论文部分内容阅读
目的Ｂａｋ基因的过表达能够使ＨＣＣ－９２０４细胞的Ｇ１期延长并导致细胞凋亡，探讨ｐ２７ＫＩＰ１基因是否在这一过程中起重要作用。方法建立Ｂａｋ诱导ＨＣＣ—９２０４细胞凋亡和细胞周期停滞的模型。从Ｂａｋ诱导的凋亡模型中获得Ｐ２７ＫＩＰ１基因并测序。构建ＰＭＤ－ＫＩＰ１可诱导型表达系统并转染 ＨＣＣ－９２０４细胞，获得稳定转染子。 进一步研究ｐ２７ＫＩＰ１对细胞生长和细胞周期的调控作用。Ｗｅｓｔｅｒｎ ｂｌｏｔ证实在这一过程中该基因的表达水平上调。结果在诱导 ４８ ｈ后 ｐ２７ＫＩＰ１基因稳定转染细胞的活力降低 ３５％。在诱导后 ２４ｈ时 Ｐ２７ＫＩＰ１能够使 ＨＣＣ－９２０４细胞在Ｇ１期停滞，累积的细胞增加 ４０％。结论 Ｂａｋ使ＨＣＣ－９２０４细胞的 Ｇ１期延长是通过上调 ｐ２７ＫＩＰ１的表达；可诱导型 Ｐ２７ＫＩＰ１基因的过表达能够诱导 Ｇ１期停滞。
OBJECTIVE: Overexpression of Bak gene can prolong the G1 phase of HCC-9204 cells and lead to apoptosis, and investigate whether p27KIP1 plays an important role in this process. Methods The model of Bak induced HCC-9204 cell apoptosis and cell cycle arrest was established. The P27KIP1 gene was obtained from Bak-induced apoptosis model and sequenced. PMD-KIP1 inducible expression system was constructed and transfected into HCC-9204 cells to obtain stable transfectants. Further study of p27KIP1 on cell growth and cell cycle regulation. Western blot confirmed that the gene expression was up-regulated in this process. Results The viability of p27KIP1-stably transfected cells was reduced by 35% after 48 h of induction. At 24 h after induction, P27KIP1 was able to arrest HCC-9204 cells in G1 phase with a cumulative increase of 40% in cells. Conclusion Bak prolonged G1 phase of HCC-9204 cells by up-regulating the expression of p27KIP1. In overexpression of inducible P27KIP1 gene induced G1 arrest.