目的：运用RNA干扰（RNA interference，RNAi）技术沉默结肠癌细胞株LoVo中Livin基因的表达，研究Livin对LoVo细胞增殖和凋亡等生物学特性的影响。 方法： 构建针对Livin基因的干扰质粒pSilencer4.1-L1和pSilencer4.1-L2，转染LoVo细胞，通过RT-PCR、Western blotting检测转染后LoVo细胞Livin的表达，并用流式细胞术、克隆形成实验、MTT法检测转染后LoVo细胞的凋亡、细胞周期、克隆形成、增殖以及对顺铂的敏感性。 结果： 成功构建pSilencer4.1-L1、pSilencer4.1-L2干扰质粒，pSilencer4.1-L1可有效沉默LoVo细胞中Livin mRNA和蛋白的表达（P<0.01），但pSilencer4.1-L2效果不明显。与对照组相比，pSilencer4.1-L1转染后LoVo细胞的凋亡率增加\[（24.2±3.2）% vs（8.1±1.4）%，P<0.01\]；细胞增殖明显抑制，转染后72 h增殖抑制率约70%；细胞的克隆形成率明显下降\[（15±4.6）% vs（85±5.8）%，P<0.01\]；S期细胞明显增加\[（45.7±49）% vs（28.0±3.0）%，P<0.01\]，G1期细胞明显减少\[（43.0±5.2）% vs（62.8±5.1）%，P<0.01\]；对顺铂的敏感性增加，细胞凋亡率增加\[（65.3±6.2）% vs（43.4±6.9）％，P<0.01\]。 结论： pSilencer4.1-L1可有效沉默结肠癌LoVo细胞中Livin基因的表达，抑制细胞增殖和克隆形成、诱导细胞凋亡、增加细胞对顺铂的敏感性。

Objective :To observe the effects of Livin on biology characteristics, such as proliferation and apoptosis, of colon cancer LoVo cells by RNA interference (RNAi) targeting Livin gene. Methods: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 targeting Livin gene were constructed and transfected into LoVo cells. Then the expression of Livin was detected by RT-PCR and Western blotting. And the apoptosis, cell cycle, colony formation, proliferation of LoVo cells, as well as sensitivity of LoVo cells to cisplatin, were detected by flow cytometry, colony formation assay, and MTT. Results: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 were successfully constructed. pSilencer4.1-L1, but not pSilencer4.1-L2, effectively silenced the Livin mRNA and protein expression in LoVo cells (P<0.01). Compared with the control group, LoVo cells in pSilencer4.1-L1 transfection group showed an increased apoptosis rate (\[24.2±32\]% vs \[8.1±1.4\]%, P＜0.01), a decreased cell proliferation (inhibitory rate: about 70% after 72 h), a decreased colony formation rate (\[15±4.6\]% vs \[85±5.8\]%, P＜0.01), increased S phase cells (\[45.7±49\]% vs \[28.0±3.0\]%, P＜0.01), decreased G1 phase cells (\[43.0±5.2\]% vs \[62.8±5.1\]%, P＜0.01), and an increased sensitivity to cisplatin (apoptosis rate increased from \[43.4±6.9\]% to \[65.3±6.2\]%, P＜0.01). Conclusion: pSilencer4.1-L1 can effectively silence Livin gene expression in colon cancer LoVo cells, inhibit proliferation and colony formation, induce apoptosis, and enhance sensitivity of LoVo cells to cisplatin.

西安市科技攻关计划项目\[No. HM1124(2)\]