Combination of MPPa-PDT and HSV1-TK/GCV gene therapy on prostate cancer
Combination of MPPa-PDT and HSV1-TK/GCV gene therapy on prostate cancer
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MPPa-PDT 联合 HSV1-TK/GCV 基因治疗前列腺癌
DOI：
10.1007/s10103-017-2331-6
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发表时间：
2018-02-01
影响因子：
2.1
通讯作者：
Tian, Yuanyuan
中科院分区：
文献类型：
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作者：
Liang, Liming;Bi, Wenxiang;Tian, Yuanyuan
我们将焦脱镁叶绿酸甲酯、光动力疗法（MPPa-PDT）、670 +/- 10 nm、4 mW/cm(2)与单纯疱疹病毒1型胸苷激酶/更昔洛韦（HSV1-TK/GCV）相结合，以提高治疗效果。构建HSV1-TK表达载体GV230-TK，在荧光显微镜下观察到亮绿色荧光。表明重组质粒成功转染PC-3M前列腺癌细胞。由于PC-3M细胞中丰富的葡萄糖调节蛋白78（GRP78）启动子可以引起HSV1-TK的活跃表达，收集细胞蛋白进行Western blot检测HSV1-TK的表达。 CCK-8检测（n=6）中，联合治疗组细胞存活率约为10%，低于单纯MPPa-PDT组（23%）和单纯HSV1-TK/GCV组（35%）（t检验，P < 0.05）。采用流式细胞术测定细胞毒性；联合治疗组细胞凋亡率约为38%，高于单纯MPPa-PDT组（约22%）和单纯HSV1-TK/GCV组（约19%）。结果表明，两种处理联合使用可有效提高对PC-3M细胞的杀细胞效果。
We combined pyropheophorbide-a methyl ester, photodynamic therapy (MPPa-PDT), 670 +/- 10 nm, 4 mW/cm(2), with herpes simplex virus type 1 thymidine kinase/ganciclovir (HSV1-TK/GCV) to improve the therapeutic effect. We built HSV1-TK expression vector GV230-TK and we observed a bright green fluorescence under fluorescence microscope. It indicated the recombinant plasmid was transfected into PC-3M prostate cancer cells successfully. As the abundant glucose-regulated protein 78 (GRP78) promoter in PC-3M cells can cause active expression of HSV1-TK, cell protein was collected for western blot to determine the expression of HSV1-TK. In CCK-8 assay (n = 6), the cell survival rate of combined treatment group was about 10%, less than that of pure MPPa-PDT group (23%) and pure HSV1-TK/GCV group (35%) (t test, P < 0.05). Flow cytometry was used to measure the cytotoxicity; the apoptosis rate of combined treatment group was about 38%, higher than that of pure MPPa-PDT group (about 22%) and pure HSV1-TK/GCV group (about 19%). The results showed that the combination of the two treatments can effectively improve the cytocidal effect in PC-3M cells.