Long noncoding RNAs HAND2-AS1 ultrasound microbubbles suppress hepatocellular carcinoma progression by regulating the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis

Figure 1 HAND2-AS1 expression in hepatocellular carcinoma development. A and B: HAND2-AS1 expression levels during hepatocellular carcinoma development were analyzed using bioinformatics techniques online (http://gepia.cancer-pku.cn/index.html; https://tnmplot.com/analysis/); C: HAND2-AS1 expression in tumor and surrounding tissues (35 pairings); D: The phrase represents HAND2-AS1. ^aP < 0.05.

Figure 2 HepG2 cell response to ultrasound microbubbles. Two hundred microliters of ultrasound microbubbles was added to HepG2 cells alone or incubated with si-HAND2-AS1. A: HAND2-AS1 expression; B: The proliferation of cells; C: Apoptosis of cells; D: Invasion of the cell; E: Proteins linked to epithelial-mesenchymal transition are expressed. ^aP < 0.05. UTMBs: Ultrasound microbubbles.

Figure 3 HAND2-AS1 directly targets the miRNA miR-873-5p. A: The target sites of HAND2-AS1 and miR-873-5p were identified (StarBase 3.0; https://starbase.sysu.edu.cn/); B: The HAND2-AS1 sequence; C: The relative luciferase activity value; D: RNA pull-down was employed to examine the strength of the binding; E: Cellular expression of miR-873-5p; F: Tissue expression of miR-873-5p (35 pairings); G: Expression of miR-873-5p. ^aP < 0.05.

Figure 4 The impact of miR-873-5p on HepG2 cells. A: Expression of miR-873-5p; B: HAND2-AS1; C: Cell proliferation; D: Apoptosis of cells Cell invasion; E: Expression of the snail protein, vimentin, and E-cadherin. ^aP < 0.05. UTMBs: Ultrasound microbubbles.

Figure 5 MiR-873-5p targets the 3'UTR of tissue inhibitor of matrix metalloproteinase-2. A: The 3’UTR of tissue inhibitor of matrix metalloproteinase-2 (TIMP2) targeted by miR-873-5p; B: The TIMP2 sequence; C: The relative luciferase activity value; D: Expression of miR-873-5p; E and F: TIMP2 mRNA and protein expression. ^aP < 0.05. TIMP2: Tissue inhibitor of matrix metalloproteinase-2.

Figure 6 Ultrasound microbubbles affect HepG2 cells via the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis. A and B: The expression of the proteins tissue inhibitor of matrix metalloproteinase-2, matrix metalloproteinase (MMP)-2, and MMP9; C: The proliferation of cells; D: Apoptosis of cells; E: Cell invasion; F: The expression of proteins linked to epithelial-mesenchymal transition. ^aP < 0.05. UTMBs: Ultrasound microbubbles; TIMP2: Tissue inhibitor of matrix metalloproteinase-2.

Figure 7 Ultrasound microbubbles inhibit tumor progression in vivo. The ultrasound microbubbles group (mice received injections of HAND2-AS1 microbubbles; 200 g; 1 MHz), and the control group (mice received injections of 0.9% NaCl alone; 1 MHz). A: Illustrations of tumors; B: Size of the tumor; C: Tumor mass; D: Expression of miR-873-5p and HAND2-AS1; E: Illustrations from immunohistochemistry analysis; F: Expression of proteins linked to epithelial-mesenchymal transition. ^aP < 0.05. UTMBs: Ultrasound microbubbles; TIMP2: Tissue inhibitor of matrix metalloproteinase-2; MMP: Matrix metalloproteinase.