| Objective To investigate the effect of shikonin on the biological function of nasopharyngeal carcinoma (NPC) cell lineCNE2 by regulating Hippo signaling pathway. Methods From January 2021 to December 2021, CNE2 cells in logarithmic growthphase were cultured by culture medium containing different concentrations of shikonin (0, 2.0, 4.0, 8.0, 16.0 mg/L) for 48 hours, thecell survival rate was detected by cell counting kit-8 (CCK-8) method. The apoptosis, colony formation, migration and invasion of CNE2cells were detected by flow cytometry, plate cloning, wound healing and Transwell assay. Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the relative expression of cell yes associated protein 1 (YAP1) and transcrip- tional coactivator with PDZ-binding motif (TAZ) with PDZ binding motif mRNA. The expression levels of YAP1, phosphorylated yes as- sociated protein 1 (p-YAP1), TAZ, phosphorylated transcriptional coactivator with PDZ-binding motif (p-TAZ), N-cadherin (N-cad), vi- mentin (Vim) and E-cadherin (E-cad) protein were detected by western blotting.Results Compared with 0 mg/L shikonin CNE2 cellsurvival rate (100%), colony formation number (514.67±25.81) cells, scratch healing rate (88.58±3.40)% , invasive cell number[(233.67±15.01) cells], YAP1 and TAZ mRNA and protein (1.01±0.02, 1.00±0.01, 0.68±0.04, 0.51±0.03), 2 mg/L shikonin (514.67±25.81) cells, scratch healing rate (88.58±3.40)% , invasive cell number [(233.67±15.01) cells], YAP1 and TAZ mRNA and protein(1.01±0.02, 1.00±0.01, 0.68±0.04, 0.51±0.03), 2 mg/L shikonin [(92.70±5.92)%, (452.33±22.72) cells, (69.91±3.03)%, (195.33±18.15)cells, (0.93±0.02), (0.91±0.05), (0.56±0.03), (0.44±0.02), 4 mg/L shikonin [(81.75±3.83)% , (308.33±22.12) cells, (53.61±3.21)% ,(153.33±10.02) cells , (0.81±0.03), (0.76±0.04), (0.45±0.03), (0.30±0.03), 8 mg/L shikonin [(54.93±3.89)% , (173.67±13.65) cells,(30.32±1.68)% , (92.67±6.66) cells, (0.65±0.03), (0.54±0.04), (0.31±0.03), (0.24±0.02), 16 mg/L shikonin [(33.89±2.14)% , (85.33±13.05) cells, (18.31±1.42)%, (52.33±6.03) cells, (0.41±0.02), (0.30±0.02), (0.18±0.02), (0.16±0.02) decreased (P<0.05), and the apopto- sis rate of CNE2 cells, the relative expression levels of p-YAP1, p-TAZ and E-cad protein increased with the increase of shikonin con- centration (P<0.05). The effects of shikonin were dose-dependent (P<0.05).Conclusion Shikonin can inhibit the malignant biologicalfunction of NPC cell line CNE2 cells, and its mechanism may be related to inhibiting the activation of the core downstream signaling fac-tors of the Hippo signaling pathway including YAP1 and TAZ proteins, and preventing epithelial mesenchymal transformation (EMT). |
