Department of Radiotherapy, Qilu Hospital of Shandong University, No. 107, Wenhua West Road, Lixia District, Jinan, Shandong Province, P.R. China.
Department of Radiology, Qilu Hospital of Shandong University, No. 107, Wenhua West Road, Lixia District, Jinan, Shandong Province, P.R. China.
J Radiat Res. 2019 May 1;60(3):318-327. doi: 10.1093/jrr/rrz003.
The aim of this study was to evaluate the effect of SERPINB2 on cell proliferation, cell cycle, epithelial-mesenchymal transition (EMT), invasion, migration, and radiosensitivity in nasopharyngeal carcinoma cells. Both CNE2R and CNE2 cells were transfected with pEGFP-N1-SERPINB2. Cell proliferation was measured by MTT assay, cell cycle by flow cytometry, and SERpINB2 expression by quantitative real-time polymerase chain reaction (qRT-PCR). Western blot was carried out to detect the protein expression. In addition, SERPINB2 and β-catenin were located intracellularly using an immunofluorescent assay, and cell migration and invasion were measured by wound healing and Transwell assays, respectively. Radiosensitivity was assessed using colony formation and MTT assays. SERPINB2 expression was downregulated in CNE2R cells. After transfection with pEGFP-N1-SERPINB2, the OD values were decreased, and there was an increased fraction in the G2/M phase. Moreover, SERPINB2 overexpression could inhibit the invasion and migration capabilities of CNE2R and CNE2 cells, with downregulation of vimentin, N-cadherin, nuclear β-catenin, matrix metalloproteinase (MMP)-2 and MMP-9, and upregulation of E-cadherin. Moreover, transfection with the SERPINB2 plasmid reduced the growth rate of CNE2R cells at doses of 2, 4 and 6 Gy, and also decreased the surviving fractions. Overexpression of SERPINB2 could reduce the proliferation, invasion and migration capabilities of CNE2R and CNE2 cells, and led to G2/M arrest via EMT inhibition, and this may be a potential strategy for enhancing the radiation sensitivity of nasopharyngeal carcinoma cells.
本研究旨在评估丝氨酸蛋白酶抑制剂 B2(SERPINB2)对鼻咽癌细胞增殖、细胞周期、上皮-间质转化(EMT)、侵袭、迁移和放射敏感性的影响。分别用 pEGFP-N1-SERPINB2 转染 CNE2R 和 CNE2 细胞。通过 MTT 法检测细胞增殖,流式细胞术检测细胞周期,实时定量聚合酶链反应(qRT-PCR)检测 SERpINB2 表达。Western blot 检测蛋白表达。此外,通过免疫荧光法检测 SERPINB2 和 β-连环蛋白的细胞内定位,通过划痕愈合和 Transwell 测定分别检测细胞迁移和侵袭。通过集落形成和 MTT 测定评估放射敏感性。CNE2R 细胞中 SERPINB2 表达下调。转染 pEGFP-N1-SERPINB2 后,OD 值降低,G2/M 期细胞比例增加。此外,SERPINB2 过表达可抑制 CNE2R 和 CNE2 细胞的侵袭和迁移能力,下调波形蛋白、N-钙黏蛋白、核 β-连环蛋白、基质金属蛋白酶(MMP)-2 和 MMP-9,上调 E-钙黏蛋白。此外,转染 SERPINB2 质粒可降低 CNE2R 细胞在 2、4 和 6 Gy 剂量下的生长速度,并降低存活分数。SERPINB2 的过表达可降低 CNE2R 和 CNE2 细胞的增殖、侵袭和迁移能力,并通过 EMT 抑制导致 G2/M 期阻滞,这可能是增强鼻咽癌细胞放射敏感性的一种潜在策略。