AKR1B10 inhibits the proliferation and migration of gastric cancer via regulating epithelial-mesenchymal transition

Abstract

Gastric cancer (GC) is a common malignancy around the world with a poor prognosis. Aldo-keto reductase family 1 member B10 (AKR1B10) is indispensable to cancer development and progression, which has served as a diagnostic biomarker for tumors. In our study, we demonstrated that the expression of AKR1B10 in GC tissues was significantly lower compared with normal gastric tissues. Subgroup analysis showed that, according to the clinic-pathological factors, the effect of the AKR1B10 expression level on the prognosis of GC patients was significantly different. Moreover, reduced expression of AKR1B10 promoted the ability of GC cells in proliferation and migration. Furthermore, increased AKR1B10 levels resulted in the opposite trend in vitro. Moreover, AKR1B10 was correlated with epithelial-mesenchymal transition (EMT) in a significant way. In vivo experiment, knockdown of AKR1B10 promoted the growth of tumor, increased Vimentin, and E-cadherin significantly. In summary, AKR1B10 is considered as a tumor suppressor in GC and is a promising therapeutic target.

Keywords: AKR1B10; EMT; gastric cancer; migration; proliferation.

Figure 1

Expression of AKR1B10 in gastric cancer tissues. (A) Representative immunohistochemistry images showing in situ AKR1B10 expression in gastric cancer (GC) and normal tissues (scale bar = 100 μm). (B–D) IHC scores of AKR1B10 in (B) GC vs normal tissues, (C) tumors with and without lymph node invasion, and (D) TNM stage I–II vs III–IV. (E–G) overall survival analysis of (E) AKR1B10^pos vs AKRiB10^neg GC patients, and in subgroups overall survival analysis of TNM stage I–II (F) and III–IV (G). LNM, Lymph node metastasis.

Figure 2

Subgroup analysis of the influence of AKR1B10 expression on the survival of gastric cancer patients.

Figure 3

Nomograms to predict survival of gastric cancer patients. Points of each variable were obtained via a vertical line between each variable and the point scale. The predicted survival rate was correlated with the total points by drawing a vertical line from the total points scale to the overall survival.

Figure 4

Effect of AKR1B10 on the cell proliferation and migration ability of gastric cancer cells. (A) AKR1B10 expression in gastric cancer (GC) cell lines from the CCLE platform. (B) Relative expression of AKR1B10 in GC cell lines according to RNAseq results via the CCLE platform. (C–D) Immunoblot showing AKR1B10 protein levels in MKN45 cells transfected with AKR1B10-shRNA (C) and in AGS cells transfected with the AKR1B10 overexpression plasmid (D), and gray value analysis via ImageJ. (E–J) Proliferation rates (E–F), colony forming ability (G–H), and migration ability (I–J) of AKR1B10-KD and AKR1B10-OE GC cells. CCLE, Cancer Cell Line Encyclopedia. NC, negative control. KD, knockdown, AKR1B10-shRNA. VEC, vector. OE, overexpression, AKR1B10 overexpression plasmid. Data are presented as the mean ± SD (n = 3). ^*P < 0.05, ^**P < 0.01, ^***P < 0.001.

Figure 5

Correlation between AKR1B10 and epithelial-mesenchymal transition. (A) Correlation analysis of AKR1B10 and Vimentin gene expression levels in gastric cancer (GC) patients by the GEPIA platform. (B) Correlation analysis of AKR1B10 and E-cadherin gene expression levels in GC patients by the GEPIA platform. (C–E) Comparison of AKR1B10 (C), Vimentin (D) and E-cadherin (E) mRNA levels in 19 paired GC and normal tissues. (F) Correlation between AKR1B10 and Vimentin, and between AKR1B10 and E-cadherin mRNA levels in GC tissues. (G) MKN45 cells transfected with NC or KD and (H) AGS transfected with VEC or AKR1B10-OE. The bands were semi-quantified by ImageJ and the results are presented as the mean ± SD. VIM, Vimentin; ECAD, E-cadherin; TPM, transcripts per million. NC, negative control; KD, knockdown; VEC, vector; OE, overexpression. ^**P < 0.01, ^***P < 0.001.

Figure 6

AKR1B10 knockdown promotes gastric cancer tumor growth in vivo. (A–B) Total body weight (A) and tumor volume (B) of the mice. (C) Representative pictures of subcutaneous tumors harvested from NC and AKR1B10-KD group. (D) The weights of tumor masses. (E) The weights of mice without tumor masses. (F–I) Relative AKR1B10 (F), Vimentin (G) and E-cadherin (H) mRNA levels in tumors of the AKR1B10-KD or NC group, and their correlation (I). (J) Stratification of mice into cluster 1 and cluster 2 according to AKR1B10 mRNA levels, body weight and tumor weight. (K) Percentage of NC and AKR1B10-KD mice in each cluster. Data are presented as the mean ± SD. NC, negative control; KD, AKR1B10 knockdown. ^*P < 0.05, ^**P < 0.01.