新型分子探针近红外荧光标记Zn-DPA监测肿瘤疗效的研究

摘要

Objective To investigate the feasibility of a novel molecular probe of Zn-DPA-PSS794 to monitor the efficiency of doxorubicin to ovarian cancer and compare with Cy5. 5-annexin V. Methods Efficiency of doxorubicin to OVCAR-8 cells in vitro was measured by MTT assay and flow cytometry. The in vivo studies were performed on an OVCAR-8 xenograft tumor model. Mice were divided into a control group and a treatment group. Each group was divided into 2 subgroups, DPA and annexin V. In the treatment group, the mice were treated with doxorubicin for 2 doses. All mice were performed optical imaging by Zn-DPA-PSS794 or Cy5.5-annexin V, respectively and then sacrificed. The tumor was separated and stained by HE. The expression of caspase-3 protein wasmeasured by Western blot. Results The IC50 of doxorubicin to OVCAR-8 was 6 n,inol/L. The percentage of apoptosis and dead cells was 35% after doxorubicin treatment. In the optical image, photons accumulated in the tumor either by Zn-DPA-PSS794 or Cy 5. 5-annexin V in the treatment group. That was negative in the control group. The fluorescence intensity had significant difference between the 2 groups( P <0.001). The nuclei were big and stained with deep color after the cells were stained with HE. The caspase-3 expression was high in the treatment group, while it was low in the control group. Conclusion Zn-DPA-PSS794 as a probe used by optical imaging can monitor the efficiency of doxorubicin to OVCAR-8 xenograft tumor, which is similar to Cy5.5-annexin V.%目的:探讨新型分子探针近红外荧光标记Zn-DPA( Zn-DPA-PSS794)通过光学成像监测阿霉素治疗卵巢癌疗效的可行性,并与Cy5.5-annexin V光学成像效果进行比较.方法:MTT和流式细胞术分析阿霉素对卵巢癌细胞株OVCAR-8的杀伤作用.将OVCAR-8种植到裸鼠皮下,成瘤后分为对照组和治疗组,每组再分2个亚组,即DPA组和annexin V组.治疗组静脉注射阿霉素,2次后各组裸鼠分别进行Zn-DPA-PSS794光学显像和Cy5.5-annexin V光学显像,并进行定量分析.处死裸鼠,分离肿瘤,切片后进行HE染色,Western 印迹检测肿瘤组织中caspase-3蛋白表达水平.结果:阿霉素对OVCAR-8的IC50为6μmol/L,阿霉素能体外诱导OVCAR-8凋亡、坏死,总效率达35％.阿霉素治疗裸鼠移植瘤48 h后,Zn-DPA-PSS794光学成像和Cy5.5-annexin V光学成像均为阳性,而对照组2种光学成像结果均为阴性,对照组和治疗组肿瘤部位的荧光强度差异有统计学意义(P＜0.001).肿瘤组织HE染色示肿瘤细胞核大,深染.治疗组中caspase-3蛋白高表达,而对照组低表达.结论:Zn-DPA-PSS794光学成像能有效监测阿霉素治疗OVCAR-8荷瘤裸鼠的疗效,与Cy5.5-annexin V显像效果类似.