TTC36 inactivation induce malignant properties via Wnt-β-catenin pathway in gastric carcinoma

Abstract

Objective: Tetratricopeptide repeat (TRP)-mediated cofactor proteins are involved in a wide range of cancers. TTC36 is little studied member of TRP subfamily. This study aimed to investigate the role of TTC36 in human gastric carcinoma (GC) and explore the potential underlying mechanisms. Methods: The analysis of TTC36 differential expression in GC was conducted using data from TCGA and a human tissue microarray. And effects of TTC36 expression on the prognosis of patients with gastric carcinoma were analyzed using the data from the GEO database. Lentivirus was transfected into the cell lines of AGS and BGC823 to construct overexpression and knocked down TTC36 cell model respectively. The effect of TTC36 expression on the growth, apoptosis and cell cycle of cells was explored in vitro. Downstream molecules were detected by western blotting. GSEA predicted signal pathway and related proteins were then detected. Results: TTC36 expression in human GC tissues was found significantly lower than that in adjacent normal tissues and closely related to clinical prognosis. The overexpression of TTC36 notably inhibited tumor progression, cell cycle G1/S transfer and increased apoptosis in AGS cells. Conversely, the opposite effects were observed when TTC36 was suppressed in BGC823 cells. The expression of cleaved caspase3, Survivin, cyclin D1 and c-Myc were consistent with the phenotype in TTC36 operated GC cell lines. Intriguingly, GSEA analysis predicted Wnt-β-catenin pathway involved in TTC36 induced effects in GC cells, expression of β-catenin and downstream molecules such as TCF4, c-jun and pAKT were found negative related to TTC36 expression in GC cells. Notably, treatment with the Wnt/β-catenin inhibitor XAV939 dramatically attenuated the effects of TTC36 in GC cells. Conclusion: These results signify a critical role for TTC36 as a tumor suppressor in gastric carcinoma via regulating Wnt-β-catenin pathway.

LATS1 exerts tumor suppressor functions via targeting Gli1 in colorectal cancer

“…IHC staining of paraffin-embedded human tissues was carried out as reported previously 25 , 26 . In brief, Samples were dewaxed, rehydrated and endogenous peroxidase activity was blocked with 3% hydrogen dioxide.…”

“…The Lentivirus vectors carrying LATS1 overexpression or short hairpin RNA (shRNA) targeting LATS1 and their control vectors were invented by Genechem Company (Shanghai, China). We performed the transfection following the company's instructions, which were documented in our earlier study 25 , 26 . Lipofectamine RNAiMax (Invitrogen) was used to transfect small interfering RNA (siRNA) at 20 nM of final concentration, as directed by the manufacturer.…”

“…The bands were quantified using the Imaging J analysis software. The detailed protocol was described previously 25 , 26 . Following antibodies used in this study: anti-LATS1 (dilution 1:1000; #17049-1-AP, Proteintech), anti-Gli1 (dilution 1:1000; #ab217326, Abcam), anti-YAP1 (dilution 1:1000; #14074, Cell Signaling Technology (CST)), anti-p-mTOR (dilution 1:1000; #9205, CST), anti-mTOR (dilution 1:1000; #2983, CST), anti-GAPDH (dilution 1:5000; #AG019, Beyotime).…”

“…Smoothened (SMO), known as the canonical Hedgehog pathway, can activate Gli1, a major transcription factor in the Hedgehog signaling pathway 24 . It can also be activated by protein kinase B (AKT), extracellular signal-regulated kinase (ERK), and mammalian target of rapamycin (mTOR) which is known as the non-canonical Hedgehog pathway 24 , 25 .…”

Mecanismos epigenéticos em hepatoblastomas: análise do transcriptoma e sua regulação por metilação de DNA

“…TTC36 downregulation evidenced in HBL was associated with alterations in the metabolism of amine and small molecules, development process and cell differentiation. This gene is associated with tumoral suppressor activities, and its inactivation in gastric cancer might promote cell proliferation, at least in part, through activating the Wnt/β-catenin signaling pathway 193 . Glucokinase (GCK) contributes to the reprogramming of energy metabolism in cancer cells 194 .…”