缺氧对宫颈癌细胞Hedgehog信号通路及肿瘤干细胞标志物表达的影响

发布时间：2018-05-12 16:06

  本文选题：Hh信号通路 + 缺氧　； 参考：《延边大学》2014年硕士论文

【摘要】：目的 缺氧条件下观察宫颈癌Hela中Hedgehog (Hh)信号通路相关因子和肿瘤干细胞标志物的表达及球形体形成能力。 方法 宫颈癌Hela细胞加CoC12分别培养24h和48h后,采用定量RT-PCR法和Western-Blot方法检测缺氧诱导因子(HIF1α和HIF2α)、Hh信号通路相关因子(smo、Gli-1)和肿瘤干细胞标志物(oct-4, sox2, Nanog)的表达；采用球形体形成实验检查Hela细胞的集落形成能力。 结果 宫颈癌He1a细胞的HIF1α与HIF2α的表达量,在缺氧24h开始增加,随着时间的延长,其表达量明显增加。Smo和Gli-1的mRNA量在缺氧48h开始增加；72h时Smo量是常氧组的5.4倍(p0.01,p0.01),Gli-1量是常氧组的123倍(p0.001,p0.001)。肿瘤干细胞(CSCs)标志物mRNA量在缺氧24h时开始增高,随着缺氧时间的延长,其表达量增加明显；72h时0ct4量是常氧组的78.8倍,Sox2量是常氧组的787.4倍,Nanog量是常氧组的689.8倍(p0.001,p0.001,p0.001)。缺氧组形成球形体的能力明显高于常氧组(p0.01)。 结论 在缺氧条件下Hela细胞中Hedgehog信号通路被激活,并且诱导Hela细胞具有肿瘤干细胞性。
[Abstract]:Purpose The expression of Hedgehog Hhh signal pathway and the expression of tumor stem cell markers and the ability of spheroid formation in Hela were observed under hypoxia condition. Method After Hela cells were cultured with CoC12 for 24 h and 48 h respectively, the expression of HIF1 伪 and HIF2 伪 Hh signaling pathway related factors (Gli-1) and tumor stem cell markers (oct-4, sox2, Nanog-1) were detected by quantitative RT-PCR and Western-Blot. The colony forming ability of Hela cells was examined by spheroid formation test. Result The expression of HIF1 伪 and HIF2 伪 in He1a cells of cervical cancer began to increase at 24 h of hypoxia. With the prolongation of time, the expression of mRNA in Smo and Gli-1 was significantly increased at 48 h after hypoxia, and the Smo level was 5.4 times that in normoxic group at 72 h after hypoxia, and the amount of Gli-1 was 123 times of that in normoxic group (p 0.001, p0.001, p0.001). The amount of mRNA, a tumor stem cell mRNA marker, began to increase at 24 h after hypoxia. With the prolongation of hypoxia time, the expression of 0ct4 was 78.8-fold higher than that of normoxic group at 72 h. The amount of 0ct4 was 787.4 times higher than that of normoxic group and 689.8 times of that of normoxic group (P 0.001p0.001p0.001p0.001). The ability to form spheroids in hypoxia group was significantly higher than that in normoxic group (P 0.01). Conclusion Under hypoxia condition, Hedgehog signaling pathway was activated in Hela cells, and Hela cells were induced to be tumor-stem cells.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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