Recombinant Newcastle disease virus rL-RVG enhances the apoptosis and inhibits the migration of A549 lung adenocarcinoma cells via regulating alpha 7 nicotinic acetylcholine receptors in vitro

Abstract

BackgroundThe aim of this study were to investigate the possible pro-apoptotic mechanisms of the recombinant Newcastle disease virus (NDV) strain rL-RVG, which expresses the rabies virus glycoprotein, in A549 lung adenocarcinoma cells via the regulation of alpha 7 nicotinic acetylcholine receptors (α7 nAChRs) and to analyze the relationships between α7 nAChR expression in lung cancer and the clinical pathological features.Methodsα7 nAChR expression in A549, LΑ795, and small-cell lung carcinoma (SCLC) cells, among others, was detected using reverse transcription polymerase chain reaction (RT-PCR). The optimal α7 nAChR antagonist and agonist concentrations for affecting A549 lung adenocarcinoma cells were detected using MTT assays. The α7 nAChR expression in A549 cells after various treatments was assessed by Western blot, immunofluorescence and RT-PCR analyses. Apoptosis in the various groups was also monitored by Western blot and TUNEL assays, followed by the detection of cell migration via transwell and scratch tests. Furthermore, α7 nAChR expression was examined by immunohistochemistry in lung cancer tissue samples from 130 patients and 40 pericancerous tissue samples, and the apoptotis in lung adenocarcinoma tissue was detected by Tunel assay, Then, the expression levels and clinicopathological characteristics were analyzed.ResultsOf the A549, LΑ795, SCLC and U251 cell lines, the A549 cells exhibited the highest α7 nAChR expression. The cells infected with rL-RVG exhibited high RVG gene and protein expression. The rL-RVG group exhibited weaker α7 nAChR expression compared with the methyllycaconitine citrate hydrate (MLA, an α7 nAChR antagonist) and NDV groups. At the same time, the MLA and rL-RVG treatments significantly inhibited proliferation and migration and promoted apoptosis in the lung cancer cells (P < 0.05). The expression of α7 nAChR was upregulated in lung cancer tissue compared with pericancerous tissue (P = 0.000) and was significantly related to smoking, clinical tumor-node-metastases stage, and histological differentiation (P < 0.05). The AI in lung adenocarcinoma tissue in high-medium differentiation group was lower than that in low differentiation group (p < 0.01).ConclusionsAn antagonist of α7 nAChR may be used as a molecular target for lung adenocarcinoma therapy. Recombinant NDV rL-RVG enhances the apoptosis and inhibits the migration of A549 lung adenocarcinoma cells by regulating α7 nAChR signaling pathways.Electronic supplementary materialThe online version of this article (10.1186/s12985-017-0852-z) contains supplementary material, which is available to authorized users.

Arachidonoylcholine and Other Unsaturated Long-Chain Acylcholines Are Endogenous Modulators of the Acetylcholine Signaling System

“…According to the literature data, the selective antagonist of nAChR methyllycaconitine at high concentrations (1 to 10 µM) inhibits the proliferation of the A549 cells. However, this effect is relatively small and manifests itself predominantly after 48 and 72 h of incubation [42]. Considering much lower affinity of acylcholines for nAChR, as compared with methyllycaconitine, their cytotoxicity might not arise solely from the nAChR inhibition.…”

Downregulation of RPN2 induces apoptosis and inhibits migration and invasion in colon carcinoma

“…The apoptosis level of SW1116 and SW480 cell transfection with RPN2 siRNA or control siRNA was assessed by flow cytometric assay. The overall cell apoptosis detection procedures were similar to the previously described procedures with slight modification on the cell numbers ( 23 ). In brief, the cells were washed and detached in 2 ml PBS with 2 mM ethylenediaminetetraacetic acid (EDTA) and centrifuged at 15,000 × g for 5 min.…”

Migration of gastric cancer is suppressed by recombinant Newcastle disease virus (rL-RVG) via regulating α7-nicotinic acetylcholine receptors/ERK- EMT

“…The rL-RVG, NDV, anti-RVG antibody and anti-NDV antibody were stored at − 80°C supplied by the Harbin Veterinary Research Institute (Harbin, China). The human gastric cancer cell lines SGC7901 and BGC were purchased from the Cancer Cell Repository (Shanghai Cell Bank,2016- [11][12][13][14][15][16][17][18][19][20] and the American Type Culture Collection (Manassas,VA,USA). The cell line has been authenticated by STR (short tandem repeats, STR) and has been tested for mycoplasma contamination (Additional file 1).…”

“…Alpha 7-nAChR may be a potential therapeutic key point for the treatment of stomach cancer. Previous studies have also demonstrated that the recombinant avirulent NDV LaSota strain, expressing the rabies virus glycoprotein (rL-RVG), could induce the apoptosis of gastric cancer cells as well as suppress the migration of lung cancer cells by regulating α7-nAChR [13,14]. However, it still remains unclear whether rL-RVG could also suppress the migration of gastric cancer and its underlying mechanisms.…”