干扰HMGB1基因表达对食管鳞癌细胞侵袭迁移及放射敏感性的影响

doi:10.3969/j.issn.1004-616x.2021.05.002

Abstract

Abstract: OBJECTIVE: To examine the effect of HMGB1 silencing on proliferation, survival ability, migration and invasion of human esophageal squamous cell carcinoma after X-ray radiation. METHODS: Using siRNA technique, sequences of HMGB1 mRNA were synthesized and transfected into cultured EC9706 cells as the HMGB1 shRNA group. A negative sequence was synthesized and used as negative control (NC). Both groups were irradiated or not with Xrays. Protein levels of HMGB1 and metastasis-related molecules were determined using Western blot. MTS method, clone formation, and wound healing, Transwell assay were employed to examine the proliferation, survival ability, migration and invasion of the cells. Flow cytometry assay was used to analyze cell apoptosis rates. Western blot was used to examine the expression of cell. RESULTS: Western blot results demonstrated that silencing HMGB1 gene significantly reduced the HMGB1 protein expression compared with NC group (P < 0.01). MTS data demonstrated that,after irradiation, HMGB1 silencing significantly inhibited the proliferation of esophageal squamous cell carcinoma cells compared with NC group (P < 0.01). The data from the clone formation assay revealed that the radiosensitivity was increased after down-regulation of HMGB1 expression compared with NC group (P < 0.01). The apoptosis rate of tumor cells in HMGB1 shRNA group after irradiation was markedly increased (P < 0.01). Wound-healing assays show that Wound-healing rate of HMGB1 silencing cells was significantly lower than that of NC group[(20.78±4.38)% and (39.02 ±3.25)%, respectively, P < 0.01]. Transwell assays with matrigel show the invasion capability of HMGB1 silencing cells at 3.5 h was deeply suppressed compared with NC group (67.00±16.56 and 194.00±19.74, respectively, P < 0.01). Our data show that apoptosis rate of HMGB1 silencing cells was significantly higher than that of NC group[(20.67 ±1.38)% and (13.64 ±1.24)%, respectively, P < 0.01]. In addition, expressions of N-cadherin,Vimentin,MMP-2 and MMP-9 protein were downregulated,and expressions of Ecadherin were upregulated after HMGB1 silencing compared with NC group (P < 0.01). CONCLUSION: siRNA interference inhibited expression of the HMGB1 gene, reduced the proliferation, migration and invasion of cells,induced cell apoptosis,and increased radiosensitivity,followed by regulating the expression of metastasisrelated molecules in oesophageal squamous cell carcinoma cells.

Key words: esophageal squamous cell carcinoma, HMGB1, irradiation, migration, invasion, radiosensitivity

CLC Number:

R735.1

YANG Xingxiao, ZHANG Xueyuan, ZOU Naiyi, SHAN Bao'en, MA Ming, ZHU Shuchai. Effects of HMGB1 silencing on migration, invasion and radiosensitivity of esophageal squamous cell carcinoma cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(5): 327-333.

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Effects of HMGB1 silencing on migration, invasion and radiosensitivity of esophageal squamous cell carcinoma cells

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