Rapid Propagation of Virus-free Sugarcane Plantlets via Temporary Immersion Bioreactor System

By employing temporary immersion bioreactor system(TIBs),we studied virus-free culture of seedlings from sugarcane varieties ROC16 and ROC22,from medium recipe,inoculation amount,sucrose concentration,and variety difference. The results showed,using this method,that proliferation rate of ROC16 improved by 40 times,per flask generated about 800 plantlets; of ROC22 improved by 30 times,per flask generated about 400-600 plantlets. The results provided basis for using TIBs in rapid propagation of plantlets via tissue culture.

CAAP1对人肝癌细胞SMMC-7721增殖、迁移和侵袭的影响

目的探讨Caspase活性和凋亡抑制因子1(CAAP1)对人肝癌细胞SMMC-7721增殖、迁移和侵袭的影响。方法构建CAAP1的过表达载体pcDNA3/CAAP1和敲降载体pSilencer 2.1-U6 neo/shR-CAAP1(shR-CAAP1),并进行鉴定。SMMC-7721分为4组:pcDNA3/CAAP1组、pcDNA3对照组、shR-CAAP1组和pSilencer对照组。SMMC-7721培养后,将CAAP1的过表达载体pcDNA3/CAAP1(pcDNA3/CAAP1组)和敲降载体shR-CAAP1(shR-CAAP1组)以及它们的对照(pcDNA3对照组、pSilencer对照组)分别转染到SMMC-7721中,48 h后进行后续实验。实时定量PCR(qRT-PCR)检测各组细胞中CAAP1 mRNA的表达水平,Western blot检测CAAP1、cleaved Caspase-3蛋白的表达水平;CCK-8实验检测各组细胞的增殖,细胞集落形成实验比较各组细胞集落形成能力;划痕实验比较各组细胞运动能力,Transwell小室检测各组细胞的迁移和侵袭;流式细胞术检测各组细胞的凋亡。纳入TCGA数据库CAAP1低表达的患者75例、CAAP1高表达的患者295例随访48个月的数据,Kaplan-Meier生存曲线比较不同CAAP1表达水平对肝癌患者总生存(OS)的影响。结果双酶切鉴定表明CAAP1过表达载体pcDNA3/CAAP1和敲降载体shR-CAAP1构建成功;qRT-PCR和Western blot结果提示pcDNA3/CAAP1能够使SMMC-7721细胞中CAAP1 mRNA和蛋白表达水平增加(与pcDNA3对照组相比,P<0.05),而shRCAAP1使CAAP1 mRNA和蛋白表达水平降低(与pSilencer对照组相比,P<0.05)。与pcDNA3对照组比较,pcDNA3/CAAP1组SMMC-7721细胞的增殖能力、克隆形成能力、运动能力、迁移和侵袭能力增加,而细胞的凋亡受到抑制(均P<0.05);与pSilencer对照组相比较,shR-CAAP1组SMMC-7721细胞的增殖能力、克隆形成能力、运动能力、迁移和侵袭能力降低,而细胞凋亡增加(P<0.05)。TCGA数据库分析表明CAAP1低表达时肝癌患者的OS优于CAAP1高表达者。结论CAAP1能够促进肝癌细胞SMMC-7721增殖、迁移和侵袭,并抑制其凋亡。