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目的 探讨反义高迁移率族蛋白1(HMGB1)基因对人胰腺癌细胞的抑制作用。方法 应用分子克隆技术构建反义HMGB1基因真核表达载体pcDNA3 1/anti HMGB1,用脂质体法将其导入人胰腺癌细胞PANC 1中,经G4 18筛选获得可稳定表达反义HMGB1的人胰腺癌细胞克隆,通过逆转录聚合酶链反应、免疫印迹法和噻唑蓝比色法检测转染4 8h后胰腺癌细胞HMGB1基因表达和体外增殖活性的变化,流式细胞仪检测细胞凋亡及细胞周期情况。结果 获得了pcDNA3 1/anti HMGB1真核表达质粒,pcDNA3 1/anti HMGB1转染可使PANC 1细胞HMGB1mRNA和蛋白表达水平显著降低(P <0 .0 1)、肿瘤细胞增殖能力明显受到抑制(P <0. 0 1) ,并出现细胞周期G1期阻滞、凋亡细胞百分数增加。结论 反义HMGB1基因的表达能有效抑制胰腺癌细胞的体外增殖并促进细胞凋亡。
Objective To investigate the inhibitory effect of anti-HMGB1 gene on human pancreatic cancer cells. Methods The antisense HMGB1 gene eukaryotic expression vector pcDNA3 1 / anti HMGB1 was constructed by molecular cloning technique and was introduced into human pancreatic cancer cell line PANC 1 by lipofectamine. The human pancreas stably expressing antisense HMGB1 was obtained by G418 screening The changes of HMGB1 gene expression and proliferation in pancreatic cancer cells were detected by reverse transcription polymerase chain reaction, Western blotting and thiazolyl blue colorimetry 48 h after transfection. Apoptosis and cell proliferation were detected by flow cytometry Periodic situation. Results The pcDNA3 1 / anti HMGB1 eukaryotic expression plasmid was obtained. The transfection of pcDNA3 1 / anti HMGB1 significantly reduced the expression of HMGB1 mRNA and protein in PANC 1 cells (P <0.01), and significantly inhibited the proliferation of tumor cells (P <0. 0 1), and cell cycle G1 arrest, the percentage of apoptotic cells increased. Conclusion The expression of antisense HMGB1 gene can effectively inhibit the proliferation and promote the apoptosis of pancreatic cancer cells in vitro.