Effect of nicastrin on hepatocellular carcinoma proliferation and apoptosis through PI3K/AKT signalling pathway modulation

Abstract

Background: Increasing evidence has proven that the γ-secretase complex plays significant roles in the carcinogenesis of malignancies. However, the independent effect of nicastrin (NCSTN), the largest constituent of the γ-secretase complex, on the progression of hepatocellular carcinoma (HCC) remains to be discovered. Methods:In our study, we used open online databases, including the Oncomine database, GEPIA and KMPlotter, to analyse the expression of 4 genes and their correlation with prognosis in HCC. NCSTN expression in 60 HCC patients from our centre was determined by immunohistochemical staining and qRT-PCR. The clinical and prognostic significance of NCSTN expression were analysed statistically. Stable Sk-hep1 cell lines with NCSTN overexpression were established using lentivirus-based vectors, and RNAi technology was used to transiently downregulate NCSTN expression in HepG2 cell lines. Cell growth and apoptosis were assessed by using EdU, clone formation, flow cytometry and Western blotting assays.Results: Bioinformatics analysis showed that NCSTN mRNA expression was generally higher in HCC tissues than in normal tissues according to a meta-analysis of 9 HCC datasets, excluding PS-1, PEN-2 and APH-1. Moreover, NCSTN was associated with a poor prognosis in HCC patients from The Cancer Genome Atlas (TCGA). Although the relationship between NCSTN levels and the clinicopathological features of HCC patients was not significant, a survival analysis of HCC patients from TCGA indicated that overall and disease-free survival were significantly associated with NCSTN expression. NCSTN expression in HCC cell lines regulated cell growth and apoptosis in vitro. NCSTN downregulation in HepG2 cells inhibited tumour growth ability in vivo. In addition, NCSTN downregulation in HepG2 cell lines decreased p-PI3K and p-Akt expression, and IGF1, a PI3K/Akt activator, neutralized the effects on PI3K and Akt phosphorylation. Moreover, NCSTN overexpression in Sk-hep1 cells activated the PI3K/Akt signalling pathway, and MK-2206, a PI3K/Akt inhibitor, reversed this activation according to Western blotting analysis. Conclusions:We suggest that NCSTN serves as an oncogene in HCC by promoting growth and inhibiting apoptosis via the PI3K/Akt pathway, providing a potential novel therapeutic target for HCC treatment.

Proteomics-based investigation of cerebrovascular molecular mechanisms in cerebral amyloid angiopathy by the FFPE-LMD-PCT-SWATH method

“…For AKT signalling (Fig. 5 ), we found the decreased expressions of NCSTN [ 43 ], TPD52L2 [ 44 ], PSMC2 [ 45 ], FKBP4 [ 46 ] and ARHGAP1 [ 47 ] which activate AKT signalling (Fig. 4 ).…”

Effect of astragaloside on diaphragm cell apoptosis in chronic obstructive pulmonary disease

“…AKT is a clinically common serine/threonine‐specific protein kinase, and its phosphorylated form is p‐AKT, which shows biological activity. With homology to protein kinases A and C, AKT is also known as protein kinase B (PKB) (Ma et al., 2020; Wang et al., 2020). p‐Akt is a cell cycle regulator, proliferation promoter, and apoptosis inhibitor by acting on its downstream molecules.…”

LY‑294002 enhances the chemosensitivity of liver cancer to oxaliplatin by blocking the PI3K/AKT/HIF‑1α pathway

“…MK-2206 inhibits the autophosphorylation of AKT at threonine 308 and serine 473 (21). Preclinical studies have shown that MK-2206 has an inhibitory effect on human cancer cell lines, such as breast, lung, colon and liver cancer (22)(23)(24)(25). PI3K/AKT/HIF-1α reportedly induces chemoresistance in liver cancer cells (26,27), and LY-294002 can enhance the chemosensitivity of liver cancer (28).…”