| Objective: To study the molecular mechanism of Shuangshen Xiaoliu Decoction(SXD) inhibiting cell proliferation of gastric cancer and preventing the disease based on Wnt/β-catenin signaling pathway.Methods: Animal experiments were conducted using healthy male BALB/c-nu mice to establish a gastric cancer mouse model based on MNNG feeding method. According to different intervention methods, the mice were divided into model group, SXD high dose group, SXD low dose group, miR-338-3p agomir group, and combination group(SXD + miR-338-3p agomir). After 8 weeks of continuous intervention, the levels of serum motilin(MTL), pepsinogen I(PGI) gastrin(GAS), etc., were measured by ELISA; the pathological changes of tumor tissue were observed through HE staining; the mRNA expression levels of miR-338-3p, SIRT2, Wnt, and β-catenin in tumor tissue were detected by Real-time PCR; and the protein expression levels of SIRT2, Wnt, and β-catenin of tumor tissue were detected using Western blot technique.For cell experiment, according to different intervention methods, the mice were divided into model group, SXD high dose group, SXD low dose group, miR-338-3p mimics group, and miR-Con group. After the intervention, the cell activity was detected by MTT method, and the mRNA expression levels of cell miR-338-3p, SIRT2, Wnt, and β-catenin were detected by Real-time PCR method. Results: After intervention, compared with the model group, the serum MTL, PGI and GAS levels of mice in SXD high dose group, SXD low dose group, miR-338-3p agomir group and combination group were significantly increased(P < 0. 05). HE staining showed that compared with the model group, the gastric histopathology in SXD high dose group, SXD low dose group, miR-338-3p agomir group and combination group were improved in varying degrees. After intervention, compared with the model group, the mRNA expression level of miR-338-3p, SIRT2, Wnt, and β-catenin in tumor tissue of SXD high dose group, SXD low dose group, miR-338-3p agomir group, and the combination group were significantly increased(P < 0. 05). After the intervention, compared with the model group, the protein expression levels of SIRT2, Wnt, and β-catenin in tumor tissue of SXD high dose group, SXD low dose group, miR-338-3p agomir group, and combination group were significantly increased(P < 0. 05). After the intervention, compared with the model group, the cell survival rate of SXD high dose group, SXD low dose group, and miR-338-3p agomir group were significantly increased(P < 0. 05). Compared with the model group, the mRNA expression levels of miR-338-3p, SIRT2, Wnt, and β-catenin in gastric cancer cells in SXD high dose group, SXD low dose group, and miR-338-3p agomir group were significantly increased(P < 0. 05). Conclusion: The molecular mechanism of SXD to inhibit the proliferation of gastric cancer cells and prevent gastric cancer is related to the activation of Wnt/β-catenin signaling pathway through the targeted regulation of SIRT2 by miR-338-3p. |
