Metastasis of human gastric adenocarcinoma partly depends on phosphoinositide-specific phospholipase γ1 expression

Abstract

Abstract:It is known that phosphoinositide-specific phospholipases g1 (PLCg1) can trigger several signalling pathways to regulate cell proliferation, differentiation, and metastasis. However, whether this kinase is highly expressive and active in human gastric adenocarcinomas, and whether it can play an important role in the development of the cancer, have not yet been investigated. The aim of the study was to investigate the expression of PLCg1 in human gastric adenocarcinoma, while the question of whether PLCg1 can be activated through protein kinase B (Akt) signalling pathways to regulate cell migration was further explored using human gastric adenocarcinoma BGC-823 cell line. The expression of PLCg1 in human adenocarcinoma was detected using immunohistochemical staining. The BGC-823 cells were cultured and treated with inhibitors or transfected with plasmid construction. The cell migration of BGC-823 cells was measured with wound healing assay, cell migration assay, and the ruffling assay. The expression levels of PLCg1 and its related signal molecules in BGC-823 cells were assessed using Western blot analysis or gelatine zymography assay. PLCg1 was highly expressed in human gastric adenocarcinomas, especially in the region with lymph node metastasis. It was shown that migration of BGC-823 cells in vitro depends on PLCg1 activation. This activation is mediated through Akt, an upstream of PLCg1 that triggers the PLCg1/extracellular signal-regulated kinase (ERK)/matrix metalloproteinase (MMP) pathway in BGC-823 cells. PLCg1 activities play an important role in the metastasis of gastric adenocarcinoma, and may serve as a potential therapeutic target in this type of cancer.

Phosphoinositide specific phospholipase Cγ1 inhibition-driven autophagy caused cell death in human lung adenocarcinoma A549 cells in vivo and in vitro

“…After cells were tranduced with the different shPLCγ1 vectors by a lentiviral transfection strategy, stable cell lines were obtained under the pressure of puromycin (2 μg/mL, BioVision, Inc., Milpitas, CA, USA). In addition, cells were transiently transfected for 24 hours with pRK5-HA-PLCγ1 (pRK5-PLCγ1) vector expressing PLCγ1, p3xFLAG-CMV10-AMPKα1 (AMPKα1) vector expressing AMPKα1, p3xFLAG-CMV10-AMPKα2 (AMPKα2) vector expressing AMPKα2, using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA), following the manufacturer's procedure and previous studies [9,41].…”

“…Cells were seeded in 96-well plates and each well contained 100 μL of complete growth medium and cultured for the indicated time. The number of viable cells were measured by 3-(4, 5-Dimethylthiazol-2-y)-2,5-diphenyl-tetrazolium bromide (MTT) assay as described as previous studies [9,42].…”

“…According to the previous studies [9,43], cells in serum-free RPMI-1640 were placed into the top chambers of Transwell inserts set with 8 μm pore filters and RPMI-1640 complete medium was added to the bottom chamber. After 24 hours, cells on top of the membrane were removed with a cotton swab, and then the membrane of Transwell inserts was fixed with 4% paraformaldehyde.…”

“…Phosphoinositide specific phospholipase Cγ1 (PLCγ1) plays an important role in regulating cell proliferation and migration of tumor [8][9][10]. Very recent study reports that PLCG1 could act as an oncogene in hepatocellular carcinoma carcinogenesis and may serve as a valuable prognostic marker and potential therapeutic target for hepatocellular carcinoma [11].…”

DAG/PKCδ and IP3/Ca2+/CaMK IIβ Operate in Parallel to Each Other in PLCγ1-Driven Cell Proliferation and Migration of Human Gastric Adenocarcinoma Cells, through Akt/mTOR/S6 Pathway

“…PLCγ mediates high levels of glucose and insulin-induced cell proliferation and migration in MDA-MB-468 breast cancer and SW480 colon cancer cells in vitro [ 8 ]. Our previous study also showed the higher expression of PLCγ1 in human gastric adenocarcinoma tissue and that the metastasis of human gastric adenocarcinoma cells partly depends on PLCγ1 expression [ 9 ]. Moreover, it has been shown that the depletion of PLCγ expression or inhibition of its activity not only significantly increases cisplatin-induced apoptosis but also suppresses the invasive ability of RhoGDI2-overexpressing SNU-484 gastric cancer cells [ 10 ].…”