摘要
目的利用已构建的带有内皮抑素基因真核表达载体质粒导入裸鼠ACHN肾细胞癌(renal cell carcinoma,RCC)细胞中,研究内皮抑素基因对RCC细胞增殖和凋亡的影响。方法ACHN细胞浓度1×107个/L,经裸鼠右侧背部皮下注射0.2 ml,共注射24只裸鼠。荷瘤裸鼠随机分为3组,每组8只。治疗组:每只裸鼠瘤内3点注射复合物100μl(内含30μl梭华-SofastTM和30μg pSecES质粒);对照组:每只裸鼠瘤内3点注射复合物100μl(内含30μl梭华-SofastTM和30μg pcDNA3.1质粒);空白组:每只裸鼠瘤内3点注射生理盐水100μl。各组每只裸鼠间隔3 d注射1次,连续注射3次。以增殖细胞核抗原(PCNA)作为细胞增殖状态,按链霉亲和素生物素法(SABC)进行免疫组化染色。细胞凋亡检测用TUNEL法。结果内皮抑素真核表达质粒能显著抑制裸鼠ACHN RCC肿瘤体积增长,pSecES治疗组平均瘤重明显小于空白组和对照组(P<0.01),与空白组的肿瘤生长抑制率为34.48%,与对照组的肿瘤生长抑制率为40.68%。治疗组肿瘤细胞的凋亡指数(AI)明显高于对照组和空白组(P<0.01)。pSecES治疗组肿瘤组织的AI/PI比值明显高于对照组和空白组(F=189.27,P<0.05)。Spearman相关分析表明,肿瘤体积与细胞增殖之间无明显相关性(r=-0.041 2,P>0.05),肿瘤体积与细胞凋亡呈负相关(r=-0.734 6,P<0.01)。结论内皮抑素基因治疗可使裸鼠ACHN RCC肿瘤细胞凋亡增加,肿瘤细胞的数量减少,在总体上表现为肿瘤的生长变慢、体积变小。
Objective To study the effect of endostatin gene therapy on proliferation and apoptosis in mouse renal cell carcinoma（RCC） cells beared in a nude mice model. Methods ACHN-bearing nude mices were treated subcutaneously with 30 μg pSecES in experimental group,30 μg pcDNA3.1 in control group and nothing in vacant group and were repeated at 3-day intervals for three times.After 34 days,the mices were sacrificed.Cell proliferation activity was detected by proliferation cell nuclear antigen（PCNA） immunostaining,and apoptosis was also detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling（TUNEL）. Results The growth of ACHN RCC cells in the experimental group were significantly suppressed than those in the control group and vacant group.The apoptotic index in the experimental group was higher than that in the control group and vacant group（both P〈0.01）.Statistical analysis revealed an inverse correlation between the apoptotic index and the tumor＇s volume（r=-0.734 6,P〈 0.01）,but the proliferation index had no correlation with the volume. Conclusions It was suggested that endostatin gene therapy may shrink the volume of the ACHN RCC tumors and slow down their growth,which indicated that this method could be employed for the treatment of renal cell carcinoma.
出处
《现代泌尿生殖肿瘤杂志》
2010年第2期108-111,共4页
Journal of Contemporary Urologic and Reproductive Oncology
关键词
内皮抑素
肾细胞癌
增殖
凋亡
基因治疗
Endostatin
Renal cell carcinoma
Proliferation
Apoptosis
Gene therapy
