腺病毒介导的p16~(INK4a)基因沉默对人毛乳头细胞的影响
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摘要
目的利用RNA干扰技术探讨p16~(INK4a)对人毛乳头细胞(dermal papilla cell,DPC)的促进作用。方法将针对p16~(INK4a)基因设计的RNA腺病毒转染高传代DPC,观察细胞生长活性,电镜下观察细胞形态,western blot检测衰老相关蛋白β-半乳糖苷酶、pRb的表达。结果转染后细胞倍增时间缩短,细胞核分裂增多,高传代DPC中β-半乳糖苷酶、pRb蛋白较低传代DPC均上调,转染后两种蛋白的表达呈下降趋势。结论高传代DPC非凝集性生长与细胞过早性衰老有关,DPC的p16~(INK4a)基因沉默后可延缓DPC衰老,促进DPC增殖。
Objective Study the effect of p16~(INK4a) aon dermal papilla cells(DPCs) by RNA interference technique.Methods High passage DPCs in vitro were transfected by RNA adenovirus designed according to p16~(INK4a) agene sequence to observe the cell growth activity,and the cell morphology was observed by electron microscope.The expression of senescence related protein was detected by western blot.Results The cells doubling time of transfected group was shortened and the cell division was increased compared with normal group.The expression of pRb and β-galactosidase protein in high passage DPCs was up-regulated compared with low passage,but decreased after transfection.Conclusion Non-aggregative growth of DPCs is associated with premature aging.Knockdown of p16~(INK4a) ain DPCs can delay DPCs senescence and promote DPCs proliferation.
Objective Study the effect of p16~(INK4a) aon dermal papilla cells(DPCs) by RNA interference technique.Methods High passage DPCs in vitro were transfected by RNA adenovirus designed according to p16~(INK4a) agene sequence to observe the cell growth activity,and the cell morphology was observed by electron microscope.The expression of senescence related protein was detected by western blot.Results The cells doubling time of transfected group was shortened and the cell division was increased compared with normal group.The expression of pRb and β-galactosidase protein in high passage DPCs was up-regulated compared with low passage,but decreased after transfection.Conclusion Non-aggregative growth of DPCs is associated with premature aging.Knockdown of p16~(INK4a) ain DPCs can delay DPCs senescence and promote DPCs proliferation.
引文
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[2]Serrano M,Hannon GJ,Beach D.A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4[J].Nature,2016,366(6456):704-7.
[3]温进坤,韩梅.医学分子与生物学理论与研究技术[M].2版.北京:科学出版社,2002,365-366.
[4]Hall BM,Balan V,Gleiberman AS,et al.Aging of mice is associated with p16(Ink4a)-andβ-galactosidase-positive macrophage accumulation that can be induced in young mice by senescent cells[J].Aging,2016,8(9):1294-1311.
[5]Baker DJ,Wijshake T,Tchkonia T,et al.Clearance of p16Ink4a-positive senescent cells delays ageing-associated disorders[J].Nature,2011,479(7372):232-236.
[6]Jahoda CA,Reynolds AJ,Oliver RF.Induction of hair growth in ear wounds by cultured dermal papilla cells[J].Invest Dermatol,1993,101(4):584-590.
[7]吕中法,蔡绥,伍津津,等.培养毛乳头细胞生物学特性和毛囊重建的研究[J].中华皮肤科杂志,2006,39(2):64-67.
[8]刘官智,伍津津.永生化人毛乳头细胞系体内诱导毛囊的形成[J].中国皮肤性病学杂志,2011,25(3):177-180.
[9]陈欣玥,何黎,刘玲,等.毛囊混合细胞诱导毛囊重建的研究[J].中国皮肤性病学杂志,2011,25(1):15-17.
[10]董巍.细胞衰老与肿瘤[J].实用肿瘤杂志,2013,27(2):185-189.