HMGB1 siRNA干扰对食管鳞癌细胞放射敏感性的影响

doi:10.3969/j.issn.1004-616x.2020.04.004

Abstract

Abstract: OBJECTIVE: To examine the effect of HMGB1 silencing by siRNA on cell proliferation, survival ability, and cell apoptosis of human esophageal squamous cell carcinoma, KYSE30, after X-ray radiation. METHODS: There were three study groups. The HMGB1 siRNA group:siRNA based on the sequences of the HMGB1 mRNA were synthesized and were transfected into the cultured KYSE30 cells. The negative control group:a negative siRNA was synthesized and transfected. The blank control group:no transfection was conducted. Expression of HMGB1 at the mRNA and protein levels was determined using quantitative real-time PCR (qPCR) and Western blot, respectively. The effect of HMGB1 knockdown on the proliferation,survival ability,cell apoptosis rate and the expression of cell apoptosis-related proteins of tumor cells were examined by MTS, clonal formation, flow cytometry and Western blot assays, respectively. RESULTS: Both mRNA and protein expression of HMGB1 were significantly reduced after silencing the HMGB1 gene (P < 0.01). MTS data demonstrated that HMGB1 knockdown,followed by irradiation,significantly inhibited the proliferation of KYSE30 cells compared with the negative control and the blank control groups (P < 0.05). Data from the clonal formation assay revealed that the values of D₀ were 2.57 Gy,2.54 Gy,1.55 Gy; the values of D_q were 1.69 Gy,1.65 Gy,1.30 Gy;the values of SF₂ were 0.27,0.27,0.13;the values of N were 1.93,1.91,2.31 in the blank control,negative control and HMGB1 siRNA groups,respectively (all P < 0.01). The apoptosis rate of tumor cells in the HMGB1 siRNA group after irradiation was markedly increased, followed by the downregulation of bcl-2 expression and the upregulation of bax, caspase-3 expression compared with negative control and blank control groups (P < 0.01). CONCLUSION: SiRNA interference technology inhibited the expression of HMGB1 gene in KYSE30 cells,reduced the proliferation and viability ability of cells, induced cell apoptosis, and increased radiosensitivity after X-ray exposure, which may be associated with regulating the expression of bcl-2,bax and caspase-3.

Key words: high mobility group box 1, siRNA interference, irradiation, apoptosis, radiosensitivity

CLC Number:

R735.1

YANG Xingxiao, ZHANG Xueyuan, ZOU Naiyi, SHAN Bao'en, MA Ming, ZHU Shuchai. Effect of HMGB1 silencing by siRNA on radiosensitivity of esophageal carcinoma KYSE30 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(4): 269-274.

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Effect of HMGB1 silencing by siRNA on radiosensitivity of esophageal carcinoma KYSE30 cells

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