LncRNA H19 induced by helicobacter pylori infection promotes gastric cancer cell growth via enhancing NF-κB-induced inflammation

Abstract

Background: The aim of this study was to investigate the role of long non-coding RNA (lncRNA) H19 in gastric cancer (GC) with Helicobacter pylori (H. pylori).

Methods: H19 expression in peripheral blood from H. pylori+/- GC patients and healthy donors (control) as well as in GC tissues and cells were detected by qRT-PCR. Cell proliferation was evaluated by CCK-8 assay. Cell migration and invasion were evaluated by Transwell assay. The levels of pro-inflammatory cytokines were determined by ELISA. The protein levels of IκBα, p-IκBα and p65 were determined by western blotting.

Results: H19 expression was upregulated in H. pylori-infected GC tissues and cells. Furthermore, H. pylori promoted GC cell viability, migration, invasion and inflammatory response. Moreover, H19 overexpression promoted the proliferation, migration and invasion of H. pylori-infected GC cells via enhancing NF-κB-induced inflammation.

Conclusions: LncRNA H19 promotes H. pylori-induced GC cell growth via enhancing NF-κB-induced inflammation.

Keywords: Gastric cancer; H. pylori; H19; NF-κB signaling pathway.

Fig. 1

H19 was upregulated in HP-infected GC tissues and cells. a: H19 expression in sera from HP +/− GC patients and healthy donors (control) was detected by qRT-PCR. b: H19 expression in gastric carcinoma and adjacent normal tissues from HP +/− GC patients was detected by qRT-PCR. N = 30 for each group. c: H19 expression in SGC-7901 cells and gastric mucosa epithelial GES-1 cells following 24 h of HP infection at MOI (=0, 1, 10, 100) was detected by qRT-PCR. D: H19 expression in SGC-7901 and GES-1 cells following HP infection at MOI = 100 for 0, 6, 12, 24 h was detected by qRT-PCR. Each experiment was repeated three times. *p < 0.05 vs. control or adjacent tissue or MOI =0 or 0 h; #p < 0.05 vs. HP-

Fig. 2

The effect of HP infection on GC cell behaviors. SGC-7901 cells were infected with HP at MOI = 100 for 24 h. a: Cell viability was analyzed using a CCK-8 kit. b, c: Cell migration and invasion were analyzed using transwell assays. d: The levels of pro-inflammatory cytokines were determined by ELISA. Each experiment was repeated three times. *p < 0.05 vs. control

Fig. 3

The effect of H19 on cell behaviors of SGC-7901 cells infected with HP. SGC-7901 cells were transfected with H19, si-H19 and their respective controls, followed by HP infection at MOI = 100 for 24 h. a The overexpression and knockdown efficiencies of H19 were confirmed by qRT-PCR. b Cell proliferation was analyzed using a CCK-8 kit. c Cell migration and invasion were analyzed using transwell assays. Each experiment was repeated three times. *p < 0.05 vs. vector or control; #p < 0.05 vs. si-ctrl or HP+ vector;& p < 0.05 vs. HP + si-ctrl

Fig. 4

H19 promoted HP-induced GC cell proliferation, migration, and invasion via NF-κB signaling pathway. a: The protein levels of phosphorylated (p)-IκBα, total (t)-IκBα, and nuclear p65 in SGC-7901 cells infected with HP at MOI = 100 for 24 h were determined by western blotting. b: The effect of H19 on HP-activated NF-κB signaling pathway was determined by western blotting. c-e: SGC-7901 cells were transfected with H19 overexpression vector and corresponding vector controls, followed by treatment with BAY11–7082 and PDTC. After which, cells were infected with HP at MOI = 100 for 24 h. c: Cell proliferation was analyzed using a CCK-8 kit. d: Cell migration and invasion were analyzed using transwell assays. e: The pro-inflammatory cytokines levels were determined by ELISA. Each experiment was repeated three times. *p < 0.05 vs. control or HP + vector; #p < 0.05 vs. HP + H19+ DMSO

Fig. 5

The reversal effect of NF-κB inhibitors on H19 effect was dependent on HP infection. SGC-7901 cells were transfected with H19 overexpression vector and corresponding vector controls, followed by treatment with BAY11–7082 and PDTC. a Cell proliferation was analyzed using a CCK-8 kit. b Cell migration and invasion were analyzed using transwell assays. c The pro-inflammatory cytokines levels were determined by ELISA. Each experiment was repeated three times. *p < 0.05 vs. vector