›› 2010, Vol. 30 ›› Issue (7): 763-.

• 论著(基础研究) • 上一篇    下一篇

CpG寡聚脱氧核苷酸抑制肾癌细胞增殖及肿瘤生长的体内外实验研究

马斌斌, 周佩军, 徐 达, 王祥慧, 邵 琨   

  1. 上海交通大学 医学院瑞金医院泌尿外科, 上海 200025
  • 出版日期:2010-07-25 发布日期:2010-07-26
  • 通讯作者: 周佩军, 电子信箱: peijunzhou@yahoo.com;徐 达, 电子信箱: daxue@medmail.com。
  • 作者简介:马斌斌(1983—), 男, 硕士生;电子信箱: jerry_ma1983@hotmail.com。

In vivo and in vitro study of inhibition effects of CpG oligodeoxynucleotide on renal carcinoma cell proliferation and tumor growth

MA Bin-bin, ZHOU Pei-jun, XU Da, WANG Xiang-hui, SHAO Kun   

  1. Department of Urology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2010-07-25 Published:2010-07-26

摘要:

目的 研究CpG寡聚脱氧核苷酸(CpG ODN)在体内外对肾癌细胞增殖及肿瘤生长的抑制作用,并探讨其可能的作用机制。方法 采用CCK-8法检测不同浓度CpG ODN1826和non-CpG ODN1982 (ODN1982)干预对体外培养人肾透明细胞癌Caki-1细胞增殖的影响。24只BALB/c裸鼠皮下接种Caki-1细胞建立荷瘤裸鼠模型,4 d后根据每周1次的治疗方式随机分为对照组(PBS)、ODN1982组(50 μg ODN1982)、小剂量CpG ODN1826组(25 μg CpG ODN1826)和大剂量CpG ODN1826组(50 μg CpG ODN1826),每组6只。于建模后35 d 处死动物,比较各组荷瘤裸鼠的肿瘤体积、质量及肿瘤组织学改变;体外分离各组荷瘤裸鼠脾淋巴细胞,乳酸脱氢酶释放法检测不同靶标比例(脾淋巴细胞∶Caki-1细胞或脾淋巴细胞∶小鼠淋巴瘤细胞YAC-1细胞)的荷瘤裸鼠脾淋巴细胞对肿瘤细胞的杀伤活性。结果 ODN1982对Caki-1细胞的体外增殖活性无显著影响;500 μg/mL CpG ODN1826干预可显著抑制Caki-1细胞增殖。至实验终止时点,小剂量和大剂量CpG ODN1826组荷瘤裸鼠的肿瘤体积显著小于ODN1982组和对照组 (P<0.05);各组间肿瘤质量比较差异均具有统计学意义(P<0.05);组织学观察显示,小剂量和大剂量CpG ODN1826组肿瘤组织较多炎性细胞浸润。当靶标比例为50:1时,大剂量CpG ODN1826组裸鼠脾淋巴细胞对Caki-1细胞和YAC-1细胞的杀伤活性分别为(9.74±1.16)%和(79.65±5.23)%,显著高于对照组的(7.67±0.22)%和(10.12±3.03)%及ODN1982组的(7.66±0.93)%和(27.60±9.83)%(P<0.05)。结论 CpG ODN对体外肾癌细胞增殖及荷瘤裸鼠肿瘤组织生长均具有明显抑制作用,其机制可能与CpG ODN增强荷瘤裸鼠的固有免疫应答有关。

关键词: CpG寡聚脱氧核苷酸, 肾癌, 固有免疫

Abstract:

Objective To investigate the inhibition effects of CpG oligodeoxynucleotide (CpG ODN) on renal carcinoma cell proliferation and tumor growth in vivo and in vitro, and explore the possible mechanism. Methods The effects of different concentrations of CpG ODN1826 and non-CpG ODN1982 (ODN1982) interventions on proliferation of human renal clear cell carcinoma cell line Caki-1 cultured in vitro were determined with CCK-8 method. Tumor-bearing nude mouse models were established by subcutaneous implantation of Caki-1 cells into 24 BALB/c mice, and were randomly divided into control group (PBS), ODN1982 group (50 μg ODN1982), low-dose CpG ODN1826 group (25 μg CpG ODN1826) and high-dose CpG ODN1826 group (50 μg CpG ODN1826) 4 d later according to means of weekly treatment, with 6 mice in each group. Mice were sacrificed 35 d after model establishment, and changes of tumor volumes, tumor weight and tumor histology were compared among groups. Splenic lymphocytes of tumor-bearing mice in each group were isolated in vitro, and lactate dehydrogenase release method was employed to detect the cytolytic activity of splenic lymphocytes of tumor-bearing mice to tumor cells with different effector/target ratios (splenic lymphocytes∶Caki-1 cells or splenic lymphocytes∶YAC-1 cells). Results ODN1982 had no significant effect on in vitro proliferation of Caki-1 cells, while 500 μg/mL CpG ODN1826 significantly inhibited proliferation of Caki-1 cells. The tumor volumes of low-dose CpG ODN1826 group and high-dose CpG ODN1826 group were significantly smaller than those of control group and ODN1982 group at the end point of experiment (P<0.05), and there were significant differences in tumor weight among groups (P<0.05). It was revealed by histological observations that there were more inflammatory cell infiltration in low-dose CpG ODN1826 group and high-dose CpG ODN1826 group. The cytolytic activity of splenic lymphocytes of mice in high-dose CpG ODN1826 group to Caki-1 cells and YAC-1 cells was (9.74±1.16)% and (79.65±5.23)%, respectively at effector/target ratio of 50∶1, and was significantly higher than that in control group [(7.67±0.22)% and (10.12±3.03)%, respectively] and ODN1982 group [(7.66±0.93)% and (27.60±9.83)%, respectively] (P<0.05). Conclusion CpG ODN can inhibit renal carcinoma cell proliferation in vitro and tumor growth in tumor-bearing nude mice, and the mechanism may relate to the enhancement of innate immunity by CpG ODN.

Key words: CpG oligodeoxynucleotide, renal carcinoma, innate immunity