Circular RNA SMARCA5 Modulates Epithelial-Mesenchymal Transformation, Proliferation, and Metastasis of Nasopharyngeal Carcinoma Cells via microRNA-582-3p/Phosphatase and Tensin Homolog Axis

Abstract

The action mechanism in which circular RNA (circ) SMARCA5 targeted nasopharyngeal carcinoma (NPC) cell proliferation, migration, invasion, and apoptosis via microRNA (miR)-582-3p/phosphatase and tensin homolog (PTEN) axis was explored. The examination was performed via reverse transcription-quantitative polymerase chain reaction (RT-qPCR), discovering that circSMARCA5 was elevated while miR-582-3p was silenced in NPC tissues and cells. E-cadherin and N-cadherin were detected. The results illustrated transfection with si-circSMARCA5 or miR-582-3p-mimic was available to repress cancer cell advancement, and E-cadherin was augmented. Transfection with pcDNA 3.1-circSMARCA5 or miR-582-3p-inhibitor was available to accelerate cancer cell advancement, and N-cadherin was augmented. MiR-582-3p-inhibitor blocked the suppression of si-circSMARCA5 on NPC. The si-PTEN blocked the malignant behavior of pcDNA 3.1-circSMARCA5 against NPC. The binding sites between circSMARCA5 and miR-582-3p and between miR-582-3p and PTEN were verified. Linear analysis results illuminated the expression pattern of circSMARCA5 was opposite to miR-582-3p, while the expression pattern of circSMARCA5 was positively associated with PTEN. In brief, the results of the research clarified circSMARCA5 modulated NPC cells' vital movement via the miR-582-3p/PTEN molecular axis.

Figure 1

CircSMARCA5 is elevated in NPC tissues and cells. (a) RT-qPCR test of circSMARCA5 in NPC tissues and adjacent normal tissues; (b) RT-qPCR examination of circSMARCA5 in human embryonic nasopharyngeal epithelial cell HENE and NPC cell lines CNE1, HONE1, and CNE2. ^∗P < 0.05.

Figure 2

Silenced circSMARCA5 restrains CNE2 cell advancement. (a) RT-qPCR examination of circSMARCA5 after transfection with si-circSMARCA5; (b) MTT assay test of the impact of silenced circSMARCA5 on CNE2 cell proliferation; (c) cell colony formation assay examination of CNE2 cell proliferation after silencing circSMARCA5; (d, e) Transwell assay test of CNE2 cell migration and invasion after silencing circSMARCA5; (f) western blot examination of E-cadherin and N-cadherin after silencing circSMARCA5; (g) flow cytometry test of CNE2 cell apoptosis rate after silencing circSMARCA5. All experiments were repeated three times, ^∗P < 0.05.

Figure 3

CircSMARCA5 targets miR-582-3p. (a) Through https://starbase.sysu.edu.cn/ forecast of the combination domain of miR-582-3p and circSMARCA5; (b) luciferase activity assay verification of the targeting of circSMARCA5 with miR-582-3p; (c) RT-qPCR detection of miR-370-3p after silencing circSMARCA5; (d, e) RT-qPCR examination of miR-582-3p in NPC tissues and cell lines. (f) circSMARCA5 was negatively associated with miR-582-3p. All experiments were repeated three times, ^∗P < 0.05.

Figure 4

Elevated miR-582-3p represses CNE2 cell advancement. (a) After transfection of miR-582-3p-mimic into CNE2 cells, RT-qPCR test of miR-582-3p; (b) MTT assay examination of cell proliferation; (c) cell colony formation assay test of cell proliferation; (d, e) Transwell assay test of cell migration and invasion ability; (f) western blot examination of E-cadherin and N-cadherin; (g) flow cytometry test of cell apoptosis. All experiments were repeated three times, ^∗P < 0.05.

Figure 5

Suppressive miR-582-3p turns around the repression of silenced circSMARCA5 on NPC cells (a) Co-transfection of si-circSMARCA5 and miR-582-3p-inhibitor was into CNE2 cells, and RT-qPCR test of miR-582-3p; (b) MTT assay examination of cell proliferation; (c) cell colony formation assay test of cell proliferation; (d, e) Transwell assay examination of cell migration and invasion; (f) western blot test of E-cadherin and N-cadherin; (g) flow cytometry examination of cell apoptosis. All experiments were repeated three times, ^∗P < 0.05.

Figure 6

MiR-582-3p targets PTEN. (a) The binding domain of miR-582-3p with PTEN; (b) luciferase activity assay verification of the targeting of miR-582-3p and PTEN; (c, d) RT-qPCR and western blot detection of targeted regulation of miR-582-3p with PTEN; (e, f) PTEN in NPC tissues and cell lines. All experiments were repeated three times, ^∗P < 0.05.

Figure 7

CircSMARCA5 elevates NPCS via modulating miR-582-3p/PTEN axis. (a, b) After co-transfection of si-PTEN and pcDNA 3.1-circSMARCA5 into CNE2 cells, RT-qPCR and western blot examination of PTEN; (c) MTT assay test of cell proliferation; (d) cell colony formation assay examination of cell proliferation; (e, f) Transwell assay test of cell migration and invasion; (g) western blot test of E-cadherin and N-cadherin; (h) fow cytometry examination of cell apoptosis; (i) linear calculation analysis of the relevance of PTEN with circS MARCA5. All experiments were repeated three times, ^∗P < 0.05.