rAAV2-slug-siRNA对原位胰腺癌移植瘤转移和血管生成影响的实验研究

Influence of rAAV2-slug-siRNA-situ on metastasis and angiogenesis pancreatic tumor in rats

目的:研究slug基因的RNA干扰重组腺病毒载体(rAAV2-slug-siRNA)对原位胰腺癌移植瘤的转移和血管生成的影响。方法:建立胰腺癌细胞株AsPC-1裸鼠原位胰腺癌移植瘤模型,随机分成3组,每组8只,分为空白对照组(腹腔注射生理盐水)、阴性对照组(腹腔注射rAAV2-GFP)、实验组(腹腔注射rAAV2-slug-siRNA)。10周后利用CO2麻醉法处死裸鼠,观察原位胰腺癌移植瘤的重量,抑瘤率,肝、胃肠、腹腔转移,腹水等情况,及肿瘤细胞微血管密度(MVD)。RT-PCR检测移植瘤的slug mR-NA表达,Western blot检测slug蛋白的表达。结果:胰腺癌细胞株AsPC-1裸鼠原位胰腺癌移植瘤模型建立成功,成瘤率100%。实验组原位胰腺癌移植瘤瘤重明显低于阴性对照组(P<0.05),抑瘤率为70.83%。阴性对照组诱发的裸鼠胰腺癌质地硬,肿块向四周浸润并形成癌性粘连,与阴性对照组相比,实验组腹膜、肝、毗邻脏器转移率和形成腹水率均明显下降(P<0.05)。实验组MVD明显少于阴性对照组(P<0.05),slug mRNA相对表达量(RT-PCR)和slug蛋白相对表达量(Western blot)明显低于阴性对照组(P<0.05)。结论:rAAV2-slug-siRNA可能通过抑制肿瘤血管生成而抑制原位胰腺癌移植瘤转移。

Objective： To investigate the influence of RNA interference slug gene recombinant adenovirus vector（rAAV2-slug-siRNA） on metastasis and angiogenesis of orthotopic pancreatic tumor.Methods：Orthotopic rat models of pancreatic cancer induced by pancreatic cancer cell line AsPC-1 were established and randomly divided into 3 groups with 8 rats in each group.Control group was treated with normal saline being injected to abdominal cavity,the negative control group with rAAV2-GFP,while the experiment group with rAAV2-slug-siRNA.Rats were sacrificed 10 weeks after the treatment,and situ pancreatic tumor weight,inhibition rate,metastasis in liver,gastrointestine,peritoneal metastasis,ascites,as well as microvessel density of tumor cells（MVD） in each group were observed.Expression of slug mRNA,and slug protein were assessed by RT-PCR and Western blot detection,respectively.Results： Orthotopic rat models of pancreatic cancer induced by pancreatic cancer cell line AsPC-1 were established successfully with a success rate of 100%.Percentage of rats with liver metastasis,peritoneal metastasis,metastasis in adjacent organs or ascites in the experiment group was significant lower than that in the negative control group（P 0.05）,showing tumor inhibition rate of 70.83%.The induced tumor of the negative control group showed characters of hard texture,infiltration and adhesions.MVD of the experimental group was significant less than that of the negative control group（P0.05）.The expression level of both slug mRNA and slug protein of the experiment group was lower than that of the negative control group（P0.05）.Conclusions：rAAV2-slug-siRNA may inhibit orthotopic pancreatic tumor metastasis by inhibiting angiogenesis.