Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (25): 4025-4031.doi: 10.12307/2021.015

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Regulation of S100A4 gene silencing by ultrasound microbubbles on the stemness and epithelial-mesenchymal transformation of gastric cancer stem cells#br#

Xu Guilan1, Song Jiansheng2, Yang Shijiang2   

  1. 1Department of Ultrasonography, Wuhan Children’s Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430015, Hubei Province, China; 2Department of Emergency Surgery, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, Hubei Province, China 3Fujian Armed Police Corps Hospital, Fuzhou 350003, Fujian Province, China; 4Fujian Provincial Governmental Hospital, Fuzhou 350003, Fujian Province, China
  • Received:2020-07-01 Revised:2020-07-04 Accepted:2020-07-29 Online:2021-09-08 Published:2021-03-27
  • Contact: Yang Shijiang, MD, Associate chief physician, Department of Emergency Surgery, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, Hubei Province, China
  • About author:Xu Guilan, Attending physician, Department of Ultrasonography, Wuhan Children’s Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430015, Hubei Province, China Song Jiansheng, Master, Attending physician, Department of Emergency Surgery, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, Hubei Province, China

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Abstract: BACKGROUND: S100A4 is a member of S100 family of calcium binding proteins, which can bind to the corresponding target proteins, and then have an important impact on the proliferation and metastasis of tumor cells and pluripotent stem cells. Recent studies have shown that ultrasound targeted destruction of microbubbles can enhance gene transfection efficiency.
OBJECTIVE: To study the effect of S100A4 gene silencing by ultrasound targeted microbubble destruction on the stemness and epithelial-mesenchymal transformation of gastric cancer stem cells.
METHODS:  SGC-7901 human gastric cancer stem cells in logarithmic growth phase were harvested to isolate tumor stem cell spheres by serum-free suspension culture. The expression of EpCAM+CD44+ on the surface of tumor stem cells was detected by flow cytometry. The expression plasmid targeting S100A4 gene was constructed and transfected into EpCAM+CD44+ gastric cancer stem cells by ultrasound microbubble. At 48 hours after transfection, S100A4 expression was detected by western blot assay. The proliferation, clone formation, migration and invasion of gastric cancer stem cells were detected by CCK-8, clone formation test and Transwell cell. The expression of markers related to epithelial mesenchymal transformation was detected by western blot assay and qRT-PCR.
RESULTS AND CONCLUSION: Flow cytometry results showed that the proportion of EpCAM+CD44+ cells after enrichment of gastric cancer stem cells was significantly higher than that before enrichment (P < 0.05). The expression of S100A4 protein in the transfected group was significantly lower than that in the untransfected group (P < 0.01). At 48 hours after transfection, the proliferation rate, clone-forming ability, migration and invasion ability of gastric cancer stem cells significantly decreased, the expression of E-cadherin increased, and the expression of N-cadherin and Vimentin decreased. These findings indicate that RNA interference with S100A4 gene can inhibit the proliferation, clone-forming ability, migration and invasion of gastric cancer stem cells by regulating the expression of E-cadherin, N-cadherin and Vimentin.

Key words: stem cells, gastric cancer stem cells, ultrasound microbubbles, S100A4 gene, gene silencing, cell proliferation, cell migration, cell invasion, epithelial-mesenchymal transition

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