摘要
目的:研究光动力疗法(photodynamic therapy,PDT)对人胆管癌细胞QBC939凋亡的影响.方法:体外培养人胆管癌QBC939细胞株,用不同浓度血卟啉衍生物(hematoporphyrin derivative,HPD)处理,并用半导体激光治疗仪不同强度光照后,采用CCK8法检测PDT对QBC939细胞的相对抑制率;应用流式细胞仪检测PDT作用前后QBC939细胞的凋亡率;RT-PCR检测PDT作用前后QBC939细胞中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)基因表达情况;SP免疫细胞化学法测定PDT作用前后QBC939细胞核中PCNA的表达情况.结果:CCK8检测结果显示PDT在体外能够抑制胆管癌QBC939细胞生长,8mg/LHPD经5J/cm2光照时实验组吸光度(A)值(0.403±0.027)与对照组A值(2.028±0.013)有统计学差异(P<0.05),细胞生长抑制率达80%,继续增加药物浓度或光照强度,细胞生长抑制率升高不明显,差异无统计学意义;流式细胞技术检测显示PDT能够明显促进胆管癌QBC939细胞的早期凋亡(74.6%±1.5%);RT-PCR检测显示PDT能够明显抑制胆管癌QBC939细胞中PCNA基因表达(0.68±0.06);SP免疫细胞化学结果显示PDT能够明显抑制胆管癌QBC939细胞核中PCNA蛋白表达(4.5%±1.4%).结论:PDT能够抑制胆管癌QBC939细胞生长,促进QBC939细胞的早期凋亡.PDT对QBC939细胞生长的抑制作用可能是通过抑制QBC939细胞的PCNA基因和蛋白水平的表达,从而促进其早期凋亡实现的.
AIM： To investigate the effect of photodynamic therapy on cholangiocarcinoma QBC939 cell apoptosis. METHODS： Cultured cholangiocarcinoma QBC939 cells were given different concentra- tions of hematoporphyrin derivatives and differ- ent light intensity. The relative growth inhibitionrate of QBC939 cells was detected by the CCK8 method. Flow cytometry assays were applied to determine the rate of QBC939 cell apoptosis. RT- PCR was used to detect transcriptional changes of proliferating cell nuclear antigen （PCNA）. Imrnunocytochemistry was used to measure the changes in protein expression of PCNA. RESULTS： PDT inhibited QBC939 cell growth in vitro, and significant different A values between the PDT group and control group was observed （0.403±0.027 vs 2.028 ±0.013, P 〈 0.05）. When the concentration of HPD was 8 mg/L and the light irradiation was 5 J/cm2, the relative growth inhibition rate of QBC939 cells was 80%. The increase in drug concentration or light dose did not significantly increase the growth inhibition rate. Flow cytometry assays showed that PDT could promote early apoptosis of QBC939 cells （74.6%±1.5%）. PDT could reduce the transcrip- tional changes of PCNA （0.68±0.06） and down- regulated the expression of PCNA protein in QBC939 cell nuclei （4.5% ±1.4%）. CONCLUSION： PDT could inhibit QBC939 cell growth and promote early apoptosis of QBC939 cells. PDT induces QBC939 cell growth inhibi- tion possibly by inhibiting PCNA gene and pro- tein expression and promoting early apoptosis.
出处
《世界华人消化杂志》
CAS
北大核心
2013年第19期1853-1858,共6页
World Chinese Journal of Digestology
关键词
光动力疗法
胆管癌
增殖细胞核抗原
凋亡
Photodynamic therapy
Cholangiocarci-noma
Proliferating cell nuclear antigen
Apoptosis
