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. 2021 Feb 26;14:1453–1463. doi: 10.2147/OTT.S281032

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© 2021 Yao et al.

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SPRR2B modulates MDM2-p53/p21 pathway and promotes gastric cancer cell cycle. (A) Protein level of SPRR2B was tested by immunoblotting in distinct cell lines, including nontumorous GES-1 cell line and gastric adenocarcinoma cell lines (AGS, MKN28, MKN45). (B) Knockdown assay was conducted by transfecting MKN45 with SPRR2B-siRNA, using scramble-siRNA as control. Rescue experiment was achieved by transfecting the cells with modified SPRR2B plasmids (pcDNA-SPRR2B°^pt). Immunoblotting results showed decreased expression of MDM2 and CDK1 in SPRR2B-knockdown cells while increased expression of p53 and p21. Re-expressing SPRR2B°^pt can abolish the effects of SPRR2B-siRNA. (C) Apoptosis assay by flow cytometry showed no statistically significant difference among three groups. (D) Flow cytometry tests confirmed the blockage effect of SPRR2B-siRNA on gastric cancer cell cycle, while transfecting SPRR2B°^pt (rescue group) help regain the cell cycle. (E) The downstream protein levels were tested after overexpressing SPRR2B in MKN45 cells. (F) Apoptosis assay indicated that SPRR2B has no significant effect on apoptosis. (G) Overexpressing SPRR2B facilitated cells enter G2/M cell cycle phase, while silencing MDM2 abolished this effect. (H) We hypothesized that SPRR2B promotes tumor cell proliferation perhaps through MDM2-p53/p21-CDK1 signaling pathway. Data was exhibited as mean ± SD. *Indicates P<0.05 by Student’s t-test.