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. 2023 Feb 21;19(5):1352–1368. doi: 10.7150/ijbs.79022

Figure 5.

Figure 5

Crosstalk with tumor cells enhances the affinity of neutrophils with tumor cells through interaction between integrins α6β1 and α6β4 with CD151. (A) RNA-seq analysis showed that several integrins, including α2, α3, α6, α10, αE, αv, β1, β4, β5, β6 and β8, were up-regulated in Edu-Neus. (B) The subunits α6 (ITGA6), β1 (ITGB1) and β4 (ITGB4) were the most significantly increased ones. (C) Co-culture with MKN45 cells up-regulated expression of the three subunits by qPCR assays, and treatment with Disitertide or LY-364947 could attenuate these effects. (D) Exogenous TGFβ1 increased the levels of these three subunits in neutrophils. (E) Prediction analysis of protein-protein interaction showed that ITGA6, ITGB1 and ITGB4 can interact with CD151. (F) Co-culture with neutrophils could increase CD151 expression in MKN45 cells. (G) FAM3C treatment could increase CD151 expression in MKN45 cells. (H) CD151 mRNA levels were correlated positively with FAM3C levels in gastric tumor tissues. (I) CD151 mRNA level in gastric tumor tissues was negatively associated with overall survival of gastric cancer patients. (J) Co-IP assays showed that ITGA6, ITGB1 and ITGB4 could bind CD151. (K) Subunit α6 was detected in neutrophils (red arrow, red) and CD151 was found in tumor cells (yellow arrow, yellow) using dual-color IHC assays for lymphatic cancer emboli. (*, P < 0.05; **, P < 0.01; ***, P < 0.001) (Edu-Neu, tumor-educated neutrophil).