Knockdown of RAI14 suppresses the progression of gastric cancer

Figures & data

Figure 1 RAI14 downregulation is associated with prognosis of gastric cancer patients and reduces proliferation and viability of gastric cancer cells.

Notes: (A) The expression of RAI14 mRNA in gastric cancer tissues (red box) and normal tissues (black box) from GEPIA. (B) The survival curve of gastric cancer patients with different level of RAI14 mRNA from GEPIA. (C) The survival curve of gastric cancer patients from KM plotter analysis. (D) The expression of RAI14 mRNA in MNK45 and AGS cells by RT-PCR assay. (E, F) Expression of RAI14 protein in MKN45 (E) and AGS (F) cells with or without siRNA-RAI14. (G, H) Cell proliferation curve of MKN45 (G) and AGS (H) cells with or without siRNA-RAI14 from CCK8 assay. (I, J) Colony formation ability of MKN45 (I) and AGS (J) cells with or without siRNA-RAI14. Blank: cells without any treatment; NC: negative control, cells transfected with siRNA negative control; RAI14-KD: cells transfected with siRNA-RAI14, RAI14 knockdown. *P<0.05. Results are from three repeated experiments.
Abbreviations: GEPIA, gene expression profiling interactive analysis; RT-PCR, reverse-transcription PCR; STAD, stomach adenocarcinoma.

Figure 2 RAI14 knockdown inhibits cell migration and invasion, promotes apoptosis of gastric cancer cells.

Notes: (A, B) Cell migration of MKN45 (A) and AGS (B) cells with or without siRNA-RAI14 was assessed using Transwell assay. (C, D) Cell migration of MKN45 (C) and AGS (D) cells with or without siRNA-RAI14 was assessed. (E) Cell apoptosis of MKN45 (left) and AGS (right) cells with or without siRNA-RAI14 was evaluated using flow cytometry assay. (F) Expression of apoptosis-related proteins (Bcl-2, Bax, active Caspase3 and 9) in MKN45 and AGS cells with or without siRNA-RAI14. *P<0.05, **P<0.01. Results are from three repeated experiments.
Abbreviations: KD, knockdown; NC, normal control.

Figure 3 RAI14 knockdown inhibits the activation of Akt pathway.

Notes: (A) Western blot results of key proteins in Akt pathway in MKN45 cell with or without siRNA-RAI14. (B) Expression of p-Akt in MKN45 cell with siRNA-RAI14, IGF-1 or LY294002. (C) Cell proliferation curve of MKN45 cells with siRNA-RAI14, IGF-1 or LY294002 from CCK8 assay. Results are from three repeated experiments. IGF-1: cells were treated with IGF-1 (50 ng/mL). *P<0.05, **P<0.01.
Abbreviations: KD, knockdown; NC, normal control.

Figure 4 RAB31 is a potential target gene for RAI14.

Notes: (A) The correlation analysis from GEPIA between expression of RAI14 mRNA and RAB31 mRNA in gastric cancer. (B) Expression of RAB31 mRNA in MKN45 cells with or without siRNA-RAI14. (C) Expression of RAB31 mRNA in MKN45 cells with or without RAB31-overexpression vector. RAB31-OV: cells were transfected with RAB31-overexpression vector. (D) Expression of RAI14, RAB31 and p-Akt proteins in MKN45 cells by Western blot assay. (E) Cell proliferation curve of MKN45 cells with or without RAB31-overexpression vector from CCK8 assay. Results are from three repeated experiments. *P<0.05, **P<0.01.
Abbreviations: GEPIA, gene expression profiling interactive analysis; KD, knockdown; NC, normal control; OV, overexpression.

Figure S1 RAI14 overexpression promotes cell migration and invasion, reduces cell apoptosis in gastric cancer cells.

Notes: (A) Cell migration of MKN45 (left) and AGS (right) cells with RAI14 overexpression vector was assessed using Transwell assay. (B) Cell migration of MKN45 (left) and AGS (right) cells with RAI14 overexpression vector was assessed. (C) Cell apoptosis of MKN45 (left) and AGS (right) cells with RAI14 overexpression vector was evaluated using flow cytometry assay. (D) Expression of apoptosis-related proteins (Bcl-2, Bax, active Caspase3 and 9) in MKN45 and AGS cells with RAI14 overexpression vector. *P<0.05. Results are from three repeated experiments.
Abbreviations: KD, knockdown; NC, normal control; OV, overexpression.