InnVit基因对BIOBR细胞增殖、凋亡及细胞周期的影响  

Effect of InnVit gene expression on proliferation, apoptosis and cell cycle of mouse melanocyte cell line BIOBR

作　　者：林福全[1] 关翠萍[2] 刘东银[2] 洪为松[2] 周妙妮[2] 许爱娥[2] 

机构地区：[1]浙江理工大学生物工程研究所,杭州310018 [2]安徽医科大学杭州市第三人民医院皮肤科

出　　处：《中华皮肤科杂志》2010年第4期269-272,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30671896,30872280）;浙江省自然科学基金（Z206607）

摘　　要：目的分析InnVit基因的抑制和过表达对永生化黑素细胞株BIOBR细胞增殖、周期及凋亡的影响，探讨InnVit基因的生物学功能及其对白癜风中黑素细胞缺失的影响。方法化学合成InnVit基因特异性siRNA片段以及构建过表达质粒载体P3XF—P120，lipofectamineTM2000脂质体方法转染B10BR细胞。半定量RT—PCR法检测InnVit基因mRNA水平；Western印迹检测该蛋白水平变化；M．rr法测定细胞增殖情况；流式细胞技术分析细胞周期和凋亡的变化。结果InnVit基因抑制后mRNA和蛋白表达水平明显低于对照组，InnVit基因过表达质粒组mRNA和蛋白表达水平显著高于对照组；转染后24、48、72h抑制组的细胞存活量极显著下降（P〈0．01），过表达质粒组的细胞存活量在48和72h极显著增加（P〈0．01）。转染后48h基因抑制组的早期凋亡率从（10．24±1．00）％升至（37．34±3．26）％，过表达质粒组早期凋亡率从（14．58±1．49）％降至（8．43±0．86）％，与对照组比较，差异均有统计学意义（P〈0．01）；转染48h基因抑制组的G。期细胞南（56．11±5．46）％增至（69．76±6．08）％（P〈0．05），过表达质粒组的G．期细胞由（55．14±5．65）％降至（29．33±3．01）％（P〈0．01）。结论InnVit基因抑制使细胞增殖能力下降，促进细胞凋亡；InnVit基因的过表达增强了细胞的增殖能力，抑制细胞凋亡。其增殖能力的改变可能是通过细胞周期的调控调节的。Objective To assess the effect of InnVit gene expression on cell proliferation, apoptosis and cell cycle of B10BR cells, and to explore the biological function of lnnVit gene and its influence on the loss of melanoeytes in vitiligo. Methods InnVit gene-targeting siRNA was synthesized, and overexpression plasmid P3XF-P120 was constructed： BIOBR cells were classified into 4 groups to be transfected with control siRNA （negative control）, siRNA targeting InnVit gene （siRNA group）, empty plasmid （plasmid control group）, and P3XF-P120 plasmid （P3XF-P120 group）, respectively. After additional culture for various periods, the mRNA and protein levels of InnVit gene were detected by RT-PCR and Western blot, respectively, cell survival by MTF assay, cell cycle and apoptosis by flow cytometry. Results The expression of InnVit mRNA and protein significantly increased in P3XF-P120 group compared with the plasmid control group, but decreased in siRNA group compared with the negative control group （all P 〈 0.01）. After the transfection, the survival rate of cells was significantly lower in siRNA group than in negative control group at 24, 48 and 72 hours, but higher in P3XF-P120 group than in plasmid control group at 48 and 72 hours （all P 〈 0.01 ）. At 48 hours after the transfection, the early apoptosis rate and proportion of cells in GI phase increased from （10.24 ± 1.00）% and （56.11 ± 5.46）% in negative control group to （37.34 ± 3.26）% and （69.76 ± 6.08）%, in siRNA group, respectively （P〈 0.01 or 0.05）, from （14.58 ± 1.49）% and （55.14 ± 5.65）% in plasmid control group to （8.43 ± 0.86）% and （29.33 ± 3.01 ）%, in P3XF-P120 group, respectively （both P 〈 0.01 ）. Conclusions The suppression of InnVit gene efficiently inhibits the proliferation and induces apoptosis of B10BR cells, and vice versa. Therefore, the regulation of cell cycle may affect the proliferation of B10BR cells.

关 键 词：InnVit基因 白癜风 细胞增殖 细胞凋亡 细胞周期 细胞 培养的 

分 类 号：R285[医药卫生—中药学]