目的:研究miR495、miR551a干扰质粒对SGC7901胃癌细胞中促肝细胞再生磷酸酶-3(phosphatase of regenerating liver-3,PRL-3)表达的影响,探讨基因干扰在胃癌治疗中的价值.方法:分别体外培养稳定转染后的SGC7901细胞系及未经处理的SGC790...目的:研究miR495、miR551a干扰质粒对SGC7901胃癌细胞中促肝细胞再生磷酸酶-3(phosphatase of regenerating liver-3,PRL-3)表达的影响,探讨基因干扰在胃癌治疗中的价值.方法:分别体外培养稳定转染后的SGC7901细胞系及未经处理的SGC7901细胞系,取对数生长期瘤细胞,0.5mL(1×107/mL)细胞悬液接种于Balb/ca(nu/nu)裸鼠腹腔内,在SPF条件下饲养1mo后处死,观察裸鼠成瘤率、瘤体生长情况及腹膜转移等情况,并行组织病理学检查.取各组部分移植瘤行荧光定量PCR检测,对比各组miR495、miR551a及PRL-3mRNA的相对表达水平.结果:各组裸鼠成瘤率均100%,其中两实验组裸鼠一般情况及生存期均好于对照组.取各组移植瘤行荧光定量PCR检测显示,两实验组裸鼠miRNA表达量显著高于对照组,PRL-3mRNA表达量低于对照组.转染质粒组胃癌迁移能力明显减弱.结论:转染靶向干扰PRL-3表达的miR495、mi551a真核质粒可以明显抑制胃癌细胞体内转移侵袭能力.展开更多
为给新矮秆种质系山农495的利用提供依据,利用田间观察和室内赤霉酸鉴定与分子标记分析相结合的方法,进行矮秆性状的遗传分析和矮秆基因分子标记定位。结果表明,山农495的矮秆性状受位于4BS染色体上的1对隐性主效基因控制,且为赤霉酸不...为给新矮秆种质系山农495的利用提供依据,利用田间观察和室内赤霉酸鉴定与分子标记分析相结合的方法,进行矮秆性状的遗传分析和矮秆基因分子标记定位。结果表明,山农495的矮秆性状受位于4BS染色体上的1对隐性主效基因控制,且为赤霉酸不敏感型。在供试的1 589个SSR引物中,有2个基因组SSR引物(Gwm113196、Wmc 511191)和1个EST-SSR引物(Dupw23210)在优选小群体中能扩增出多态性谱带。利用BC1回交群体和F2群体进行连锁标记分析,Gwm113196、Wmc 511191和Dupw23210与山农495矮秆基因的连锁距离分别为3.9、5.2、21.6 c M和4.1、5.0、19.8 c M。展开更多
MicroRNAs （miRNAs） are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast canc...MicroRNAs （miRNAs） are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast cancer. In the present study, we found that miR-495 was markedly up-regulated in clinical breast cancer samples by quantitative real time-PCR （qRT-PCR）. Junctional adhesion molecule A （JAM-A） was predicted to be a potential target of miR-495 by bioinformatics analysis and was subsequently verified by luciferase assay and Western blotting. JAM-A was found to be negatively correlated with the migration of breast cancer cells through loss-of-function and gain-offunction assays, and the inhibition of JAM-A by miR- 495 promoted the migration of MCF-7 and MDA-MB-231 cells. Furthermore, overexpression of JAM-A could restore miR-495-induced breast cancer cell migration. Taken together, our findings suggest that miR-4g5 could facilitate breast cancer progression through the repression of JAM-A, making this miRNA a potential therapeutic target.展开更多
文摘目的:研究miR495、miR551a干扰质粒对SGC7901胃癌细胞中促肝细胞再生磷酸酶-3(phosphatase of regenerating liver-3,PRL-3)表达的影响,探讨基因干扰在胃癌治疗中的价值.方法:分别体外培养稳定转染后的SGC7901细胞系及未经处理的SGC7901细胞系,取对数生长期瘤细胞,0.5mL(1×107/mL)细胞悬液接种于Balb/ca(nu/nu)裸鼠腹腔内,在SPF条件下饲养1mo后处死,观察裸鼠成瘤率、瘤体生长情况及腹膜转移等情况,并行组织病理学检查.取各组部分移植瘤行荧光定量PCR检测,对比各组miR495、miR551a及PRL-3mRNA的相对表达水平.结果:各组裸鼠成瘤率均100%,其中两实验组裸鼠一般情况及生存期均好于对照组.取各组移植瘤行荧光定量PCR检测显示,两实验组裸鼠miRNA表达量显著高于对照组,PRL-3mRNA表达量低于对照组.转染质粒组胃癌迁移能力明显减弱.结论:转染靶向干扰PRL-3表达的miR495、mi551a真核质粒可以明显抑制胃癌细胞体内转移侵袭能力.
文摘为给新矮秆种质系山农495的利用提供依据,利用田间观察和室内赤霉酸鉴定与分子标记分析相结合的方法,进行矮秆性状的遗传分析和矮秆基因分子标记定位。结果表明,山农495的矮秆性状受位于4BS染色体上的1对隐性主效基因控制,且为赤霉酸不敏感型。在供试的1 589个SSR引物中,有2个基因组SSR引物(Gwm113196、Wmc 511191)和1个EST-SSR引物(Dupw23210)在优选小群体中能扩增出多态性谱带。利用BC1回交群体和F2群体进行连锁标记分析,Gwm113196、Wmc 511191和Dupw23210与山农495矮秆基因的连锁距离分别为3.9、5.2、21.6 c M和4.1、5.0、19.8 c M。
文摘目的 探讨miR-495通过靶向 Notch1信号调控高糖诱导的视神经节细胞(RGCs)凋亡的效果与机制。方法 将高糖培养的RGC-5细胞系随机分为三组:空白组、miR-495阴性对照(miR NC)和miR-495组。miR-NC组与miR-495组分别转染miR-495模拟物阴性对照(miR NC)与miR-495 模拟物(mimic),空白组不进行转染。采用qRT-PCR检测miR-495表达,CCK-8法检测细胞增殖指数,流式细胞术检测检测细胞周期,流式细胞术检测细胞凋亡指数,Western Blot检测Notch1蛋白表达。结果 转染后24 h与36 h,细胞增殖指数显著高于空白组与miR-NC组( P <0.05);细胞凋亡指数显著低于空白组与miR-NC组( P <0.05);转染后36 h,miR-495组的G1期比率相对于miR-NC组和空白组显著降低( P <0.05),S期+G2期比率则显著增加( P <0.05)。转染后24 h与36 h,miR-495组的Notch1蛋白相对表达量显著低于miR-NC组和空白组,空白组与miR-NC组比较差异无统计学意义( P >0.05)。结论 miR-495能减轻高糖诱导的RGCs损伤,促进RGCs增殖,抑制Notch1信号通路,从而调节细胞周期,抑制RGCs凋亡。
文摘MicroRNAs （miRNAs） are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast cancer. In the present study, we found that miR-495 was markedly up-regulated in clinical breast cancer samples by quantitative real time-PCR （qRT-PCR）. Junctional adhesion molecule A （JAM-A） was predicted to be a potential target of miR-495 by bioinformatics analysis and was subsequently verified by luciferase assay and Western blotting. JAM-A was found to be negatively correlated with the migration of breast cancer cells through loss-of-function and gain-offunction assays, and the inhibition of JAM-A by miR- 495 promoted the migration of MCF-7 and MDA-MB-231 cells. Furthermore, overexpression of JAM-A could restore miR-495-induced breast cancer cell migration. Taken together, our findings suggest that miR-4g5 could facilitate breast cancer progression through the repression of JAM-A, making this miRNA a potential therapeutic target.
