Liang Liming, Bi Wenxiang, Chen Weiwen, Lin Yani, Tian Yuanyuan
Department of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan, 250012, China.
Lasers Med Sci. 2018 Feb;33(2):227-232. doi: 10.1007/s10103-017-2331-6. Epub 2018 Jan 6.
We combined pyropheophorbide-a methyl ester, photodynamic therapy (MPPa-PDT), 670 ± 10 nm, 4 mW/cm, 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.
我们将焦脱镁叶绿酸-a甲酯光动力疗法（MPPa-PDT），波长670±10nm，4mW/cm，与单纯疱疹病毒1型胸苷激酶/更昔洛韦（HSV1-TK/GCV）联合使用，以提高治疗效果。我们构建了HSV1-TK表达载体GV230-TK，并在荧光显微镜下观察到明亮的绿色荧光。这表明重组质粒已成功转染至PC-3M前列腺癌细胞中。由于PC-3M细胞中丰富的葡萄糖调节蛋白78（GRP78）启动子可导致HSV1-TK的活性表达，因此收集细胞蛋白进行蛋白质印迹法检测HSV1-TK的表达。在CCK-8检测中（n = 6），联合治疗组的细胞存活率约为10%，低于单纯MPPa-PDT组（23%）和单纯HSV1-TK/GCV组（35%）（t检验，P < 0.05）。使用流式细胞术测量细胞毒性；联合治疗组的凋亡率约为38%，高于单纯MPPa-PDT组（约22%）和单纯HSV1-TK/GCV组（约19%）。结果表明，两种治疗方法联合使用可有效提高对PC-3M细胞的杀伤作用。
