摘要
目的探讨沉默Annexin A2基因对放射抗拒鼻咽癌细胞裸鼠移植瘤放射敏感性的影响。方法选择36只SPF级BALB/c裸鼠,雄性,6周龄,体质量18~22 g。按随机数字表法分为对照组[CNE-2(R743)组]、单纯照射组[CNE-2(R743)/R组]、转染对照组(sh R-C组)、转染对照联合照射组(sh R-C/R组)、转染组(sh R-ANXA2组)及转染联合照射组(sh R-ANXA2/R组),每组6只。用p Gene Clip空载体及p Gene Clip-ANXA2-sh RNA载体转染CNE-2(R743)细胞构建移植瘤动物模型,用6 MV X射线在室温下行单次照射动物,总吸收剂量为10 Gy,剂量率为每分钟400 c Gy,照射野面积为20 cm×20 cm。观察各组裸鼠移植瘤照射后的生长情况、移植瘤体积变化情况,免疫组织化学检测移植瘤组织中Annexin A2的表达,Western blot检测移植瘤组织中Ku70和增殖细胞核抗原(PCNA)的表达。结果 10 Gy X射线照射后3周,sh R-ANXA2/R组(168.46%±129.00%)裸鼠肿瘤生长速度最慢,与CNE-2(R743)/R组(462.26%±195.60%)和sh R-C/R组(407.99%±116.60%)比较,差异有统计学意义(P<0.05)。沉默Annexin A2的sh R-ANXA2/R组(2.68%±1.29%)的相对生长速率较CNE-2(R743)/R组(5.62%±1.96%)和sh R-C/R组(5.08%±1.17%)降低(P<0.05),生长抑制率增加。sh R-ANXA2/R组的q值为1.06,提示有放射增敏作用。免疫组织化学结果显示,sh R-ANXA2组移植瘤中Annexin A2的表达较CNE-2(R743)组和sh R-C组低。各组裸鼠移植瘤组织中Ku70蛋白的表达未见明显变化(P>0.05)。sh R-ANXA2/R组(2.97±0.44)的PCNA蛋白表达水平较CNE-2(R743)/R组(2.04±0.63)和sh R-C/R组(1.26±0.38)明显升高(P<0.05)。结论沉默Annxin A2基因可以提高裸鼠体内鼻咽癌移植瘤的放射敏感性。
Objective To investigate the effect of silencing Annexin A2 gene on radiosensitivity in nude mice xenografted with radioresistant nasopharyngeal carcinoma cells. Methods Thirty-six SPF grade BALB/c nude mice（males aged 6-week-old with body weight of 18-22 g） were used. All mice were randomly divided into 6 groups（n = 6） by random number table： control group[CNE-2（R743） group], irradiation group[CNE-2（R743）/R group], transfection control group（sh R-C group）, transfection control combined irradiation group（sh R-C/R group）, transfection group（sh R-ANXA2 group） and transfection combined irradia-tion group（sh R-ANXA2/R group）. The p Gene Clip empty vector and p Gene Clip-ANXA2-sh RNA vector transfected CNE-2（R743）cells were used to produce transplanted animal models, and then irradiated with 6 MV X-ray at room temperature. The total dose of 10 Gy at dose rate of 400 c Gy per minute was administered, and the irradiation field area was 20 cm × 20 cm. The growth of xenograft and changes of tumor volume in nude mice for each group were observed after irradiation, the immunohis-tochemistry was used to detect expression of Annexin A2 in xenograft tissues, and expression of Ku70 and proliferating cell nuclear antigen（PCNA） in xenograft tumor tissues were detected by Western blot. Results After 10 Gy X-ray irradiation, the tumor growth rate of sh R-ANXA2/R group[（168.46 ± 129.00） %] was the slowest compared with that of CNE-2（R743）/R group[（462.26 ± 195.60） %] and sh R-C/R group[（407.99 ± 116.60） %], and the difference was statistically significant（P〈0.05）. The relative growth rate of sh R-ANXA2/R group [（2.68 ± 1.29） %] was lower than that of CNE-2（R743）/R group[（5.62 ± 1.96） %]and sh R-C/R group[（5.08 ± 1.17） %]（P〈0.05）, and growth inhibition rate increased. The q value of sh R-ANXA2/R group was 1.06,which suggested radiosensitizing effect. The immunohistochemical results showed that the expression of Annexin A2 in xenograft of sh R-ANXA2/R group was lower than that of CNE-2（R743） and sh R-C group. The expression of Ku70 protein in xenograft of mice was no changed significantly（P〈0.05）. The expression level of PCNA protein in sh R-ANXA2/R group[（2.97 ± 0.44）] was significantly higher than that in CNE-2（R743）/R group[（2.04 ± 0.63）] and sh R-C/R group[（1.26 ± 0.38）]（P〈0.05）. Conclusion It is demonstrated that silencing Annxin A2 gene could improve the radiosensitivity of nasopharyngeal carcinoma xenografts in nude mice.
出处
《生物医学工程与临床》
CAS
2017年第6期646-652,共7页
Biomedical Engineering and Clinical Medicine
基金
福建省科技计划重点项目(2014Y0014)
国家临床重点专科建设项目[国卫办医函(2013)554号]
福建省临床重点专科建设项目资助(西医类别)[闽卫科教(2012)149号]
