p53在顺铂治疗乳腺癌引起急性肾损伤小鼠中的作用

The role of p53 in acute renal injury in mice by cisplatin used in breast cancer

目的:探讨p53/微小RNA(miRNA,miR)-205/第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因(PTEN)信号通路在顺铂诱导急性肾损伤小鼠中的作用,减轻顺铂在三阴性乳腺癌中发挥抗肿瘤活性时产生的不良反应。方法:30 mg/kg顺铂腹腔注射小鼠(购自江苏卡文斯实验动物中心)建立急性肾损伤模型。将18只雄性C57小鼠随机分为顺铂组、顺铂+p53抑制剂组、对照组。检测每组小鼠血清肌酐(Cr)和尿素氮(BUN)水平;苏木精-伊红(HE)染色评估肾小管损伤;半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3荧光染色检测肾小管上皮细胞凋亡;反转录-聚合酶链反应(RT-PCR)检测miR-205表达;蛋白质印迹法(Western blot)分析肾组织p53和PTEN表达。使用SPSS 17.0统计软件分析,采用 t检验和单因素方差分析。 结果:Western blot分析结果显示顺铂组p53和PTEN相对表达量较对照组显著升高(0.16±0.03比0.70±0.04、0.43±0.05比0.91±0.07),差异有统计学意义( t=10.792、5.625, P<0.05)。RT-PCR显示顺铂组miR-205表达量较对照组显著降低(0.96±0.02比0.41±0.03),差异有统计学意义( t=14.763, P<0.05)。与对照组比较,顺铂组显著升高Cr[(26.83±2.30) μmol/L比(92.17±3.60) μmol/L, t=15.268, P<0.05]、BUN水平[(19.67±1.86) mmol/L比(52.83±3.52) mmol/L, t=8.344, P<0.05],加重肾小管损伤分数(0.50±0.22比2.66±0.21, t=7.051, P<0.05)及肾小管上皮细胞凋亡分数(0.83±0.31比5.66±0.49, t=8.303, P<0.05),以上差异均统计学意义。顺铂+p53抑制剂组较顺铂组Cr[(92.17±3.60) μmol/L比(69.00±3.06) μmol/L, t=4.894, P<0.05]及BUN水平显著降低[(52.83±3.52) mmol/L比(40.00±2.88) mmol/L, t=2.822, P<0.05],肾小管损伤分数(2.66±0.21比1.33±0.21, t=4.471, P<0.05)及肾小管上皮细胞凋亡分数减轻(5.66±0.49比2.50±0.22, t=5.836, P<0.05),同时miR-205升高(0.41±0.03比0.61±0.03, t=9.303, P<0.05)和PTEN表达水平降低(0.91±0.07比0.65±0.04, t=3.235, P<0.05),以上差异均有统计学意义。 结论:调控p53/miR-205/PTEN轴减轻顺铂诱导急性肾损伤,为顺铂所致急性肾损伤提供潜在治疗靶点。

Objective Breast cancer susceptibility gene 1(BRCA1)mutation in triple-negative breast cancer(TNBC)get high response to cisplatin which was limited in clinical caused high ration of acute renal injury.In this report we want to explore the role of p53 in acute renal injury in mice by cisplatin used in breast cancer.Methods The model of acute renal injury was established by intraperitoneal injection of 30 mg/kg cisplatin.Eighteen male C57 mice were randomly divided into 3 groups:control group,cisplatin group,cisplatin+p53 inhibitor group.The levels of p53 and phosphatase and tensin homologue deleted on chromosome ten(PTEN)were detected by Western blotting.The levels of serum creatinine(Cr)and ureanitrogen(BUN)were measured.Renal tubular injury was estimated by hematoxylin and eosin(HE)staining.The activity of cysteinyl aspartate-specific protease(Caspase)-3 of detected by fluorescence staining.The level of microRNA(miRNA,miR)-205 was detected by reverse transcriptase-polymerase chain reaction(RT-PCR).SPSS 17.0 software was used for statistical analysis.Results In cisplatin group,the expression of p53 and PTEN was increased(0.16±0.03 vs.0.70±0.04,t=10.792;0.43±0.05 vs.0.91±0.07,t=5.625,P<0.05)and the level of miR-205 was decreased(0.96±0.02 vs.0.41±0.03,t=14.763,P<0.05)compared with control group.The same results with the level of serum of Cr[(26.83±2.30)μmol/L vs.(92.17±3.60)μmol/L,t=15.268,P<0.05],and BUN[(19.67±1.86)mmol/L vs.(52.83±3.52)mmol/L,t=8.344,P<0.05]levels,the renal tubular injury score(0.50±0.22 vs.2.66±0.21,t=7.051,P<0.05),and the apoptosis score of renal tubular epithelial cells(0.83±0.31 vs.5.66±0.49,t=8.303,P<0.05)in cisplatin group.Compared with the cisplatin group,the serum Cr[(92.17±3.60)μmol/L vs.(69.00±3.06)μmol/L,t=4.894,P<0.05]and BUN[(52.83±3.52)mmol/L vs.(40.00±2.88)mmol/L,t=2.822,P<0.05]levels,the renal tubular injury score(2.66±0.21 vs.1.33±0.21,t=4.471,P<0.05),the apoptosis score of renal tubular epithelial cells(5.66±0.49 vs.2.50±0.22,t=5.836,P<0.05)were decreased,but the level of miR-205(0.41±0.03 vs.0.61±0.03,t=9.303,P<0.05)was increased,and the level of PTEN(0.91±0.07 vs.0.65±0.04,t=3.235,P<0.05)was decreased.Conclusion Control of the axis of p53/miR-205/PTEN alleviated cisplatin induced acute renal injury in mice.