Wang Fang, Chen Tong-Sheng, Xing Da, Wang Jin-Jun, Wu Yun-Xia
Institute of Laser Life Science, South China Normal University, Guangzhou 510631, China.
Lasers Surg Med. 2005 Jan;36(1):2-7. doi: 10.1002/lsm.20130.
Low-power laser irradiation (LPLI) has been used for therapies such as curing spinal cord injury, healing wound etc. Yet, the mechanism of LPLI remains unclear. In order to determine the effects of high fluence LPLI on cell growth and caspase-3 activity, we have measured the dynamics of caspase-3 activity during cell apoptosis induced by high fluence LPLI treatment.
STUDY DESIGN/MATERIALS AND METHODS: He-Ne laser was used to irradiate human lung adenocarcinoma cells (ASTC-a-1). Cell Counting Kit-8 was used for cytotoxicity assay. A fluorescent microscope was used to perform fluorescence resonance energy transfer (FRET) imaging. A luminescence spectrometer was used to acquire the fluorescent emission spectrum. Statistical analysis was performed with Student's paired t-test.
Cytotoxicity assay showed that when light irradiation fluence exceeded 60 J/cm2, LPLI treatment induced ASTC-a-1 cell apoptosis in a fluence-dependent manner. FRET imaging and spectrofluorometric analysis demonstrated that caspase-3 was activated during high fluence LPLI-induced cell apoptosis.
Using FRET technique, we have reported that high fluence LPLI can induce human lung adenocarcinoma cells (ASTC-a-1) apoptosis. The activation of caspase-3 plays an important role in the apoptotic process.
低功率激光照射(LPLI)已用于诸如治疗脊髓损伤、伤口愈合等疗法。然而,LPLI的机制仍不清楚。为了确定高能量密度LPLI对细胞生长和半胱天冬酶-3活性的影响,我们测量了高能量密度LPLI处理诱导细胞凋亡过程中半胱天冬酶-3活性的动态变化。
研究设计/材料与方法:使用氦氖激光照射人肺腺癌细胞(ASTC-a-1)。使用细胞计数试剂盒-8进行细胞毒性测定。使用荧光显微镜进行荧光共振能量转移(FRET)成像。使用发光光谱仪获取荧光发射光谱。采用学生配对t检验进行统计分析。
细胞毒性测定表明,当光照射能量密度超过60 J/cm2时,LPLI处理以能量密度依赖的方式诱导ASTC-a-1细胞凋亡。FRET成像和荧光光谱分析表明,在高能量密度LPLI诱导的细胞凋亡过程中半胱天冬酶-3被激活。
我们利用FRET技术报道了高能量密度LPLI可诱导人肺腺癌细胞(ASTC-a-1)凋亡。半胱天冬酶-3的激活在凋亡过程中起重要作用。