摘要
目的通过观察顺铂对人骨肉瘤SaOS-2细胞核苷酸结合寡聚域1(NOD1)和NOD2表达的影响,探讨顺铂治疗骨肉瘤的作用机制。方法SaOS-2细胞加入顺铂,浓度依次为0μmol/L、5μmol/L、10μmol/L、20μmol/L,命名为S0组、S5组、S10组、S20组,培养24h、48h、72h,分别采用CCK8法、实时荧光定量聚合酶链式反应(PCR)法和免疫荧光法测定细胞生长活率、NOD1和NOD2的表达。结果在细胞培养48h、72h,S5组细胞存活率显著低于S0组(65.53%比100.00%;46.43%比100.00%,χ^2=8.64、73.97,均P<0.01)。在细胞培养24h、48h、72h,S10组、S20组细胞存活率显著低于S0组(80.60%比100.00%、42.94%比100.00%、27.90%比100.00%;62.54%比100.00%、33.09%比100.00%、22.95%比100.00%,χ^2=20.99、79.72、112.50;45.40、67.56、125.20,均P<0.01),且S20组骨肉瘤细胞存活率显著低于S5组(62.54%比93.78%、33.09%比65.53%、22.95%比46.43%,χ^2=28.47、21.78、11.71,均P<0.01)。S5组NOD1和NOD2mRNA的表达量在骨肉瘤细胞培养48h、72h显著高于培养24h且高于S0组[(3.64±0.44)比(4.47±1.22)比(1.79±0.44)比(1.00±0.00);(6.88±2.79)比(6.86±2.40)比(2.29±0.70)比(1.00±0.00),F=29.12、24.11,均P<0.01]。顺铂作用细胞48h和72h,NOD1蛋白表达较S0组有升高趋势。NOD1和NOD2mRNA的表达量呈现显著直线正相关(n=36,r=0.92,P<0.01)。结论顺铂对骨肉瘤细胞的功能有提升作用,呈现时间、浓度依赖性,顺铂可能是治疗骨肉瘤有效的药物,其机制可能是通过NOD1和NOD2途径。
Objective To investigate the effects of cisplatin on the expression of nucleotide-binding oligomerization domain-like receptor(NOD) 1 and 2 in human osteosarcoma SaOS-2 cell line, and to explore the mechanism of cisplatin in the treatment of human osteosarcoma. Methods CCK-8 assay, real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR) and immumofluorescence methods were used to determine the growth survival rate and expression levels of NOD1 and NOD2 in osteosarcoma SaOS-2 cell line treated with cisplatin (0, 5, 10, 20 μmol/L, named group S0, group S5, group S10, group S20, respectively) for 24, 48, 72 hours. Results After treatment with cisplatin for 48 h or 72 h, the growth survival rates of SaOS-2 cells were significantly decreased in group S5 than those in group S0(65.53% vs.100.00%;46.43% vs.100.00%, χ^2=8.64, 73.97, all P<0.01). Moreover, after treatment with cisplatin for 24 h, 48 h or 72 h, the growth survival rates of SaOS-2 cells were significantly decreased in group S10 or group S20 than those in group S0(80.60% vs.100.00%, 42.94 vs.100.00%, 27.90% vs.100.00%;62.54% vs.100.00%, 33.09% vs.100.00%, 22.95% vs.100.00%, χ^2=20.99, 79.72, 112.50;45.40, 67.56, 125.20, all P<0.01), and the growth survival rates were significantly lower in group S20 than those in group S5(62.54% vs.93.78%, 33.09% vs.65.53%, 22.95% vs.46.43%, χ^2=28.47, 21.78, 11.71, all P<0.01). The expression levels of NOD1 mRNA and NOD2 mRNA in group S5 were significantly increased at 48 h or 72 h than those at 24 h, and were higher than group S0 when treated with 5μmol/L cisplatin[(3.64±0.44) vs.(4.47±1.22) vs.(1.79±0.44) vs (1.00±0.00);(6.88±2.79) vs.(6.86±2.40) vs (2.29±0.70) vs.(1.00±0.00), F=29.12, 24.11, all P<0.01]. And the expression levels of NOD1 protein had an increased tendency after 48 h or 72 h treatment with 5μmol/L cisplatin.Furthermore, the expression level of NOD1 mRNA was positively correlated with NOD2 mRNA(n=36, r=0.92, P<0.01). Conclusion Cisplatin can elevate the function of osteosarcoma cell in a dose- and time-dependent manners, cisplatin may act as a efficient drug to cure osteosarcoma desease, which may be related to NOD1 and NOD2 signal pathway.
作者
林晓
罗冬娇
王恩智
Lin Xiao;Luo Dongjiao;Wang Enzhi(Department of Osteology ,the Integrated Western and Traditional Chinese Medicine Hospital of Taizhou ,Taizhou , Zhejiang 317500,China;Department of Preclinical Medicine,Qianjiang College of Hangzhou Normal University, Hangzhou ,Zhejiang 310012,China;Department of Clinical Laboratory,the Integrated Western and Traditional Chinese Medicine Hospital of Taizhou ,Taizhou ,Zhejiang 317500,China)
出处
《中国基层医药》
CAS
2019年第1期19-23,共5页
Chinese Journal of Primary Medicine and Pharmacy
基金
浙江省温岭市科技计划项目(2016C123).
关键词
骨肉瘤
肿瘤细胞
培养的
顺铂
聚合酶链反应
核苷酸类
衔接蛋白质类
信号转导
药物评价
Osteosarcoma
Tumor cells, cultured
Cisplatin
Polymerase chain reaction
Nucleotides
Adaptor proteins, signal transducing
Drug fvaluation