Feng Yi, Tian Zhong-Min, Wan Ming-Xi, Zheng Zhao-Bin
Department of Biomedical Engineering, Xi'an Jiaotong University, Xi'an 710049, Shaanxi Province, China.
World J Gastroenterol. 2008 Aug 21;14(31):4873-9. doi: 10.3748/wjg.14.4873.
To investigate the low intensity ultrasound (US)-induced apoptosis in human gastric carcinoma cells and its potential mechanism and to suggest a new therapeutic approach to gastric carcinoma.
Human SGC-7901 gastric carcinoma cells were cultured in vitro and irradiated by low intensity US for 10 min at different intensities with different incubation times after irradiation. Morphologic changes were examined under microscope with trypan blue staining and then the percentage of early apoptotic cells was detected by flow cytometry (FCM) with double staining of fluorescein isothiocyanate (FITC)-Annexin V/propidium iodide (PI). Two-dimensional electrophoresis (2DE) was used to get the protein profile and some proteins differently expressed after US irradiation were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Functional analysis was performed to investigate the mechanism of US-induced cell apoptosis.
The percentage of apoptotic cells increased about 10% after US irradiation (12.0 W/cm(2), 12 h culture). The percentage of early apoptosis and secondary necrosis in the US-irradiated cells increased with the increased US intensity. Moreover, apoptotic cells increased with the increased culture time after US irradiation and reached its maximum at about 12 h. Several new proteins appeared after US irradiation and were up or down regulated more than 2 times. Some heat shock proteins (HSPs) were found to be associated with the signal process simulating the apoptosis of cells.
Low intensity US could induce apoptosis in human gastric carcinoma cells. US-induced apoptosis is related to US intensity/culture time. US-induced apoptosis may be caspases-dependent and endoplasmic reticulum (ER) stress-triggered apoptosis may also contribute to it. Proteomic experimental system is useful in finding the protein alteration in carcinoma cells after US irradiation, helping to develop a new cancer therapy.
研究低强度超声诱导人胃癌细胞凋亡及其潜在机制,并提出一种新的胃癌治疗方法。
体外培养人SGC-7901胃癌细胞,用不同强度的低强度超声照射10分钟,照射后培养不同时间。用台盼蓝染色在显微镜下观察形态变化,然后用异硫氰酸荧光素(FITC)-膜联蛋白V/碘化丙啶(PI)双染法通过流式细胞术(FCM)检测早期凋亡细胞百分比。采用双向电泳(2DE)获取蛋白质图谱,并用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)鉴定超声照射后差异表达的一些蛋白质。进行功能分析以研究超声诱导细胞凋亡的机制。
超声照射(12.0W/cm²,培养12小时)后凋亡细胞百分比增加约10%。超声照射细胞中早期凋亡和继发性坏死的百分比随超声强度增加而增加。此外,超声照射后凋亡细胞随培养时间增加而增加,并在约12小时达到最大值。超声照射后出现几种新蛋白质,其上调或下调超过2倍。发现一些热休克蛋白(HSPs)与模拟细胞凋亡的信号过程有关。
低强度超声可诱导人胃癌细胞凋亡。超声诱导的凋亡与超声强度/培养时间有关。超声诱导的凋亡可能依赖于半胱天冬酶,内质网(ER)应激触发的凋亡也可能起作用。蛋白质组学实验系统有助于发现超声照射后癌细胞中的蛋白质变化,有助于开发新的癌症治疗方法。