Influence of Photodynamic Therapy on Apoptosis and Invasion of Human Cholangiocarcinoma QBC939 Cell Line

Yun-jie Chen; Hai-tao Jiang; Jing-yu Cao

摘要

Objective To investigate the effect of photodynamic therapy (PDT) mediated by hematoporphyrin derivative (HPD) on apoptosis and invasion of cholangiocarcinoma QBC939 cell lines. MethodsIn vitrocultured cholangiocarcinoma QBC939 cell line was exposed to 2, 4, 6, 8, 10, 12, and 14μg/ml HPD with 5, 10, and 15 J/cm2 light intensity, respectively. The optical density at 450 nm of the QBC939 cells was measured by CCK8 assay and its growth inhibition ratio was calculated. Flow cytometry and transwell migration assay were applied to detect cell apoptosis and invasion respectively. RT-PCR and immunocytochemistry analyses were used to detect expressions of vascular endothelial growth factor-C (VEGF-C), cyclooxygenase-2 (COX-2), and proliferating cell nuclear antigen (PCNA). Enzyme-linked immunosorbent assay (ELISA) was carried out to examine the secretion of VEGF-C and COX-2 in QBC939 cells. Results Exposure to HPD-PDT can significantly suppress the growth of QBC939 cells (allP<0.05). HPD-PDT can promote apoptosis of QBC939 cells at the early stage. When the concentration of HPD was 2μg/ml and light irradiation was 5 J/cm2, HPD-PDT had no obvious inhibitory effect on QBC939 cell growth, but can obviously inhibit cell invasion, and significant difference was observed between the HPD-PDT and control groups (P<0.01). The HPD-PDT can reduce the mRNA and protein expressions of VEGF-C, COX-2, and PCNA, and decrease the secretion of VEGF-C and COX-2 in QBC939 cells. Conclusion PDT could promote apoptosis and inhibit growth and invasion of cholangiocarcinoma cells QBC939in vitro.

机译：目的探讨血卟啉衍生物（HPD）介导的光动力疗法（PDT）对胆管癌QBC939细胞凋亡和侵袭的影响。方法将体外培养的胆管癌QBC939细胞系分别以2、5、10和15 J / cm2的光强度暴露于2、4、6、8、10、12和14μg/ ml HPD。通过CCK8测定法测量QBC939细胞在450nm的光密度，并计算其生长抑制率。流式细胞术和transwell迁移法分别用于检测细胞凋亡和侵袭。使用RT-PCR和免疫细胞化学分析来检测血管内皮生长因子-C（VEGF-C），环氧合酶-2（COX-2）和增殖细胞核抗原（PCNA）的表达。进行酶联免疫吸附试验（ELISA）以检查QBC939细胞中VEGF-C和COX-2的分泌。结果暴露于HPD-PDT可显着抑制QBC939细胞的生长（allP <0.05）。 HPD-PDT可以在早期促进QBC939细胞凋亡。当HPD浓度为2μg/ ml，光照度为5 J / cm2时，HPD-PDT对QBC939细胞的生长没有明显的抑制作用，但可以明显抑制细胞的侵袭，与对照组相比，HPD-PDT有显着差异。组（P <0.01）。 HPD-PDT可以降低QBC939细胞中VEGF-C，COX-2和PCNA的mRNA和蛋白表达，并减少VEGF-C和COX-2的分泌。结论PDT在体外可促进胆管癌细胞QBC939的凋亡并抑制其生长和侵袭。

Influence of Photodynamic Therapy on Apoptosis and Invasion of Human Cholangiocarcinoma QBC939 Cell Line

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