中文摘要：

目的构建特异性沉默CyPA基因的慢病毒颗粒，体外观察其对非小细胞肺癌H1299细胞生长的影响。方法根据已筛选的特异性沉默CyPA基因靶序列寡核苷酸，合成一对正义反义寡核苷酸DNA，采用基因克隆技术分别插入PGCL-GFP慢病毒载体，以脂质体法将重组PGCL-GFP与病毒包装载体pHelper1．0、pHelper2．0共转染293T细胞，制备假包装病毒颗粒（Lv-shCyPA）并感染非小细胞肺癌H1299细胞，4d后采用RT-PCR、Western-blot及MTT分别检测CyPA基因mRNA、蛋白表达水平及H1299细胞增殖状态。结果构建的LV-shCyPA慢病毒颗粒感染H1299细胞，CyPA mRNA及蛋白表达显著下调，H1299细胞生长明显减缓，与未转染病毒、阴性对照病毒转染的H1299细胞比较有显著差异（P〈0．05）。结论成功构建了沉默CyPA基因慢病毒载体，其产生的假包装病毒颗粒能特异地沉默CyPA基因，抑制H1299细胞增殖，提示CyPA可能在非小细胞肺癌的发生发展中发挥一定的作用。

英文摘要：

Objective To investigate the inhibitory effects of lentiviral-vector-mediated silencing of Cyclophilin A gene （CyPA） on proliferation of non-smaU cell lung carcinoma cell （NSCLC） in vitro. Methods According to the effective siRNA sequence of CyPA gene which was confirmed in our early study, the sense and antisense Oligo DNA was cloned into the pGCL-GFP vector containing green fluorescent protein （GFP） gene and expressing CyPA shRNA. The constructed lentiviral vector was identified by PCR and DNA sequencing, and was cotransfected with packing vector （ pHelper1. 0 and pHelper2. 0 ） in 293T to assemble pseudotyped virus named Lv-shCyPA. Then, non-small cell lung caracinoma lines （H1299） were transduced by Lv-shCyPA. The expression level of CyPA mRNA and protein were detected by RT-PCR and western-blot, respectively. Tumor cell proliferation was measured by MTT assay. Results The lentiviral-vector-mediated shCyPA was constructed successfully. The titer of packed virus was 1×10^7TU/ml, In the cell studies, expression of CyPA mRNA and protein were significantly decreased after transduction compared to the control groups （ P 〈 0.05 ）. Correspondingly, the proliferation of H1299 transduced by Lv-shCyPA was also obviously slowed down compared to the control groups. Conclusion The LV-shCyPA was constructed successfully, which is capable of silencing remarkably CyPA in H1299 cell and eventually resulting in the retarded tumor growth. These data suggested that CyPA gene may play a critical role in evolvement of non-small cell lung carcinoma.