90Y-DOTATOC对人胰腺神经内分泌瘤BON-1细胞的抑制作用

目的 探讨90Y-1,4,7,10-四氮杂环十二烷-1,4,7,10-四乙酸-酪氨酸3-奥曲肽（DOTATOC）对人胰腺神经内分泌瘤细胞BON-1的抑制作用。 方法 （1）采用90Y对DOTATOC进行标记，然后纯化90Y-DOTATOC；（2）考察90Y-DOTATOC的体外稳定性；（3）采用噻唑蓝（MTT）法，分别考察90Y、DOTATOC、90Y-DOTATOC对人胰腺神经内分泌瘤细胞BON-1和人胰腺胆管瘤细胞PANC-1的抑制作用，并分别计算细胞增殖抑制率。设立阴性对照组、阳性对照组（长春新碱，50 μmol/L）、DOTATOC组（25 μmol/L）、90Y组（1.8 MBq/ml）、90Y-DOTATOC高剂量组（90Y的放射性浓度为1.8 MBq/ml，DOTATOC的浓度为25 μmol/L）、90Y-DOTATOC中剂量组（90Y的放射性浓度为0.37 MBq/ml，DOTATOC的浓度为25 μmol/L）、90Y-DOTATOC低剂量组（90Y的放射性浓度为0.074 MBq/ml，DOTATOC的浓度为25 μmol/L）。组间比较采用独立样本t检验。 结果 （1）90Y标记DOTATOC的标记率为（61.93±3.53）%，放射化学纯度为（98.88±0.38）%，放射性浓度为4.6 MBq/ml，比活度为1.6 GBq/μmol。（2）90Y-DOTATOC在生理盐水和10%胎牛血清中放置7 d后的放射化学纯度分别为97.73%和97.02%。（3）加药24、48 h后，与阴性对照组相比，DOTATOC组对BON-1细胞的抑制作用均显著增高，且差异有统计学意义（t=2.654，3.981，均P<0.05）；加药24、48 h后，90Y-DOTATOC高、中剂量组对BON-1和PANC-1细胞的抑制作用均优于DOTATOC组，且差异有统计学意义（t=2.267~3.852，均P<0.05）；加药24、48 h后，90Y组对PANC-1和BON-1细胞的抑制作用均明显低于90Y-DOTATOC高剂量组，且差异有统计学意义（t=2.698~3.180，均P<0.05）。 结论 90Y-DOTATOC对BON-1细胞有较强的抑制作用，且与90Y的活度呈剂量依赖关系。与单独使用DOTATOC或者90Y对比，90Y-DOTATOC对BON-1细胞的抑制作用更强。

Inhibitory effect of 90Y-DOTATOC on human pancreatic neuroendocrine tumor cell BON-1

Objective To investigate the inhibitory effect of 90Y-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-Tyr (3)-octreotide (DOTATOC) on the human pancreatic neuroendocrine tumor cell line BON-1. Methods (1) DOTATOC was labeled with 90Y, and then purified it. (2) The stability of 90Y-DOTATOC was investigated in vitro. (3) The thiazole blue method was used to examine the inhibitory effects of 90Y, DOTATOC, and 90Y-DOTATOC on human pancreatic neuroendocrine tumor cell BON-1 and human pancreatic cholangioma cell PANC-1, and then the cell proliferation inhibition rate was calculated. Negative control group, positive control group (Vincristine, 50 μmol/L), DOTATOC group (25 μmol/L), 90Y group (1.8 MBq/ml), 90Y-DOTATOC high-dose group (90Y dose was 1.8 MBq/ml, DOTATOC concentration was 25 μmol/L), 90Y-DOTATOC medium dose group (90Y dose was 0.37 MBq/ml, DOTATOC concentration was 25 μmol/L), 90Y-DOTATOC low-dose group (90Y dose was 0.074 MBq/ml, DOTATOC concentration was 25 μmol/L) were set up. Independent sample t-test was used for inter group comparison. Result (1) The labeling rate of the DOTATOC labeled with 90Y was (61.93±3.53)%. Its radiochemical purity was (98.88±0.38)%, its radioactive concentration was 4.6 MBq/ml, and its specific activity was 1.6 GBq/μmol. (2) The radiochemical purity of 90Y-DOTATOC after being placed in physiological saline and 10% bovine serum for 7 days was 97.73% and 97.02%, respectively. (3) After 24 h and 48 h of drug addition, the inhibitory effect of the DOTATOC group on BON-1 cells was significantly increased compared with that of the negative group, and the difference was statistically significant (t=2.654, 3.981, all P<0.05). After 24 h and 48 h of drug addition, the inhibitory effects of the high and medium 90Y-DOTATOC dose groups on BON-1 and PANC-1 cells were superior to that of the DOTATOC group, and the difference was statistically significant (t=2.267–3.852, all P<0.05). After 24 h and 48 h of drug addition, the inhibitory effects of the pure 90Y group on PANC-1 and BON-1 cells were significantly lower than that of the high-dose 90Y-DOTATOC group, and the difference was statistically significant (t=2.698–3.180, all P<0.05). Conclusion 90Y-DOTATOC exerts a strong inhibitory effect on BON-1 tumor cells and is dose-dependent on 90Y dosage. Compared with using DOTATOC or 90Y alone, 90Y-DOTATOC exhibits better inhibitory effect on BON-1 tumor cells.