期刊文献+

RNA干扰沉默瘦素基因对人肺腺癌A549细胞裸鼠移植瘤生长的影响

Effects of Leptin gene down-regulation by RNA interference on xenograft of human lung adenocarcinoma A549 cell line in nude mice
原文传递
导出
摘要 目的利用RNA干扰技术沉默人肺腺癌A549细胞中的瘦素(Leptin)基因的表达,探讨Leptin基因表达沉默对人肺腺癌A549细胞裸鼠皮下移植瘤生长的影响。方法以慢病毒为载体,将携带siRNA的Leptin基因及空质粒转染人肺腺癌A549细胞,转染成功后将转染Leptin-siRNA的人肺腺癌A549细胞(干扰组)、转染空质粒的人肺腺癌A549细胞(阴性对照组)及未转染任何序列的人肺腺癌A549细胞(空白对照组)分别接种于BALB/C裸鼠右腋窝皮下,构建裸鼠皮下移植瘤模型,观察各组细胞在裸鼠体内的成瘤率、瘤体体积、瘤体重量,绘制肿瘤生长曲线并计算抑瘤率。免疫组织化学及免疫蛋白印迹分别检测瘤体组织中Leptin蛋白表达情况,原位末端凋亡法检测移植瘤组织凋亡情况。结果干扰组与空白对照组及阴性对照组相比,裸鼠移植瘤瘤体体积、瘤体重量明显降低,其体积抑瘤率为(59.3±3.1)%,瘤重抑瘤率为(64.5±4.8)%,差异均有统计学意义(F=9.118、9.121,P值均〈0.05)。干扰组Leptin蛋白表达较空白对照组及阴性对照组明显减少,差异有统计学意义(F=13.784,P〈0.05)。干扰组沉默Leptin基因后细胞凋亡增多,凋亡率为(47.3±4.1)%,明显高于空白对照组[(5.6±1.5)%]及阴性对照组[(7.2±2.8)%],差异有统计学意义(F=14.537,P〈0.05)。结论沉默人肺腺癌A549细胞中的Leptin基因表达可以有效抑制裸鼠皮下移植瘤的生长,Leptin基因有望成为肺腺癌基因治疗的一个新靶点。 Objective To investigate the inhibitory effect of Leptin gene in vivo silencing by RNA interference on xenograft of human lung adenocarcinoma A549 cell line in nude mice. Methods The Leptin gene and plasmid carrying siRNA were transfected into A549 cells by lentivirus. Three different 13ALB/C nude mice models were established through subcutaneous injection of A549 cells into three nude mice groups : the blank group was inculated with blank A549 cells, while the negative group was inoculated with cells transfected with empty plasmid, the RNA inteference group was inoculated with cells transfected with Leptin-siRNA. The growth of tumors was observed, and the volume and weight of the tumors were measured at different time points. The curve of tumor growth was described, and the inhibition rate was calculated. Leptin gene expression in tumor tissues was determined by inmunohistochemistry and Western blot. Cell apoptosis of tumor was detected by terminal-deoxynuele oitidyl tramsferase mediated nick end labeling. Results Smaller volume and lighter tumor weight were observed in the interference group compared to the blank and negative groups ( F = 9. 118, 9. 121, both P〈0. 05). The inhibition rate of tumor volume was (59.3 ±3.1) %, and the inhibition rate of tumor weight was (64.5 ±4.8) %. Leptin protein expression level was also significantly lower than the blank and negative groups ( F =13.784, P 〈0.05). The apoptosis rate in the interference group was higher than in the blank and negtive groups [(47.3±4.1) %,(5.6±1.5),(7.2±2.8)%, F =14.537, P 〈0.05].Conclusions Silencing the expression of Leptin gene can inhibit the growth of subcutaneous xenograft tumor effectively. It is expected that Leptin gene would be a new target for gene therapy in lung adenocarcinoma.
作者 承柯伟 杨明夏 俞小卫 Cheng Kewei;Yang Mingxia;Yu Xiaowei.(Department of Respiratory and Critical Care Medicine, Changzhou NO. 2 People's Hospital, the Affiliated Hospital of Nanjing Medical University, Changzhou 213000, China)
出处 《国际呼吸杂志》 2018年第12期881-885,共5页 International Journal of Respiration
关键词 瘦素 RNA干扰 A549细胞 移植瘤 Leptin RNA interference A549 cells Transplantation tumor
  • 引文网络
  • 相关文献

参考文献3

二级参考文献39

  • 1胡斐,效小莉,濮德敏,邹倩,李天.瘦素及其受体在宫颈癌中的表达和意义[J].中国组织化学与细胞化学杂志,2006,15(3):297-301. 被引量:5
  • 2薛秀兰,林菊生,宋宇虎,蔡晓坤,周鹤俊.靶向leptin基因的siRNA对大鼠肝星状细胞生物学特性的影响[J].华中科技大学学报(医学版),2006,35(3):331-335. 被引量:5
  • 3Ribeiro R, Araujo A, Lopes C, et al. Immunoinflammatory mechanisms in lung cancer development is leptin a mediator? J Thorac Oneol, 2007,2 : 105-108.
  • 4Huang CY, Yu HS, Lai TY,et al. Leptin increases motility and integrin up-regulation in human prostate cancer cells. J Cell Physiol,2011,226 : 1274-1282.
  • 5Uddin S, Bu R, Ahmed M, et al. Overexpression of leptin receptor predicts an unfavorable outcome in Middte Eastern ovarian cancer. Mol Cancer, 2009,8 : 74.
  • 6Kim SH, Nagalingam A, Saxena NK, et al Benzyl isothiocyanate inhibits oncogenic actions of leptin in human breast cancer cells by suppressing activation of signal transducer and activator of transcription 3. Carcinogenesis, 2011,32 : 359-367.
  • 7Ogunwobi OO, Beales IL. Globular adiponectin, acting via adiponectin receptor-1, inhibits leptin-stimulated oesopbageal adenocarcinoma cell proliferation. Mol Cell Endocrinol, 2008, 285,43-50.
  • 8Chen H, Xia M, Lin M, et al. Role of methionine adenosyltransferase 2A and S-adenosylmethionine in mitogen induced growth of human colon cancer cells. Gastroenterology, 2007,133 : 207-218.
  • 9Jemal A, Siegel R, Xu J, et al. Cancer statistics, 2010. CA Cancer J Clin, 2010,60 : 277-300.
  • 10Hanagiri T, Baba T, So T, et al. Time trends of surgical outcome in patients with non small eell lung caneer. J Thorac Oncol, 2010,5 : 825-829.

共引文献12

;
使用帮助 返回顶部