层流剪切力对悬浮培养红豆杉细胞HMGR酶基因转录的影响

Effects of Laminar Shear Stress on HMGR Gene Transcription in Suspension Cultured Taxus Cell

以HMGR酶基因为研究对象,采用实时荧光RT-PCR技术,研究了Couette式反应器中0．3 Pa层流剪切力作用后悬浮培养的对数期东北红豆杉细胞HMGR酶基因转录情况．结果显示,在对数生长时期,未经剪切处理的悬浮培养虹豆杉细胞HMGR基因转录水平呈增长趋势,而剪切处理后的悬浮培养红豆杉细胞HMGR基因转录水平则基本保持不变．同时,对相应细胞生物量的测定结果也显示出类似的变化．以上结果说明在对数生长时期施加一定强度的剪切力,将使细胞生长发生停滞,谊生长停滞现象可能与因剪切所引起的HMGR基因转录水平下降有一定相关性．

The exponential phase cells of Taxus cuspidata in suspension culture were treated by 0.3 Pa laminar shear stress in a Couette-type bioreactor in short term. Transcriptional activity of 3-hydroxy-3-methylgutaryl coenzyme A reductase （HMGR） gene was determined by fluorescent real time RT-PCT. Transcriptional activity of HMGR gene was determined by fluorescent real time RT-PCT. The results show that the level of HMGR gene transcription trends to rise in suspension cultured Taxus cell without shear treatment, whereas the level of HMGR gene transcription is invariable after shear treatment. The similar results are also obtained from the biomass in Taxus cells with treatment and no treatment respectively. All these indicate that laminar shear stress stagnates HMGR gene transcriptional activity and growth of Taxus cells in the exponential phase. The stasis of cells growth may be partially related to the level decrease of HMGR gene transcription caused by shear stress.