晚期非小细胞肺癌中Survivin表达对顺铂敏感性和预后的预测价值

Predictive Value of Survivin on Evaluating Cisplatin Sensitivity and Prognosis of Patients with Advanced Non-small Cell Lung Cancer

目的:探讨Survivin预测晚期非小细胞肺癌(NSCLC)顺铂敏感性的价值。方法:RT-PCR、Western blot法检测人肺腺癌细胞株A549和耐药株A549/CDDP中Survivin表达,Survivin siRNA转染A549/CDDP细胞,MTT、流式细胞术检测细胞生长与凋亡;免疫组织化学和RT-PCR检测晚期NSCLC组织中Survivin mRNA和蛋白表达。结果:A549/CDDP中Survivin mRNA和蛋白表达均高于A549细胞(P<0.01);A549/CDDP细胞中Survivin siRNA+CDDP组的生长抑制率达76.4%,IC_(50)为(34.12+3.55)μg/mL,耐药指数为6.71倍,均低于对照组(P<0.05)。该组的细胞凋亡率高于对照组(P<0.01);93例NSCLC蜡块组织中,Survivin蛋白表达阴性组接受含顺铂方案化疗有效率(44.4%)高于阳性组(19.3%,P<0.01);Survivin表达阴性组总生存、无疾病进展时间较阳性组长,但差异无统计学意义(P>0.05)。Cox多因素分析结果显示Survivin蛋白表达水平不是影响预后的独立因素。20例新鲜组织中Survivin mRNA低表达组反应率(RR)(60%)和蛋白表达阴性组RR(66.7%)高于mRNA高表达组(10%)和蛋白表达阳性组(9%,P<0.05)。结论:NSCLC组织中Survivin表达阳性提示顺铂耐药,检测Survivin表达对晚期NSCLC筛选含顺铂化疗方案具有指导意义。

Objective： This study aims to investigate the expression of survivin in advanced non-small cell lung cancer （ NSCLC ）, as well as to analyze the relationship of survivin expression with cisplatin [ cis - diamminedichloroplatinum （ Ⅱ ）, CDDP ] sensitivity in vitro and in vivo. Methods： The expression of survivin mRNA and protein in A549 as well as A549/DDP cells was measured by reverse transcription-polymerase chain reaction （ RT - PCR ） and Western blot analyses, respectively. A549/CDDP cell apoptosis was detected by flow cytometry following transfection with survivin siRNA. The 3- （ 4, 5 - dimethylthiazol - 2 - yl ） -2, 5- diphenyltetrazolium bromide assay, was also performed to determine the cell viability, half-maximum inhibitory concentration （ IC50 ）, and CDDP content. The resistance index （ RI ） was then calculated. Survivin expression in advanced NSCLC tissues was detected by immunohistochemical and RT - PCR assays. Results： The expression levels of survivin mRNA and protein were both significantly higher in A549 / CDDP cells than in A549 cells （ P 〈 0.01 ）. The survivin mRNA and protein levels were markedly decreased in A549 / CDDP cells transfected with survivin siRNA. The cell growth inhibition rate in the survivin siRNA ＋ CDDP group was 76.4 %, the IC50 value was 34.12±3.55 pg/mL, and the RI was reduced to 6.7 l, which was significantly lower compared with the control group （ P 〈 0.05 ）. The apoptotic rate was 37.12 % ±3.23 % in the survivin siRNA ＋ CDDP group, which was higher compared with the control group （ P 〈 0.01 ）. The response rate （ RR ） to chemotherapy was higher in the survivin negative group than in the positive group （ 44.4 % versus 19.3 % ; P 〈 0.01 ）. Both the overall survival and time to progression were longer in the survivin-negative group than in the survivin - positive group. No significant difference was found between the two groups （ P 〉 0.05 ）. Cox multivariate regression analysis showed that the survivin expression level was not an independent prognostic factor. The RR was higher in the low - level survivin mRNA and survivin protein-negative groups than in the high-level survivin mRNA and survivin-positive groups （ 60 % versus 10 %; 66.7 % versus 9 % ）. Conclusion： Survivin overexpression contributes to the drug resistance to CDDR The high expression of survivin gene is closely associated with poor survival and lower RR. Thus, survivin is an ideal predictor of CDDP sensitivity in advanced NSCLC patients.