Abstract:Objective: To investigate the effect of circ-0086414 targeting miR-155-5p on the inhibition of Hippo/Yap signaling pathway in regulating the proliferation, migration and invasion of oral squamous cell carcinoma cells. Methods: RT-PCR was used to detect the expression of circ-0086414 in HIOEC, Tca8113, CAL27 and SCC-9 cells.SCC-9 cells which were not transfected with any plasmid were set as control group; SCC-9 cells transfected with pcDNA-circ-0086414, pcDNA-NC, miR-155-5p inhibitor and miR-NC inhibitor plasmids were set as pcDNA-circ-0086414 group, pcDNA-NC group, miR-155-5p group and miR-NC group respectively; the pcDNA-circ-0086414 plasmid was transfected into SCC-9 cells, and 5 μmol/L Verteporfin was co-cultured and set as Verteporfin group. The expression of circ-0086414 and miR-155-5p was detected by RT-PCR; CCK-8 was used to detect cell proliferation; Transwell chamber method was used to detect the invasive ability of cells; the cell migration ability was detected by scratch test; the expression of Lats1, p-Lats1, YAP, p-YAP protein in cells was detected by Western blot; the target relationship between circ-0,086,414 and miR-155-5p was verified by double luciferase reporting system experiment. Results: Compared with human immortalized oral epithelial cells HIOECcirc-0086414 (1.00±0.10) and miR-155-5p (1.00±0.09), the expression of circ-0086414 (0.73±0.07), (0.51±0.05) and (0.36±0.03) in oral squamous cell carcinoma cell lines Tca8113, CAL27 and SCC-9 significantly decreased, and the expression of miR-155-5p (1.36±0.11), (1.57±0.13) and (1.89±0.15) significantly increased (P<0.05); the expression of circ-0086414 and miR-155-5p in SCC-9 cells changed most significantly (P<0.05). Compared with the control group, the expression of circ-0086414 in pcDNA-circ-0086414 group was significantly higher (1.26±0.12), and the expression of miR-155-5p in miR-155-5p group was significantly lower (0.73±0.07) (P<0.001). Compared with the control group, the proliferation, invasion (123.56±5.87) and migration (16.90±1.62) of cells in pcDNA-circ-0086414 group significantly decreased (P<0.05). Compared with pcDNA-circ-0086414 group, the proliferation, invasion and migration of cells in Verteporfin group significantly increased (P<0.05). Compared with the control group, the expression of Last1 (1.12±0.10), p-Last1 (1.49±0.12) and p-YAP (1.4±0.11) protein in pcDNA-circ-0086414 group significantly increased, while the expression of YAP protein (0.45±0.04) significantly decreased (P<0.001). Compared with pcDNA-circ-0086414 group, the expression of Last1 (0.61±0.06), p-Last1 (0.82±0.08) and p-YAP protein (0.58±0.05) in the cells of Verteporfin group significantly decreased, and the expression of YAP protein (0.93±0.09) significantly increased (P<0.001). The luciferase activity of wild-type circ-0086414 (circ-0086414-WT) miR-155-5p group (0.26±0.03) was significantly lower than that of miR-NC group (P<0.05). Conclusion: Overexpression of circ-0086414 can inhibit the proliferation, invasion and migration of oral squamous cell carcinoma SCC-9 cells. Its mechanism is related to inhibition of miR-155-5p expression and activation of Hippo Yap signal pathway.
[1] Russo D,Mariani P,Caponio VCA,et al.Development and validation of prognostic models for oral squamous cell carcinoma:a systematic review and appraisal of the literature[J].Cancers,2021,13(22):5755. [2] Ling Z,Cheng B,Tao X.Epithelial-to-mesenchymal transition in oral squamous cell carcinoma:challenges and opportunities[J].International Journal of Cancer,2021,148(7):1548-1561. [3] Lin NC,Hsien SI,Hsu JT,et al.Impact on patients with oral squamous cell carcinoma in different anatomical subsites:a single-center study in Taiwan[J].Scientific Reports,2021,11(1):1-9. [4] Zhao W,Cui Y,Liu L,et al.Splicing factor derived circular RNA circUHRF1 accelerates oral squamous cell carcinoma tumorigenesis via feedback loop[J].Cell Death & Differentiation,2020,27(3):919-933. [5] Su W,Wang Y,Wang F,et al.Circular RNA hsa_circ_0007059 indicates prognosis and influences malignant behavior via AKT/mTOR in oral squamous cell carcinoma[J].Journal of Cellular Physiology,2019,234(9):15156-15166. [6] Gao F,Han J,Wang Y,et al.Circ_0109291 promotes cisplatin resistance of oral squamous cell carcinoma by sponging miR-188-3p to increase ABCB1 expression[J].Cancer Biotherapy & Radiopharmaceuticals,2022,37(4):233-245. [7] Fu S,Chen HH,Cheng P,et al.MiR-155 regulates oral squamous cell carcinoma Tca8113 cell proliferation,cycle and apoptosis via regulating p27Kip1[J].Eur Rev Med Pharmacol Sci,2017,21(5):937-944. [8] Li X,Wang S,Mu W,et al.Reactive oxygen species reprogram macrophages to suppress antitumor immune response through the exosomal miR-155-5p/PD-L1 pathway[J].Journal of Experimental & Clinical Cancer Research,2022,41(1):1-19. [9] Lee NH,Kim SJ,Hyun J.MicroRNAs regulating hippo-YAP signaling in liver cancer[J].Biomedicines,2021,9(4):347. [10] Yin F,Dong J,Kang LI,et al.Hippo-YAP signaling in digestive system tumors[J].American Journal of Cancer Research,2021,11(6):2495. [11] Zheng DW,Deng WW,Song WF,et al.Biomaterial-mediated modulation of oral microbiota synergizes with PD-1 blockade in mice with oral squamous cell carcinoma[J].Nature Biomedical Engineering,2022,6(1):32-43. [12] Zhang Q,Wang W,Zhou Q,et al.Roles of circRNAs in the tumour microenvironment[J].Molecular Cancer,2020,19(1):1-16. [13] Yang X,Ye T,Liu H,et al.Expression profiles,biological functions and clinical significance of circRNAs in bladder cancer[J].Molecular Cancer,2021,20(1):1-25. [14] 苏文,王宇帆,王锋,等.环状RNA hsa_circ_0002203对口腔鳞状细胞癌细胞增殖,迁移,侵袭和凋亡能力的影响[J].华西口腔医学杂志,2019,37(5):509-515. [15] 房旭,刘维贤.环状RNA hsa_circ_0009043在口腔鳞状细胞癌中的表达及临床意义[J].中国医科大学学报,2022,51(7):633-637.