靶向干扰骨架蛋白Transgelin抑制胰腺癌裸鼠移植瘤的生长

RNA interference targeting actin-associated protein Transgelin inhibits human pancreatic carcinoma xenograft in nude mice

目的探讨靶向干扰骨架蛋白Transgelin对人胰腺癌裸鼠移植瘤生长的影响。方法设计并体外合成靶向Transgelin的短发夹RNA（shRNA）真核表达质粒，将其转染人胰腺癌细胞BxPC3，采用RT—PCR、Western印迹法检测转染前后Transgelin表达的变化。成功造模胰腺癌裸鼠24只，按皮下注入细胞不同均分为3组：实验组（接种BxPC3／shRNA）、阴性对照组（接种空质粒转染细胞BxPC3／Neo）和空白对照组（接种未转染细胞BxPC3）。每周测量肿瘤大小，术后第28天处死所有裸鼠，计算瘤体体积，治疗前、后裸鼠体重的变化以及抑瘤率。免疫组织化学法检测移植瘤组织切片中Transgelin和增殖细胞核抗原（PCNA）蛋白的表达。结果实验组、阴性对照组和空白对照组在各时间段的肿瘤体积增长差异有统计学意义（均P〈0．05）。实验组在第21，28天肿瘤体积显著小于阴性对照组和空白对照组（均P〈0．05）。实验组第28天瘤重为（0．74±0．21）g，阴性对照组为（1．42±0．28）g，空白组为（1．59±0．24）g。实验组瘤重与阴性、空白对照组相比差异均有统计学意义（均P〈0．05），抑瘤率达53．5％。实验组PCNA指数显著低于阴性对照组和空白对照组（均P〈0．05）。结论靶向封闭Transgelin基因能明显抑制胰腺癌细胞BxPC3在裸鼠体内的增殖及肿瘤生长。

Objective To investigate the in vivo effect of silenced actin-associated protein Transgelin on the growth of human pancreatic carcinoma xenograft in nude mice. Methods Human pancreatic cancer cell line BxPC3 were transfected with small hairpin RNA （shRNA） eukaryotic expression vector targeting Transgelin gene. RT-PCR and Western blot were used to analyze Transgelin expression after transfection. 24 animal models were randomly divided into three groups with 8 in each： Experimental group （transplanted BxPC3/Transgelin shRNA） , negative control group （transplanted BxPC3/Neo） and untreated group （transplanted BxPC3 ）. Tumor size was measured weekly. All mice were sacrificed after 28 days. Tumor volume was calculated, inhibitory effect was analyzed. Immunohistochemical staining of paraffin sections for Transgelin and proliferating cell nuclear antigen （PCNA） proteins were performed. Results Tumors varied in sizes among 3 groups （ all P 〈 0.05 ）. On day 21 and 28 tumor was significantly smaller in experimental group than those in control groups. Tumor weighed（0. 74 ±0. 21 ） g in experimental group, lower than that in negative control group（ 1.42 ±0. 28） g and untreated group（ 1.59 ±0. 24） g （ all P 〈 0. 05 ）. The inhibitory effect was 53.5% in experimental group. The PCNA index was significantly lower in experimental group than those in control groups （ all P 〈 0. 05 ）. Coneluslons Deletion of Transgelin gene can significantly inhibit the proliferation and tumor growth of BxPC3 ceils in nude mice.