Expression of α-fetoprotein in gastric cancer AGS cells contributes to invasion and metastasis by influencing anoikis sensitivity

Abstract

α-fetoprotein (AFP) is a valuable tumor marker for many types of cancers, including primary gastric cancer (GC). However, the effects of AFP expression on the metastasis and anoikis sensitivity of GC remain unclear. The present study aimed to explore the role and possible mechanism of AFP in the invasion and metastasis of GC AGS cells, particularly in the anoikis sensitivity of AGS cells. In the present study, the expression of AFP in cultured AGS cells was assayed firstly by RT-PCR, western blotting and sequencing. Then, a specific AFP siRNA was applied to interfere with AFP expression and poly(2-hydroxyethyl methacrylate) (poly-HEMA) was used to block cell anchorage. The invasion and metastatic ability, and anoikis sensitivity detections were conducted based on Transwell chamber assay, anoikis assay kit and western blotting. Our results confirmed the expression of AFP in AGS cells. Then, we found that interference of AFP with siRNA attenuated the invasion and metastasis of AGS cells and induced a significant upregulation of E-cadherin and downregulation of N-cadherin expression (P<0.05). Cell apoptosis and anoikis were induced when cell anchorage was blocked by poly-HEMA treatment, which was exacerbated significantly when cells were exposed to AFP siRNA. Moreover, interference of AFP when cell anchorage was blocked enhanced the expression of the pro-apoptotic proteins Bax, caspase-3 and -9, and decreased the expression of the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2). In conclusion, the present study demonstrated that interference of AFP reduced AGS cell invasion and metastasis by enhancing anoikis sensitivity. The present study provides new insight for the treatment of GC and suggests AFP as a potential therapeutic target by regulating anoikis sensitivity.

ER stress dependent microparticles derived from smooth muscle cells promote endothelial dysfunction during thoracic aortic aneurysm and dissection

“…EC anoikis was detected using a CytoSelect™ 96-Well Anoikis Assay kit (Cell Biolab) according to the manufacturer’s protocol [15–17]. The kit allows the quantitation and monitoring of anoikis in cells using a precoated hydrogel plate.…”

Inhibition of apoptosis by miR‑122‑5p in α‑fetoprotein‑producing gastric cancer

“…In the present study, we confirmed that the expression level of miR-122-5p was significantly increased in the AFPGC cell line than in common gastric cancer (GC) cell lines, and also inhibition of miR-122-5p significantly reduced AFP levels in the AFPGC cell line. Conversely, several studies have clearly demonstrated that the AFP molecule itself directly contributes to the promotion of tumor progression (13)(14)(15). With regard to these findings, we hypothesized that miR-122-5p may directly regulate tumor progression in AFPGC.…”

“…Current functional analyses by the gene transfection approach clearly demonstrated that inhibition of miR-122-5p significantly reduced proliferation in AFPGC cells through induction of apoptosis. Several previous studies have revealed that resistance to apoptosis could play a crucial role in the aggressive characteristics of AFPGC (14,15). Apoptotic resistance may be a major cause of the low chemosensitivity in AFPGC.…”

HCRP-1 regulates EGFR–AKT–BIM-mediated anoikis resistance and serves as a prognostic marker in human colon cancer

“…The results showed that p-AKT, p-FoxO3a and BIM expression, and anoikis resistance were reversed by these pretreatments in HCT116 and SW620 cells. As anoikis is important for disease and tumor metastasis 31 , we also carried out animal experiments. The results showed that reduced HCRP-1 expression promoted tumor metastasis and activated the EGFR and AKT signaling pathway, which were consistent with the results in vitro.…”