The miRNA, miR-125b, Inhibited Invasion and Metastasis of Gastric-Cancer Cells by Triggering the STAT3 Signaling Pathway

Xiangqian Xu; Zhongqin Dang; Junping Zhang; Yingpu Feng; Zheng Wei

doi:10.2147/CMAR.S259513

The miRNA, miR-125b, Inhibited Invasion and Metastasis of Gastric-Cancer Cells by Triggering the STAT3 Signaling Pathway

Cancer Manag Res. 2020 Sep 17:12:8569-8580. doi: 10.2147/CMAR.S259513. eCollection 2020.

Authors

Xiangqian Xu^#^{1

2}, Zhongqin Dang^#², Junping Zhang³, Yingpu Feng⁴, Zheng Wei^{1

3}

Affiliations

¹ Gastroenterology Department, Henan University of Chinese Medicine, Zhengzhou, Henan, 450000, People's Republic of China.
² Gastroenterology Department, The Second Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan, 450002, People's Republic of China.
³ Oncology Department, Henan Academy Institute of Traditional Chinese Medicine, Zhengzhou, Henan, 450000, People's Republic of China.
⁴ Oncology Department, Henan Province People's Hospital, Zhengzhou, Henan, 450003, People's Republic of China.

^# Contributed equally.

Abstract

Objective: To investigate the function and the mechanism of miR-125b in the invasion and metastasis of gastric cancer and provide experimental basis for finding and developing new therapeutic strategies for gastric cancer.

Methods: The difference of miR-125b expression in gastric cancer tissues and adjacent tissues was detected by qRT-PCR. The same test was performed in different gastric cancer cell lines. The effect of miR-125b on SGC-7901 and BGC-823 gastric cancer cell viability was examined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Transwell assay was used to detect the effect of miR-125b on invasion and metastasis of gastric cancer cells. The target gene STAT3 of miR-125b was identified and validated by dual luciferase reporter assay. Western blot assay and immunofluorescence staining were used to detect the effect of miR-125b on the expression and distribution of STAT3 protein. The inhibitor and activator of STAT3 were used to confirm the effect of STAT3 on invasion and metastasis of gastric cancer cells. Peritoneal metastasis experiment and IHC were used to study the inhibitory effect of miR-125b on the metastasis of gastric cancer in vivo.

Results: The results of qRT-PCR showed that 125b expression was significantly lower in gastric cancer than in adjacent tissues, which indicated poor prognosis for gastric-cancer patients. Furthermore, two gastric-cancer cell lines, SGC-7901 and BGC-823, exhibited lower miR-125b levels than the normal cell line HEK293. After treatment with miR-125b mimics, cell proliferation was markedly inhibited. Meanwhile, the invasion and metastasis of gastric cancer cells were also inhibited after treated with miR-125b mimics. We also identified the signal transducer and activator of transcription 3 (STAT3) as a potential target of miR-125b based on patient data from The Cancer Genome Atlas (TCGA). Dual luciferase assays revealed that miR-125b directly inhibited STAT3 by binding to its 3'-untranslated region (UTR). Immunofluorescence assay showed that miR-125b could affect the subcellular distribution of STAT3. Moreover, treatment with miR-125b mimics or stattic inhibited invasion and migration in the gastric cancer cell lines, and IL-6 could reverse the inhibitory effect. Finally, nude mice xenografted with gastric-cancer cells expressing miR-125b mimics exhibited smaller tumors and lower transfer rates than mice engrafted with control group cells.

Conclusion: These data suggested that miR-125b inhibited invasion and metastasis in gastric cancer by inhibiting STAT3; therefore, miR-125b and STAT3 could be potential therapeutic targets in the treatment of gastric cancer.

Keywords: STAT3; gastric cancer; invasion and metastasis; miR-125b.

Grants and funding

This study was financially supported by grants from the National Natural Science Foundation of China (Project No. 81873290), the Henan Traditional Chinese Medicine (TCM) Foundation (No. 2017ZY1009, 2019ZYBJ28) and the Scientific and Technological Project of Henan Province, China (No. 182102311164). Xiangqian Xu and Zhongqin Dang contributed equally to this work and should be considered as co-first authors.