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作 者:冯吉[1] 刘琴[1] 刘磊[1] 赖姝婕[1] 陈东风[1]
机构地区:[1]第三军医大学大坪医院野战外科研究所消化内科,重庆400042
出 处:《第三军医大学学报》2016年第10期1160-1165,共6页Journal of Third Military Medical University
摘 要:目的研究口腔癌过表达蛋白1(oral cancer overexpressed protein 1,ORAOV1)在人结肠癌组织中的表达变化并探讨其对结肠癌细胞增殖能力、细胞周期及早期凋亡的影响。方法收集人结肠癌及癌旁组织标本,分别以qRT-PCR及Western blot检测ORAOV1在mRNA及蛋白水平的表达变化情况;采用RNAi技术获得ORAOV1低表达的Lo Vo及SW480细胞株,并使用Western blot对干扰结果进行鉴定;采用CCK-8检测干扰ORAOV1表达后两种结肠癌细胞增殖能力的变化;并采用流式细胞术检测干扰ORAOV1表达对结肠癌细胞周期及早期凋亡的影响。结果 qRT-PCR及Western blot结果分别显示结肠癌组织中的ORAOV1 mRNA及蛋白水平高于癌旁组织(P<0.05);Western blot检测证实Lo Vo及SW480细胞中ORAOV1干扰成功;在这两种结肠癌细胞株中,CCK-8检测结果显示,干扰ORAOV1可抑制细胞增殖能力;流式细胞术结果显示,干扰ORAOV1可使处于S期比例增加(P<0.05)、早期凋亡比例增加(P<0.05)。结论结肠癌组织中的ORAOV1表达增加,干扰ORAOV1可降低结肠癌细胞的增殖能力,可能与S期阻滞及细胞早期凋亡增加有关。Objective To determine the expression of oral cancer overexpressed protein 1( ORAOV1) in colon cancer tissue and investigate its effect of ORAOV1 on the proliferation,cell cycle and apoptosis in colon cancer cell lines. Methods The mRNA and protein levels of OVAOV1 in 6 tissue samples of human colon cancer and corresponding para-cancer tissues were detected by qRT-PCR and Western blotting. The expression of ORAOV1 in Lo Vo cells and SW480 cells was down-regulated by RNAi technique,and the interference results were confirmed by Western blotting. The proliferation of 2 colon cancer cell lines with different ORAOV1 expression levels was detected by CCK-8 assay. Cell cycle and apoptosis were analyzed by flow cytometry. Results The expression of ORAOV1 at mRNA and protein levels was significantly higher in the colon cancer tissues than the para-cancer tissues( P〈 0. 05). Western blotting indicated that the interference to ORAOV1 was successful in Lo Vo cells and SW480 cells. Compared with the cells with normal ORAOV1 expression,the proliferation of the cells with low ORAOV1 expression was inhibited.ORAOV1 interference increased the percentage of cells at S stage( P〈 0. 05) and early apoptotic ratio( P〈 0. 05). Conclusion Expression of ORAOV1 is increased in colon cancer,and the proliferation of colon cancer cells with ORAOV1 knockdown is inhibited through increasing the S stage arrest and early apoptosis.
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