105]. GLI1 expression has also been reported to become regulated by PI3K/AKT/mTOR signaling in several other cancers to market tumorigenesis, which includes esophageal adenocarcinoma [106], melanoma [113], osteosarcoma [107], pancreatic cancer, ovarian cancer [108], and renal cancer [109]. S6K1/2, members from the ribosomal S6 kinase family, are downstream targets of PI3K/AKT/mTOR and are involved in protein synthesis and cell proliferation. Notably, their activation has been linked to elevated GLI1 expression and activity in numerous cancers. Tumor necrosis factoralpha (TNF) induced SK61 phosphorylation, which was associated with enhanced GLI1 expression and GLI1 target genes, such as cell cycle regulators CCND1 and nMyc, in prostate cancer PC3 cells. Constant with the upregulation of these genes, GLI1 depletion by either GANT61 or siRNAmediated knockdown correctly suppressed PC3 cell viability, liquid colony formation, and cell proliferation. Conversely,Biomedicines 2021, 9,26 ofgenetic and pharmacological inhibition of PI3K/mTOR inhibited TNFinduced SK61 phosphorylation and consequently GLI1 expression, which led to decreased PC3 cell viability [110]. Interestingly, a study by Wang et al. reported that in esophageal adenocarcinoma cell lines, TNF stimulation and ectopic SK61 expression regulate GLI1 activity by phosphorylation of its Ser84 residue, thereby dissociating GLI1 from SUFU and enabling GLI1 translocation in to the nucleus [111]. Conversely, inhibition of SK61 activation by PI3K/mTOR inhibitor rapamycin and RAD001 enhanced HH inhibitor GDC0449 cytotoxic impact in each in vitro and in vivo models. Furthermore, GLI1 was expected for TNF3/mTOR/S6K1mediated cell proliferation, as GLI1 knockdown abrogated TNFand S6K1induced cell viability, proliferation, and invasion. Of note, SMO inhibition with cyclopa.