摘要
目的研究量子点-RGD(QDs-RGD)作为荧光探针对胰腺癌细胞株SW1990靶向标记情况及生物相容性,探讨整合素靶向光动力学疗法(PDT)联合吉西他滨对SW1990细胞和胰腺癌裸鼠移植瘤的治疗效果。方法构建QDs-RGD整合素靶向探针。MTT法检测QDs-RGD探针对SW1990细胞的毒性,流式细胞术(FCM)检测SW1990细胞凋亡和细胞周期,RT-PCR检测SW1990细胞中髓样细胞白血病-1(Mcl-1)、丝氨酸/苏氨酸蛋白激酶B(Akt)和肿瘤坏死因子相关凋亡诱导配体(TRAIL)的表达,DCFH-DA探针检测活性氧水平。将胰腺癌荷瘤裸鼠模型分为空白对照组、单纯光辐照组、PDT组、吉西他滨组以及联合治疗组,测量各组瘤体积和瘤质量,计算抑瘤率。结果成功构建QDs-RGD整合素靶向探针。MTT检测显示,QDs-RGD探针浓度在≥20 nmol/L时对SW1990细胞增殖有抑制作用。FCM检测显示,PDT组细胞凋亡率为(17.86±1.23)%,高于空白对照组、单纯光辐照组和QDs-RGD探针组(P<0.01);且PDT组G_0/G_1期细胞比例最高,S期比例最低(P<0.01)。RT-PCR检测显示,PDT组Mcl-1、Akt mRNA水平(0.3394±0.0451,0.6161±0.0032)低于空白对照组、单纯光辐照组和QDs-RGD探针组,而TRAIL mRNA水平(0.7239±0.0017)高于其他3组(P<0.05)。PDT组荧光表达的SW1990细胞数为(286±6)个,活性氧水平高于其他3组(P<0.01)。联合治疗组瘤质量和瘤体积显著低于空白对照组、单纯光辐照组、PDT组和吉西他滨组(P<0.05);联合治疗组的抑瘤率为70.50%,高于吉西他滨组的43.53%和PDT组的37.05%(P<0.05)。结论QDs-RGD整合素靶向探针介导的PDT能抑制胰腺癌SW1990细胞增殖,促进细胞凋亡。以QDs-RGD作为光敏剂介导的PDT联合吉西他滨治疗能抑制胰腺癌裸鼠移植瘤的生长。
Objective To study the quantum dots synthesized RGD（ QDs-RGD） as a fluorescent probe targeted markers on pancreatic cancer cell line SW1990 and biocompatibility,and investigate the effect of integrin targeted photodynamic therapy（ PDT）combined with gemcitabine on pancreatic cancer cell line SW1990 and nude mice bearing pancreatic cancer xenograft. Methods QDsRGD integrin targeted probe was constructed. MTT method was used to detect the cytotoxicity of QDs-RGD probe on SW1990 cells.Flow cytometry（ FCM） was employed to test the apoptosis and cycle changes of SW1990 cells. RT-PCR was used to detect the mRNA levels of myeloid leukemia-1（ Mcl-1）,serine/threonine protein kinase B（ Akt） and tumor necrosis factor related apoptosis inducing ligand（ TRAIL） of SW1990 cells. Reactive oxygen species was tested by DCFH-DA probe. Nude mice bearing pancreatic cancer xenograft were divided into blank control group,radiation group,PDT group,gemcitabine group and combination group. The tumor volume and tumor weight were measured and tumor inhibitory rate was calculated. Results QDs-RGD integrin targeted probe was successfully constructed. MTT assay showed that the concentration of QDs-RGD probe ≥20 nmol/L could inhibit the proliferation of SW1990 cells.FCM assay showed that the cell apoptotic rate of PDT group was（ 17. 86±1. 23） %,higher than that of blank control group,radiation group and QDs-RGD probe group（ P〈0. 01）; Meanwhile,the ratio of SW1990 cells in G0/G1 phase was the highest,while that in S phase was the lowest（ P〈0. 01）. RT-PCR assay showed that the mRNA levels of Akt and Mcl-1（ 0. 3394± 0. 0451,0. 6161± 0. 0032）in PDT group were lower than those of blank control group,radiation group and QDs-RGD probe group（ P〈0. 05）,while that of TRAIL（ 0. 7239±0. 0017） was higher than the other 3 groups（ P〈0. 01）. The number of SW1990 cells expressed by fluorescence was286±6 in PDT group,which indicated that the reactive oxygen species level was higher than other groups（ P〈0. 01）. The tumor weight and tumor volume in combination group were lower than blank control group,radiation group,PDT group and gemcitabine group（ P〈0. 05）. The tumor inhibitory rate was 70. 50% in combination group,higher than 37. 05% in PDT group and 43. 53% in gemcitabine group（ P〈0. 05）. Conclusion QDs-RGD integrin targeted probe mediated PDT can significantly inhibit cell proliferation and promote cell apoptosis of pancreatic cancer cell line SW1990. QDs-RGD as photosensitizer mediated PDT combined with gemcitabine can significantly inhibit the growth of xenograft in nude mice.
作者
周敏
瞿春莹
徐雷鸣
金震东
ZHOU Min;QU Chunying;XU Leiming;JIN Zhendong(Department of Digest Endoscopic Diagnosis and Treatment,Xinhua Hospital Affiliated to Medical School of Shanghai Jiaotong University,Shanghai 200092,Chin)
出处
《临床肿瘤学杂志》
CAS
北大核心
2018年第4期289-297,共9页
Chinese Clinical Oncology
