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作 者:卢敏[1] 郑相涛 毛华[1] 沙卫红[3] LU Min;ZHENG Xiangtao;MAO Hua;SHA Weihong(De⁃partment of Gastroenterology,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China)
机构地区:[1]南方医科大学珠江医院消化内科,广州510282 [2]南方医科大学第二临床医学院,广州510282 [3]广东省人民医院消化内科,广东省医学科学院,广州510080
出 处:《实用医学杂志》2020年第10期1292-1298,共7页The Journal of Practical Medicine
基 金:广东省自然科学基金资助项目(编号:2014A030313543)。
摘 要:目的利用生物信息学对结肠癌差异表达基因进行分析,探索结肠癌治疗的新靶点,并通过RNA干扰了解其对结肠癌细胞增殖及侵袭迁移机制研究。方法从GEO数据库下载结肠癌基因芯片数据库,利用GEO2R筛选差异表达基因,并通过RT⁃qPCR和Western Blot检测RNA干扰对靶基因表达变化。CCK⁃8测定LOVO细胞的增殖和活力,Transwell小室检测细胞侵袭和迁移力,流式细胞术检测细胞周期变化。结果GEO2R筛选差异表达基因BMI1,RNA干扰靶基因后细胞增殖及侵袭迁移能力下降,细胞周期停滞于G1期,S期明显缩短,其下游P16ink4a基因表达增高,E-钙粘蛋白(E⁃cadherin)的表达增加,而波形蛋白(Vimentin)的表达减少。结论利用生物信息学分析可找出结肠癌差异表达靶基因BMI1,RNA干扰BMI1后可能通过P16ink4a表达上调和EMT下调抑制结肠癌细胞的增殖、侵袭和迁移,为结肠癌的基因治疗提供新的思路。Objective To explore new targets of colon cancer treatment of differential genes in colon can⁃cer by bioinformatics analysis and to understand the mechanism of proliferation,invasion and migration of colon cancer cells by RNA interference.Methods The gene chip database of colon cancer was downloaded form the GEO database and differential genes were screened by GEO2R.RT⁃qPCR and western blot were used to detect the target gene expression caused by RNA interference.The proliferation and viability of colon cancer cells was determined by Cell Counting Kit⁃8 assay.Cell migration was determined by transwell assay.Cell cycle progression was analyzed by flow cytometry.Results BMI1,a differential gene,was screened by GEO2R.The proliferation and migration declined after BMI1 knockdown,and G1 phase arrested and S phase shortened in cell cycle.Also,knockdown of BMI1 increased the expressionof its downstream effector P16ink4a,and increased the E⁃cadherin expression but suppressed the Vimentin expression.Conclusions Bioinformatics can reveal the differential gene BMI1 in colon cancer.RNA interference of BMI1 may inhibit the proliferation,invasion and migration of colon cancer cells through the upregulation of p16INK4a and inhibition of EMT,which provide a new potential target of colon cancer.
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