siRNA-mediated silencing of bFGF gene inhibits the proliferation, migration, and invasion of human pituitary adenoma cells

Abstract

Human pituitary adenoma is one of the most common intracranial tumors with an incidence as high as 16.7%. Recent evidence has hinted a relationship between growth factors of pituitary or hypothalamic origin and proliferation of human pituitary adenoma cells. This study explores the effects of small interfering RNA-mediated silencing of basic fibroblast growth factor gene on the proliferation, migration, and invasion of human pituitary adenoma cells. Human pituitary adenoma tissues were collected to obtain human pituitary adenoma cells. The basic fibroblast growth factor silencing interference plasmid was constructed, and the human pituitary adenoma cells were transfected and assigned as basic fibroblast growth factor-small interfering RNA, negative control-small interfering RNA, and blank groups. The quantitative real-time polymerase chain reaction and Western blotting were carried out to detect the expression of basic fibroblast growth factor, pituitary tumor transforming gene, vascular endothelial growth factor, cluster of differentiation 147, and matrix metalloproteinase 9. Cell Counting Kit-8 assay was conducted to observe cell proliferation at 0, 24, 48, and 72 h. Flow cytometry was used to determine cell cycle. Transwell and scratch test were applied to detect the invasion and migration of pituitary adenoma cells. Protein kinase C activity was detected. In comparison with the blank group, the basic fibroblast growth factor-small interfering RNA group showed reduced messenger RNA and protein expression of basic fibroblast growth factor, reduced cell viability at 24, 48, and 72 h, increased cells in G0/G1 stage, declined cells in S and G2/M stages, decreased number of cell migration, shortened migrating distance, reduced protein kinase C activity, and decreased expression of pituitary tumor transforming gene, vascular endothelial growth factor, cluster of differentiation 147, and matrix metalloproteinase 9. However, the negative control-small interfering RNA group had no evident differences in basic fibroblast growth factor expression, cell viability, cell cycle, number of cell migration, migrating distance, protein kinase C activity, and expression of pituitary tumor transforming gene, vascular endothelial growth factor, cluster of differentiation 147, and matrix metalloproteinase 9 compared with the blank group. The study provides evidence that small interfering RNA-mediated silencing of basic fibroblast growth factor gene inhibits the proliferation, migration, and invasion of human pituitary adenoma cells.

Endocan, VEGF, FGF2 and PDGF Expression in Pituitary Tumors. A Precise Angiogenic Marker Relationship in the Context of Tumor Invasiveness

“…Interestingly, Almong et al described endocan was markedly expressed in concordance with the switch between dormant to fast growing phenotype in experimental angiogenic tumors (15). Moreover, the enhanced FGF2 expression we found in ESM1 positive tumors could be supporting the fast-growing state in these tumors as FGF2 has been linked to invasion in cancer and in pituitary adenomas as well (35,43).…”

“…Moreover, the knockdown of Esm1 in HUVEC endothelial cells inhibited tube formation and increased sensitivity to the anti-angiogenic Avastin treatment in vitro, indicating endocan could be a molecular target for pituitary adenoma treatment (34). Additionally, in human pituitary adenoma cells in vitro, silencing FGF2 inhibited cell proliferation and invasion capacity (35).…”

<p>LncRNA XIST depletion prevents cancer progression in invasive pituitary neuroendocrine tumor by inhibiting bFGF via upregulation of microRNA-424-5p</p>

“…From the invasive PitNET-related lncRNA expression dataset GSE51618, lncRNA XIST with high expression at a large FC in invasive PitNET was exhibited as the candidate lncRNAs (Figure 1A). In addition, a prior study revealed that siRNA-mediated bFGF gene silencing has the potential to inhibit the proliferation, migration and invasion of human PitNET cells 20. However, it is not clear whether XIST is involved in the regulation of bFGF in invasive PitNET.…”

The novel mechanism of anticancer effect on gastric cancer through inducing G0/G1 cell cycle arrest and caspase-dependent apoptosis in vitro and in vivo by methionine enkephalin

“…The cells in each group were homogenized using a homogenizer (POLYTRON PT2100; Kinematic, Luzern, Switzerland) with ice-cold lysis buffer containing 1 mM phenylmethylsulfonyl fluoride to extract total protein. The proteins were separated on 10% SDS-PAGE 26 and transferred to nitrocellulose membrane. After being blocked, the transferred proteins were incubated with relevant antibodies against OGFr (1:1,000; Sigma), Bax, BCL-2, caspase-3, PARP, β-actin (all above 1:1,000; Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.…”