Osteopontin promotes gastric cancer progression via phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway

Figure 2 Osteopontin knockdown inhibits proliferation, invasion and metastasis of gastric cancer cells. A: Fluorescence microscope observation transfection efficiency of SGC-7901 cells after being transfected with osteopontin-short hairpin RNA (shRNA) (× 100); B: 3distinct, sequence-specific OPN shRNAs (OPN-shRNA1; OPN-shRNA2; OPN-shRNA3) and negative control shRNA were designed, and the OPN-shRNA3 has the best interference efficiency of OPN; C: Western blot assay of OPN protein levels in SGC-7901 cells 48  h after transfection; D: Densitometry analysis of the protein bands of OPN proteins in SGC-7901 cells 48  h after they were transfected; E: 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide assay observation the relative proliferation rate at 0, 24, 48 and 96 h in SGC-7901 cells following transfection, compared with control group; F: Microscope observation effect of the invasive ability of OPN knockdown in SGC-7901 cells was assessed by the Transwell matrigel-coated assay (× 100); G: Microscope observation of the migrative ability of OPN knockdown on SGC-7901 cells was assessed by the Transwell assay (× 100); H and I: Cells invading and migrating through the membrane were counted in 3 random fields for respective group. Data are shown as the means ± SE (n = 3). ^aP < 0.05; ^bP < 0.01; ^cP < 0.001. OPN: Osteopontin; shRNA: Short hairpin RNA; NC-shRNA: Negative control shRNA.