99mTc-sestamibi to monitor treatment with antisense oligodeoxynucleotide complementary to MRP mRNA in human breast cancer cells
99mTc-sestamibi to monitor treatment with antisense oligodeoxynucleotide complementary to MRP mRNA in human breast cancer cells
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DOI：
10.1007/bf02985587
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发表时间：
2006-01-01
影响因子：
2.6
通讯作者：
Tonami, N
中科院分区：
文献类型：
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作者：
Kinuya, S;Bai, JM;Tonami, N
目的：利用tech -99m sestamibi （MIBI）评价恶性肿瘤的多药耐药（MDR）现象并预测其化疗效果。目前的研究探讨了在反义寡脱氧核苷酸（AS-ODN）治疗后，涉及Tc-99m-MIBI的多药耐药相关蛋白（MRP） mRNA表达监测变化的可能性。方法：采用人乳腺癌MCF-7细胞株及其耐多药诱导的MCF-7/VP细胞株。将两株细胞系1 × 10(4)个/ml的细胞悬液接种于24孔板（0.2 ml/孔），孵育1天。与MRP mRNA编码区互补的反义（AS） 20-mer硫代ODN及其义(S) ODN以最终浓度高达25 μ M给予，然后孵育5天。每孔加入Tc-99m-MIBI溶液，孵育30分钟。细胞Tc-99m-MIBI摄取校正蛋白浓度。通过逆转录聚合酶链反应（RT-PCR）分析MRP mRNA表达水平。结果：Tc-99m-MIBI在MCF-7/VP细胞中的细胞摄取仅为MCF-7细胞的15%。在25 μ M的AS-ODN处理5天后，MCF-7/VP细胞中的Tc-99m-MIBI摄取比未处理的对照细胞增加了2.4倍。MCF-7细胞对Tc-99m-MIBI的摄取不受AS-ODN的影响。Sense ODN不改变两种细胞系的摄取。RT-PCR证实AS-ODN处理后MCF-7/VP细胞中MRP mRNA减少。结论：AS-ODN给药对MRP功能的影响可以通过评估Tc-99m-MIBI的细胞摄取来监测。
Objective: Technetium-99m sestamibi (MIBI) has been utilized to evaluate multi-drug resistance (MDR) phenomenon of malignant tumors and to predict chemotherapeutic effects on them. The current investigation examined the possibility of monitoring changes with respect to mRNA expression of multi-drug resistance associated protein (MRP) following antisense oligodeoxynucleotide (AS-ODN) treatment involving Tc-99m-MIBI. Methods: The human breast cancer MCF-7 cell line and its MDR-induced MCF-7/VP cell line were employed. Cell suspensions of the two cell lines at I X 10(4) cells/ml were inoculated in 24-well plates (0.2 ml/well) and incubated for one day. Antisense (AS) 20-mer phosphorothioate ODN complementary to the coding region of MRP mRNA and its sense (S) ODN were administered at final concentrations up to 25 mu M, followed by a 5-day incubation. Tc-99m-MIBI solution was added to each well and incubated for 30 min. Cellular Tc-99m-MIBI uptake was corrected for protein concentration. MRP mRNA expression levels were analyzed via the reverse transcription polymerase chain reaction (RT-PCR). Results: Cellular uptake of Tc-99m-MIBI in MCF-7/VP cells was only 15% of that of MCF-7 cells. Following AS-ODN treatment at 25 mu M for five days, Tc-99m-MIBI uptake in MCF-7/VP cells increased 2.4-fold in comparison with non-treated control cells. Tc-99m-MIBI uptake in MCF-7 cells was unaffected by AS-ODN administration. Sense ODN did not alter uptake in either cell line. RT-PCR confirmed reduction of MRP mRNA in MCF-7/VP cells following AS-ODN treatment. Conclusion: Effects of AS-ODN administration on MRP function can be monitored via assessment of cellular uptake of Tc-99m-MIBI.