目的 探讨帕比司他通过靶向ADP核糖基化因子样蛋白4C(ARL4C)对肾癌细胞增殖、迁移和侵袭的影响.方法 将肾癌细胞株786-O分为空白组(用磷酸盐缓冲溶液干预)、实验组(用50 nmol·L帕比司他干预)、空载体组(先转染空载体,然后用磷酸盐缓冲溶液干预)、过表达组(先转染ARL4C过表达载体,然后用磷酸盐缓冲溶液干预)、联合组(先转染ARL4C过表达载体,然后用50 nmol·L-1帕比司他干预).用噻唑蓝法和划痕实验检测细胞的增殖、迁移和侵袭水平,用酶标仪检测细胞内胆固醇水平,用实时荧光定量聚合酶链反应和蛋白质印迹法检测ARL4C mRNA和蛋白的表达水平.结果 空白组、实验组、空载体组、过表达组和联合组的细胞增殖率分别为(100.00±0)％、(61.08±5.82)％、(101.22±5.92)％、(121.94±6.63)％和(101.78±6.87)％,迁移率分别为(44.59±2.49)％、(29.02±2.09)％、(42.75±2.42)％、(54.19±3.05)％和(41.91±2.75)％,侵袭能力分别为(264.50±6.52)、(182.70±5.83)、(257.80±7.91)、(322.40±8.27)和(266.80±8.15)个,胆固醇水平分别为(72.48±6.21)、(36.48±3.44)、(73.89±5.91)、(89.21±8.89)和(73.06±6.92)mmol·L-1,ARL4C mRNA 相对表达水平分别为 1.23±0.22、0.24±0.08、1.27±0.25、1.67±0.38 和 1.27±0.25,ARL4C 蛋白相对表达水平分别为 1.06±0.03、0.17±0.04、1.03±0.05、1.37±0.18 和 1.05±0.08.实验组的上述指标和空白组比较,过表达组的上述指标与空白组和空载体组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 帕比司他通过下调ARL4C表达,降低细胞内胆固醇水平,从而抑制肾癌细胞的增殖、迁移和侵袭水平.
Objective To explore the effects of panobinostat on the proliferation,migration and invasion of renal carcinoma cells via targeting ADP ribosylated factor-like protein 4C(ARL4C).Methods According to different treatments,human renal carinoma 786-O cells were divided into blank group(phosphate buffer treatment),experimental group(50 nmol·L-1 panobinostat treatment),empty vector group(transfection with empty vector and no treatment),overexpression group(transfection with ARL4C overexpression vector and no treatment)and combined group(transfection with ARL4C overexpression vector and 50 nmol·L-1 panobinostat treatment).The proliferation,migration and invasion of cells were detected by methyl thiazolyl tetrazolium assay and scratch assay.Cholesterol transport levels in cells were detected by enzyme-labeled assay.The mRNA and protein levels of ARL4C in cells were detected by real-time fluorescence quantitative polymerase chain reaction and Western blot.Results The indicators of cell proliferation in blank group,experimental group,empty vector group,overexpression group and combined group were(100.00±0)％,(61.08±5.82)％,(101.22±5.92)％,(121.94±6.63)％and(101.78±6.87)％;the indicators of cell migration were(44.59±2.49)％,(29.02±2.09)％,(42.75±2.42)％,(54.19±3.05)％and(41.91±2.75)％;the indicators of cell invasion were 264.50±6.52,182.70±5.83,257.80±7.91,322.40±8.27 and 266.80±8.15;the intracellular levels of cholesterol were(72.48±6.21),(36.48±3.44),(73.89±5.91),(89.21±8.89)and(73.06±6.92)mmol·L-1;the relative expression levels of ARL4C mRNA were 1.23±0.22,0.24±0.08,1.27±0.25,1.67±0.38 and 1.27±0.25;the relative expression levels of ARL4C protein were 1.06±0.03,0.17±0.04,1.03±0.05,1.37±0.18 and 1.05±0.08,respectively.Between the blank group and experimental group,the above indicators have statistically significant differences(all P＜0.05);compared with the blank group and empty vector group,the above indicators in overexpression group have statistically significant differences(all P＜0.05).Conclusion Panobinostat downregulate the intracellular levels of cholesterol by reducing the expression level of ARL4C in 786-O cells,thus inhibiting the proliferation,migration and invasion of renal carcinoma cells.
杨志云;王雪莉;田秀岭
商丘医学高等专科学校临床医学院,河南商丘 476000商丘市中心医院肿瘤科,河南商丘 476002
药学
帕比司他;ADP核糖基化因子样蛋白4C;肾癌;细胞增殖;胆固醇
panobinostat;ADP ribosylation like factor 4C;renal cancer;cell proliferation;cholesterol
《中国临床药理学杂志》 2024 (011)
1569-1572 / 4
河南省科技攻关指导基金资助项目(232102310516)
10.13699/j.cnki.1001-6821.2024.11.006
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