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目的:Livin是近年来发现的人类凋亡抑制蛋白(inhibitorof apoptosis protein,IAP)家族的新成员,发现在多数肿瘤中表达,与肿瘤发生有密切关系,并可能作为肿瘤诱导凋亡治疗的新靶点。本研究旨在探讨si RNA-Livin和夫拉平度(Flavopiridol,FP)协同凋亡诱导配体(TRAIL)两种方式有效抑制Livin表达,诱导非小细胞肺癌SPC-A1凋亡并增强对化疗药物顺铂的敏感性。方法:si RNA-Livin转染SPC-A1,Real-Time PCR检测Livin基因的表达水平,MTT检测干扰组和干扰加药组肿瘤细胞的增殖及活性;TRAIL、FP单独及联合作用诱导细胞凋亡,蛋白质印迹法检测凋亡抑制蛋白Livin的表达水平,MTT检测各处理组细胞的增殖及活性。结果:100 nmol/LFP处理组(F)细胞存活率为(84.30±1.34)%,100 ng/m LTRAIL处理组(T)为(93.40±1.56)%,FP和TRAIL联合组(F+T)为(48.02±1.35)%,si RNA-Livin处理组为(50.88±1.14)%,1.2μg/m L Cisplatin处理组为(19.30±0.89)%,si RNA-livin+Cisplatin组为(14.37±0.81)%,FP+T+Cisplatin组为(10.86±0.87)%,C组存活率为100%。F+T组对细胞的增殖抑制作用显著高于单独用药组,si RNA-livin+Cisplatin与si RNA-Livin组相比、FP+T+Cisplatin与FP+T组相比都显著增强了化疗药物对SPC-A1的杀伤作用。50μmol/LZ-VAD-FMK预处理后联合用药组细胞的存活率为(88.16±1.64)%,caspase抑制剂能明显抑制F+T联合处理组的凋亡效应。结论:RNA干扰和F+T联合用药都能显著降低凋亡抑制蛋白Livin的表达,有效抑制肿瘤细胞的增殖生长,并增强肿瘤细胞对化疗药物顺铂的敏感性,为肺腺癌的靶向治疗提供新的理论依据。
OBJECTIVE: Livin is a new member of the inhibitor of apoptosis protein (IAP) family found in recent years. It has been found that Livin is closely related to tumorigenesis in most tumors and may serve as a new target for tumor-induced apoptosis point. The purpose of this study is to investigate whether si RNA-Livin and Flavopiridol (FP) cooperate with apoptosis-inducing ligand (TRAIL) in two ways to effectively inhibit Livin expression, induce apoptosis of non-small cell lung cancer SPC-A1 and enhance chemosensitivity Platinum sensitivity. Methods: The expression of Livin gene was detected by real-time PCR with si RNA-Livin. The proliferation and activity of tumor cells were detected by MTT assay. The apoptosis of the cells was induced by TRAIL and FP alone and in combination The expression of Livin was detected by Western blotting. The proliferation and activity of cells in each treatment group were detected by MTT assay. Results: The survival rate of the cells treated with 100 nmol / LFP was (84.30 ± 1.34)% and that of the cells treated with 100 ng / m LTRAIL was (93.40 ± 1.56)%, while the combination of FP and TRAIL (F + T) (48.02 ± 1.35)%, (50.88 ± 1.14)% in si RNA-Livin group and (19.30 ± 0.89)% in 1.2 μg / ml Cisplatin group and (14.37 ± 0.81) )%, FP + T + Cisplatin group (10.86 ± 0.87)%, C group survival rate was 100%. The inhibitory effect of F + T on the cell proliferation was significantly higher than that of the untreated group, and compared with the si RNA-Livin group, both the si RNA-livin + Cisplatin and the FP-T group significantly enhanced the chemotherapeutic drugs SPC-A1 killing effect. The survival rate of the combination group treated with 50μmol / L Z-VAD-FMK was (88.16 ± 1.64)%, and caspase inhibitor significantly inhibited the apoptosis of F + T combined treatment group. CONCLUSION: Both RNA interference and F + T combined treatment can significantly reduce the expression of Livin, effectively inhibit the proliferation and proliferation of tumor cells and enhance the sensitivity of tumor cells to chemotherapeutic drug cisplatin, which is the target of lung adenocarcinoma Treatment provides a new theoretical basis.