FNBP1参与HeLa细胞的形态控制与生长调控

FNBP1 is involved in morphology control and growth regulation in HeLa cells

目的研究FNBP1在HeLa细胞形态控制及生长调控过程中的作用。方法运用RT-PCR、Western blot法在mRNA和蛋白水平验证FNBP1在HeLa细胞中的表达;运用RT-PCR、Western blot法检测靶向siRNA干扰HeLa细胞内源FNBP1的表达情况,并于完全沉默和表达恢复2个时相点检测HeLa细胞在细胞形态、细胞周期等方面的变化。结果FNBP1在HeLa细胞中稳定表达;FNBP1表达沉默后,HeLa细胞形态发生纤维状转变;FNBP1表达恢复后,HeLa细胞形态恢复至上皮状;FNBP1表达沉默后,干扰组处于S期的细胞为30.36%,较正常组(25.45%)明显增多(P<0.05);而G2期干扰组细胞比例(9.28%)低于正常组(11.88%,P<0.05);HeLa细胞周期在S期出现阻滞。结论 FNBP1作为关键调控分子,为HeLa细胞的形态建成及维持所必需;FNBP1可能参与HeLa细胞周期调控相关过程。

Objective To investigate the role of formin-binding protein 1 （ FNBP1 ） in the morphology control and growth regulation in HeLa cells. Methods The expression of FNBP1 at mRNA and protein levels in HeLa cells was observed by RT-PCR and Western blotting respectively. After si-FNBP1 vector was transfected into HeLa cells, the expression of FNBP1 was detected after 96 （total silence） and 192 h （expression restored） by RT-PCR and Western blotting. The biological effects after silence of endogenetic FNBP1 in morphology and cell cycle were observed by HE stain and flow cytometry. Results FNBP1 was expressed steadily in HeLa cells. After silence of endogenetic FNBP1, the cellular morphology of HeLa cells changed into branched fibrous shape, and then restored to the normal shape as epithelial cells. Silence of FNBP1 resulted in 30.36% ceils arrested in S phase, significantly increased compared with those in the normal group （ 25.45 %, P 〈 0.05 ）, while those in G2 stage （9.28%） were lower than the normal group （11.88%, P 〈 0.05）. These changes were recovered when FNBP1 was restored to express. Conclusion FNBP1 plays an important role in the morphology control in HeLa cells, and also participates in the cell growth regulation.