目的探讨基因枪转导突变特异性K-ras si RNA对胰腺癌细胞生长的抑制作用。方法根据胰腺癌细胞系突变特征,设计突变特异性K-ras si RNA;应用基因枪将si RNA转导入胰腺癌细胞系,提取总RNA和胞浆蛋白;应用逆转录-聚合酶链反应(RT-PCR)和免...目的探讨基因枪转导突变特异性K-ras si RNA对胰腺癌细胞生长的抑制作用。方法根据胰腺癌细胞系突变特征,设计突变特异性K-ras si RNA;应用基因枪将si RNA转导入胰腺癌细胞系,提取总RNA和胞浆蛋白;应用逆转录-聚合酶链反应(RT-PCR)和免疫印迹法(Westernblot)方法,检测基因枪转导突变特异性K-ras si RNA对突变型K-ras基因表达的干扰作用;行细胞爬片K-ras p21蛋白免疫组织化学染色,探讨si RNA对K-ras p21蛋白表达的抑制作用;采用CCK-8(cell counting kit-8)活细胞计数法,绘制转基因前后细胞生长曲线,观察si RNA的抑瘤效果。结果RT-PCR结果显示,K-ras突变特异性si RNA,能有效抑制K-ras基因表达(P<0.05);Westernblot和免疫组织化学结果表明,K-ras p21蛋白的表达明显降低(P<0.05);细胞生长曲线显示,转导si RNA组较对照组细胞生长明显受抑制(P<0.05)。结论基因枪可以有效地对小片段双链si RNA进行细胞内转导;突变特异性K-ras si RNA,可显著下调胰腺癌细胞系突变型K-ras mRNA及K-ras p21蛋白表达,并能有效抑制肿瘤细胞的增殖。展开更多
[Objective] The experiment aimed to explore physiological and biochemical changes of leaves after plants were mutated. [Method] A rice double mutant with stripes on stems, leaves and spikelets were taken as experiment...[Objective] The experiment aimed to explore physiological and biochemical changes of leaves after plants were mutated. [Method] A rice double mutant with stripes on stems, leaves and spikelets were taken as experimental materials to study the enzyme activity changes in different growth stages and amino acid variation in rice. [ Result] The SOD activity in mutant was higher than that in wild plant at tillering metaphase, but lower than that in wild type before heading stage and late flowering; the POD activity in three stages increased firstly then declined and the activity showed highest maximal activity at before heading stage. However, the POD activity in wild type showed the opposite change trend; the CAT activity presented degression at three stages, especially high at tillering metaphase, but reverse changes in wild type; the MDA activity decreased at three stages, but it was still higher than that in wild type, besides, the soluble sugar content of mutant was lower, but total amino acid content was increased. [ Conclusion] The expression of mutant characteristics was correlated with SOD, POD, CAT and MDA activity Changes and these changes made the mutant survive and rice quality change at last.展开更多
基金Supported by Program for New Century Excellent Talents in University(NCET-04-0907)the Innovative Research Team in University (IRT0453)~~
文摘[Objective] The experiment aimed to explore physiological and biochemical changes of leaves after plants were mutated. [Method] A rice double mutant with stripes on stems, leaves and spikelets were taken as experimental materials to study the enzyme activity changes in different growth stages and amino acid variation in rice. [ Result] The SOD activity in mutant was higher than that in wild plant at tillering metaphase, but lower than that in wild type before heading stage and late flowering; the POD activity in three stages increased firstly then declined and the activity showed highest maximal activity at before heading stage. However, the POD activity in wild type showed the opposite change trend; the CAT activity presented degression at three stages, especially high at tillering metaphase, but reverse changes in wild type; the MDA activity decreased at three stages, but it was still higher than that in wild type, besides, the soluble sugar content of mutant was lower, but total amino acid content was increased. [ Conclusion] The expression of mutant characteristics was correlated with SOD, POD, CAT and MDA activity Changes and these changes made the mutant survive and rice quality change at last.
