Chang C K, Tsai C Y, Lin L S, Jou S B, Liao S S, Cheng J T
Department of Surgery, Mackay Memorial Hospital, Taipei City, Taiwan.
Neurosci Lett. 1997 Aug 29;232(2):115-8. doi: 10.1016/s0304-3940(97)00588-0.
In an attempt to understand the change of superoxide dismutase (SOD) in tumor cells by hypoxia and hypoxia-normoxia exposure, the present study performed an in vitro investigation using rat glioma cell line in culture. Hypoxia was induced by an incubation with nitrogen gas for 15 h followed the normoxia exposure with air for 30 min. Activity of SOD in cytosolic and particulate of cells was determined by the reduction of nitroblue tetrazolium. Changes of mRNA for Cu,Zn-SOD or Mn-SOD were also characterized using Northern blotting analysis. Hypoxic stress decreased the activity of SOD, both Cu,Zn-SOD and Mn-SOD, in glioma cells. Expression of mRNA for SOD was elevated by hypoxic stress and the increase of mRNA level for Cu,Zn-SOD was more marked than that for Mn-SOD. In response to hypoxia-normoxia exposure, an increase of activity with a lower mRNA level for Mn-SOD was observed in glioma cells. However, changes of Cu,Zn-SOD both the activity and the level of mRNA were not found in glioma cells by hypoxia-normoxia. The obtained results suggest that the SOD in glioma cells can be activated to compensate the damage from free radicals during hypoxic stress.
为了了解缺氧及缺氧-复氧暴露对肿瘤细胞中超氧化物歧化酶(SOD)的影响,本研究使用培养的大鼠胶质瘤细胞系进行了体外研究。通过用氮气孵育15小时诱导缺氧,随后用空气进行30分钟的复氧暴露。通过氮蓝四唑还原法测定细胞胞质和颗粒部分中SOD的活性。还使用Northern印迹分析来表征铜锌超氧化物歧化酶(Cu,Zn-SOD)或锰超氧化物歧化酶(Mn-SOD)的mRNA变化。缺氧应激降低了胶质瘤细胞中SOD(包括Cu,Zn-SOD和Mn-SOD)的活性。缺氧应激使SOD的mRNA表达升高,且Cu,Zn-SOD的mRNA水平升高比Mn-SOD更明显。在缺氧-复氧暴露后,胶质瘤细胞中观察到Mn-SOD活性增加而mRNA水平降低。然而,缺氧-复氧并未使胶质瘤细胞中Cu,Zn-SOD的活性及mRNA水平发生变化。所得结果表明,胶质瘤细胞中的SOD可被激活以补偿缺氧应激期间自由基造成的损伤。