Figure 6.
Knocking down NORAD abated CAFs-mediated promotive effects in GC cells. CAFs were transfected with si-NORAD or IL-33 overexpression plasmids. The co-culture model of GC cells with CAFs was constructed to explore the interaction between GC cells and CAF. CAFs (1 × 105/well) were inoculated in the upper chambers, and AGC cells with NORAD knockdown were seeded in 24-well plates (below the chambers) at 5 × 105/well for 24 hours. a-c. RT-PCR evaluated the expression of NORAD, miR-496, and TGF-β1, Twist1, MMP-14 and VEGF-C in CAFs. d.CCK-8 tested the proliferation of AGS cells. E. TUNEL gauged unspecified cell death. f and g. Expression of apoptosis-associated proteins (Bak, Bax, bcl2, Caspase3 and Caspase9) in AGS cells was determined by WB. h-j. Transwell assay was utilized to test the migrative and invasive abilities of AGS cells. k and l. RT-PCR and ELISA were adopted to assess the mRNA and protein level of IL-33 in the supernatant.m. Expression of EMT-related markers (E-cadherin, ZO-1, Vimentin, N-cadherin, Snail, Slug, and MMP3) in AGS cells was evaluated by WB. All data were expressed as mean ± standard deviation. N = 3. **P < 0.01, ***P < 0.001(vs.Blank); &P < 0.05, &&P < 0.01, &&&P< 0.001(vs.CAFsh-NC); #P < 0.05, ##P < 0.01, ###P< 0.001(vs.CAFsh-NORAD)