FNBP1参与HeLa细胞的形态控制与生长调控.docVIP

FNBP1参与HeLa细胞的形态控制与生长调控 李 鑫1,2，温 泉1,2，周云飞1,2，何玉霞1,2，张 军2 （400016重庆，重庆医科大学：分子医学与肿瘤研究中心1，细胞生物学及遗传学教研室2） ［摘要］ 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两个时相点检测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(P0.05)；而G2期干扰组细胞比例(9.28%)低于正常组(11.88%，P0.05)；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。 ［关键词］ FNBP1；HeLa细胞；形态控制；生长调控 [中图法分类号] [文献标志码] A FNBP1 is involved in morphology control and growth regulation of HeLa cells Li Xin，Wen Quan，Zhou Yunfei，He Yuxia，Zhang Jun (1 Molercular Medicine and Cancer Research Center, 2Department of Cell Biology and Genetics,Chongqing Medical University, Chongqing, 400016,China) [Abstract] Objective To investigate the role of FNBP1 in morphology control and growth regulation of HeLa cells. Methods The expression of FNBP1 in HeLa cells was observed by RT-PCR and Western blot respectively at mRNA and protein levels; si-FNBP1 molecules was transfected into HeLa cells, and the expression of FNBP1 was detected after 96 h and 192 h by RT-PCR and Western blot; The biological effects after silence of endogenetic FNBP1 were observed by HE stain and FCM. Results FNBP1 is expressed steadily in HeLa cells. After silence of endogenetic FNBP1, the cellular morphology of HeLa cells change in to branched fibrous shape and the blocking effect is exhibited in the S phase of the cell cycle:cells of the silence group is 30.36%, significantly increased compared with the normal group (25.45%, P0.05), while that in G2 stage (9.28%) is lower than the normal group (11.88%, P0.05). And these changes are recovered when FNBP1 is reexpressed. Conclusion FNBP1 plays a important role in morphology control and is involved in growth regulation of HeLa cells. [Key words] FNBP1; HeLa cells; morphology c