Ultrasound-Targeted Microbubble Destruction Enhances the Inhibitive Efficacy of miR-21 Silencing in HeLa Cells
Author(s) -
Shengli Zhao,
Jing Xie,
Changhua Zhao,
Wen Cao,
Yangping Yu
Publication year - 2021
Publication title -
medical science monitor
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.636
H-Index - 85
eISSN - 1643-3750
pISSN - 1234-1010
DOI - 10.12659/msm.923660
Subject(s) - transfection , hela , flow cytometry , apoptosis , gene silencing , cell growth , microbiology and biotechnology , pi3k/akt/mtor pathway , cancer research , cell culture , protein kinase b , wnt signaling pathway , cell , western blot , chemistry , biology , signal transduction , gene , genetics , biochemistry
Background Previous studies have shown that miR-21 upregulation is related to the aggressive development of cervical cancer. Ultrasound-targeted microbubble destruction (UTMD) is a method that increases the absorption of targeted genes or drugs by cells. We focus on the role of UTMD-mediated miR-21 transfection in HeLa cells, a cervical cancer cell line. Material/Methods The effects of different ultrasound intensities on the transfection efficiency of miR-21-enhanced green fluorescent protein (EGFP) and miR-21 inhibitor-EGFP plasmids were determined by flow cytometry. The effects of UTMD-mediated miR-21 transfection on HeLa cell proliferation, apoptosis, migration, and invasion were measured by CCK-8, flow cytometry, wound healing experiments, and transwell migration assay, respectively. Western blot and real-time quantitative PCR were used to detect the expression of tumor-related genes. Results When the ultrasound intensity was 1.5 W/cm2, the miR-21 plasmid had the highest transfection efficiency. Exogenous miR-21 promoted cell proliferation, migration, and invasion, and inhibited cell apoptosis in HeLa cells. Treatment of cells with UTMD further enhanced the effects of miR-21-EGFP and miR-21 inhibitor-EGFP. In addition, miR-21 overexpression significantly increased the expression of p-Akt, Akt, Bcl-2, Wnt, β-catenin, matrix metalloprotein-9 (MMP-9), and epidermal growth factor (EGFR) levels, and decreased Bax expression. The regulatory role of miR-21 inhibitor-EGFP was opposite to that of miR-21-EGFP. After UTMD, miR-21-EGFP and miR-21 inhibitor-EGFP had more significant regulatory effects on these genes. Conclusions Our research revealed that an ultrasound intensity of 1.5 W/cm2 is the best parameter for miR-21 transfection. UTMD can enhance the biological function of miR-21 in HeLa cells, and alter the effect of miR-21 on apoptosis, metastasis, and phosphorylation genes.
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