国际肿瘤学杂志 ›› 2015, Vol. 42 ›› Issue (11): 801-804.doi: 10.3760/cma.j.issn.1673422X.2015.11.001

• 论著 •    下一篇

IL-32β提高宫颈癌C33A细胞对低氧低糖环境耐受的实验研究

孙树兰,郑晓霞,温莉,苏晋,何燕   

  1. 251700 山东省滨州市中心医院妇科(孙树兰、郑晓霞),眼科(温莉),骨科(苏晋),   放射科(何燕)
  • 出版日期:2015-11-08 发布日期:2015-09-30
  • 通讯作者: 郑晓霞,Email: 3060294653@qq.com E-mail:3060294653@qq.com

Study on enhanced resist to hypoxic/hypoglycemic condition by IL32β in cervical carcinoma C33A cells

Sun Shulan, Zheng Xiaoxia, Wen Li, Su Jin, He Yan.   

  1. Department of Gynecology, Binzhou City Center Hospital, Binzhou 251700, China
  • Online:2015-11-08 Published:2015-09-30
  • Contact: Zheng Xiaoxia E-mail:3060294653@qq.com

摘要: 目的探讨IL32β提高宫颈癌C33A细胞对低氧低糖环境耐受的作用机制。方法分别在正常条件和低氧低糖条件培养宫颈癌C33A细胞20 h后,使用实时定量PCR(RTPCR)和Western blotting检测IL32β的mRNA和蛋白水平。台酚蓝染色检测低氧低糖条件下培养的C33A细胞(对照组)以及添加10、100、500 ng/ml IL32β C33A细胞的生存率。用腹腔注射法构建裸鼠移植瘤,在分别注射0、1.0 mg/kg IL32β处理后测定移植瘤体积。使用siRNA构建IL32β沉默的细胞模型,检测VEGF的表达水平。结果低氧低糖条件下C33A细胞中的IL32β mRNA和蛋白水平分别是正常条件下的(6.12±0.03)倍和(2.23±0.04)倍(F=43.16,P<0.05;F=22.32,P<0.05)。低氧低糖条件下C33A细胞生存率为(51.92±3.41)%,而10、100、500 ng/ml IL32β组生存率分别为(55.23±3.92)%、(62.52±4.14)%、(69.14±2.45)%,与对照组比较差异均有统计学意义(F=14.25,P<0.05;F=35.53,P<0.01;F=56.28,P<0.01)。移植瘤培养28 d时,0 mg/kg IL32β组小鼠移植瘤体积为(578±64)mm3,而1.0 mg/kg组达(1 402±142)mm3(F=27.84,P<0.01)。此外,相对于正常C33A细胞,IL32β基因敲除的C33A细胞中VEGF的表达显著降低(F=36.85,P<0.05)。结论IL32β可提高C33A细胞对低氧低糖环境的耐受,其机制与增加VEGF表达水平相关。

关键词: 宫颈肿瘤, 白细胞介素类, 血管内皮生长因子类, 低氧低糖

Abstract: ObjectiveTo explore the enhancement effects and mechanisms of IL32β on human cervical carcinoma cells C33A to hypoxic/hypoglycemic condition. MethodsAfter cultured in hypoxia/hypoglycemic circumstance and normal circumstance for 20 hours respectively, the mRNA and protein expression of IL32β in C33A cells were detected by real timepolymerase chain reaction (RTPCR) and Western blotting respectively. Trypan blue stain was used to detect C33A cells viability in hypoxia/hypoglycemic circumstance and adding 10, 100, 500 ng/ml IL32β circumstance. The xenografted tumor of nude mice was established by intraperitoneal injection, and their volumes were tested for a given time after injecting 0, 1.0 mg/kg IL32β. siRNA was used to construct IL32β knockdown cells and detect the expression of VEGF. ResultsUnder the hypoxia/hypoglycemic circumstance, the expressions of IL32β mRNA were (6.12±0.03) times of the normal circumstance (F=43.16, P<0.05), the expressions of IL32β protein were (2.23±0.04) times of the normal circumstance (F=22.32, P<0.05). The C33A cells viability in hypoxia/hypoglycemic circumstance was (51.92±3.41)%, whereas, viability in 10 ng/ml IL32β group was (55.23±3.92)% (F=14.25, P<0.05), viability in 100 ng/ml IL32β group was (62.52±4.14)% (F=35.53, P<0.01), viability in 500 ng/ml IL32β group was (69.14±2.45)% (F=56.28, P<0.01). After 28 days, the volume of xenografted tumor of 0 mg/kg IL32β group was (578±64)mm3, and 1.0 mg/kg IL32β group up to (1 402±142)mm3 (F=27.84, P<0.01). In addition, compared with control group, the expression of VEGF in IL32β knockdown C33A cells was significantly decreased (F=36.85, P<0.05). ConclusionIL32β can enhance the resistance to hypoxic/hypoglycemic condition of C33A cells, which is associated with the increase of VEGF.

Key words: Uterine cervical neoplasms, Interleukins, Vascular endothelial growth factors, Hypoxic/hypoglycemic