摘要
目的:探讨缺氧诱导因子1α(HIF-1α)对直肠癌细胞增殖的影响,阐明其可能机制。方法:野生型直肠癌细胞系HT-29(Wt-HT-29)及转染HIF-1α特异性小干扰RNA(siRNA)的转染型HT-29细胞系(Si-HT-29)分别在常氧及缺氧状态下培养,通过Western blotting方法检测各组细胞在常氧及缺氧状态下HIF-1α和己糖激酶Ⅱ(HK-Ⅱ)的表达情况,采用MTT法检测各组细胞的生存率,采用ATP试剂盒检测细胞ATP生成量。结果:Western blotting结果,缺氧状态下野生型HT-29细胞中HIF-1α和HK-Ⅱ表达量高于转染型细胞。以常氧状态下野生型HT-29细胞的ATP生成量及细胞数量分别作为ATP及MTT数据的100%,将常氧状态下转染型HT-29细胞、缺氧状态下野生型及转染型HT-29细胞的ATP生成量及细胞数量与常氧状态下野生型HT-29细胞比值的百分数作为各组的ATP生成量及细胞生存率。常氧状态下转染型HT-29细胞的ATP生成量及细胞生存率分别为93.6%和92.0%,与野生型HT-29细胞比较差异无统计学意义(P>0.05)。在缺氧状态下野生型及转染型HT-29细胞的生存率分别为85.0%和61.3%,二者比较差异有统计学意义(P<0.05);同时在缺氧状态下野生型及转染型HT-29细胞ATP生成量分别为81.2%和42.9%,二者比较差异有统计学意义(P<0.05)。结论:HIF-1α能诱导直肠癌细胞表达HK-Ⅱ,增加ATP生成,从而发挥促增殖作用,HIF-1α可能成为直肠癌新的治疗靶点。
Objective To investigate the effect of hypoxia inducible transcription factor la （HIF-la） on proliferation of rectal cancer HT-29 cells and to clarify its mechanism. Methods The wild type HT-29 cells （Wt-HT-29） and Si-HT-29 cells transfected with HIF-la small interfering RNA were cultivated under normoxia and hypoxia conditions. The expressions of HIF-la and hexokinase-II （HK-II ） under normoxia and hypoxia conditions were detected by using Western blotting method. The survival rates of cells in various groups were detected by using MTT method. Furthermore, the ATP content was detected by using ATP Kit. Results Western blotting showed that the expressions of HIF-1α and HK-II in Wt-HT-29 cells were higher than those in Si-HT-29 cells under hypoxia condition. The amount of ATP and the number of wild-type cells generated under normoxic conditions were regarded as 100%. The percentages of the ratios of the amount of ATP and the number of Si-HT-29 cells under normoxia condition and the amount of ATP and the number of the two cell lines under hypoxia condition to the amount of ATP and the number of Wt-HT-29 cells under normoxia condition were regarded as the respective amount of ATP and the surrival rate of cells. Under normoxia condition the ATP amount and cell survival rate of HT-29 cels were 93.6% and 92.0%, respectively; and there was no significant difference compared with Wt-HT-29 cells （P〉 0.05）. But under hypoxia condition the survival rates of Wt-HT-29 cells and Si-NT-29 cells were 85.0G and 61.3M, respectively, and there was significant difference （P〈0.05）. At the same time the ATP amounts of Wt-HT-29 cells and Si-HT-29 cells were 81.2% and 42.9G, respectively; and significant difference was observed （P〈0.05）. Oonclusion HIF-1α can improve ATP production of rectal cancer cells by inducing HK-1α expression to increase the proliferation, and it may become a new therapy target of rectal cancer.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2013年第1期51-55,共5页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅科研基金资助课题(201101074)
关键词
缺氧诱导因子1Α
直肠肿瘤
腺苷三磷酸
缺氧
hypoxia inducible transcription factor la
rectal neoplasms
adenosine triphosphate
hypoxia
