氯化锰标记前列腺癌细胞株的体外MRI研究

摘要

Objective To assess the feasibility and security of prostate cancer cell lines (PC-3) laheled with manganese chloride (MnCl2) for magnetic resonance imaging(MRI) in vitro.Methods The PC-3 that purchased from American Type Culture Collection(ATCC) were recovered, cultured and amplified.The PC-3 were cultured in F-12 HAM'S medium with different concentrations of MnCl2 in cell incuhator and collected for MRI after 1 hour.The labeled cells were also collected for MRI in different amount and different time after labeling.The labeled cells were incubated with verapamil for 4 hours and the changes of the labeled cellular signal intensities were recorded in different time.Cell Counting Kit-8 (CCK-8) was used to determine the activities of the labeled cells.Results The PC-3 labeled with MnCl2 were high signalintensities on T1-weighted MRI.There were statistically significant differences between labeled cells and unlabeled cells (P ＜ 0.01).Highest signal intensity reached when PC-3 were labeled with 1.0 mM MnCl2.The signal intensity obviously decreased after 24 hours and became to normal signal intensity of unlabeled PC-3 after 72 hours.The PC-3 laheled with 1.0 mM MnCl2 solution showed high signal intensity on T1-weighted MRI with the minimum cell amount of 5.0 × 105 and lasted to 72 hours after a 4 hours incubation with verapamil.After 4 hours labeling, except the concentration of 0.1 mM.the other concentrations of MnCl2 ( ＞ 0.1mM ) had a certain toxicity on PC-3 (P ＜ 0.05).But, after 24 hours, the concentrations of 0.5-1.0 mM had little effect on the viability of PC-3 (P ＞ 0.05).Conclusion The PC-3 could he labeled with MnCl2 and appears high signal intensity on T1-weighted MRI.The PC-3 can be safety labeled with MnCl2 in concentrations which were equal or less than 1.0 mM.hut the duration of Mn+2 in PC-3 is shorter.Calcium channel blocker (verapamil) may be extend the duration of PC-3 laheled with MnCl2.%目的 初步探索使用氯化锰(MnCl2)标记前列腺癌细胞株(PC-3)行体外MRI的可行性和安全性.方法 将PC-3进行复苏、培养和扩增.在细胞培养箱中,PC-3与8组含不同MnCl2浓度的F-12 HAM'S培养基共同培养1h,收集已标记的PC-3并行MRI.另对不同细胞数量级和不同时间点的MnCl2标记的PC-3行MRI.用含维拉帕米的培养基培养MnCl2标记的PC-3 4 h,在不同时间点取标记后的PC-3行MRI.用WST-8法测定氯化锰标记的PC-3活性.结果 MnCl2标记的PC-3在T1WI显示为高信号,其信号强度与未标记的PC-3形成明显差异(P＜0.01),以1.0 mmol /L MnCl2为标记浓度时信号强度最强.1.0mmol/L MnCl2标记的PC-3在细胞数量为0.5×106时可呈高信号.在体外培养的条件下,标记后24 h的PC-3信号强度明显下降,标记后72 h基本恢复至未标记的PC-3信号强度水平.维拉帕米可延长MnCl2标记的PC-3有效成像时间至72 h.标记后4 h,除0.1 mmol/L MnCl2对PC-3的活性没有影响(P ＞ 0.05)外,其余各浓度组(＞ 0.1mmol/L)的MnCl2对PC-3的活性均有不同程度的影响(P ＜ 0.05);标记后24h,0.5 ～ 1.0 mmol/L MnCl2对细胞的活性影响不明显(P ＞ 0.05).结论 MnCl2可以有效标记PC-3,并能在T1WI以高信号成像且具有一定灵敏度.在浓度低于或等于1.0 mmol/L时,标记PC-3有一定的安全性,但标记维持时间较短.钙离子通道阻滞剂(维拉帕米)可适当延长PC-3的MnCl2标记维持时间.