Inhibiting the proliferation and metastasis of hilar cholangiocarcinoma cells by blocking the expression of vascular endothelial growth factor with small interfering RNA

Abstract

The aim of the present study was to investigate whether the proliferation and metastasis of hilar cholangiocarcinoma cells can be suppressed and whether apoptosis can be induced by small interfering RNA (siRNA) repression of vascular endothelial growth factor (VEGF). siRNA sequences targeting the gene were designed and the human hilar cholangiocarcinoma QBC939, HCCC-9810 and RBE cell lines were transfected with VEGF-siRNA plasmids for 48 h. Reverse transcription-quantitative polymerase chain reaction and western blotting measured the levels of VEGF-A, VEGF-C and matrix metalloproteinase 2 (MMP2) mRNA expression and protein content. The cell invasion potential was evaluated using the Transwell invasion and migration assay and the MTT assay was employed to detect the proliferation of hilar cholangiocarcinoma cells. Flow cytometry was used to quantify cell apoptosis and necrosis. Following the transfection of VEGF-siRNA, a significant reduction of mRNA and protein levels of VEGF-A, VEGF-C and MMP2 was observed in the hilar cholangiocarcinoma cells. The invasion, migration and proliferation of tumor cells were also notably decreased. The rate of tumor cell apoptosis was increased in the VEGF-siRNA group (15.42%) compared with the non-siRNA control (2.22%) and the negative control (2.71%) groups. It was concluded that blocking the expression of VEGF via VEGF-siRNA effectively inhibited the invasion, migration and proliferation, and induced apoptosis in hilar cholangiocarcinoma cells. These observations suggested that targeting VEGF with RNAi may be an effective therapeutic strategy for treating hilar cholangiocarcinoma.

Injured Endothelial Cell: A Risk Factor for Pulmonary Fibrosis

“…Jiao et al found that 27-Hydroxycholesterol (27HC) induced E(nd)MT in vascular ECs, and the process promoted EMT and migration in breast cancer cells through the alteration of the tumor microenvironment [93]. This was consistent with the studies that VEGF and syndecan-1 in the tumor stroma were significantly upregulated in dysfunctional ECs, simulated angiogenesis, and induced the migration and EMT of epithelial cells [94,95].…”

Role of Chemokines in the Biology of Cholangiocarcinoma

“…Tumors employ several mechanisms to establish a functional vascular system comprised of both blood and lymphatic vessels, to sustain cell growth [ 15 , 16 ]. CCA cells promote neo-vascularization by enhanced expression of angiogenic growth factors, whereas endothelial cells can release inflammatory chemokines to attract leukocytes and establish a pro-fibrotic and pro-angiogenic milieu, which in turn support migration, invasion and EMT [ 17 ]. The paracrine effects between CCA cells and surrounding stromal cells are summarized in Figure 1 .…”

Exploring the function of stromal cells in cholangiocarcinoma by three-dimensional bioprinting immune microenvironment model

“…A previous study shown that TEC induces tumor progression and invasion by activating a series of pathways ( 65 ). In addition, TEC can release inflammatory chemokines to attract leukocytes to establish a pro-fibrotic and pro-angiogenic microenvironment and promote migration, invasion, and EMT in CCA ( 66 ). Besides, CCA-associated endothelial cells highly express the erythropoietin receptor, which binds to cell-released erythropoietin in the tumor immune microenvironment, thus promoting proliferation, survival, and invasion of CCA cells ( 67 ).…”