CAJ | 학술논문

背景：抑癌基因表达抑制在肿瘤发生、发展过程中起重要作用。一些microRNAs可通过调节抑癌基因的表达影响肿瘤发生。目的：探讨miR-483-3p对结直肠癌肝癌缺失基因1（DLC1）表达的靶向调节作用。方法：纳入2012年10月~2013年4月南京鼓楼医院收治的结直肠癌患者16例，采用蛋白质印迹法检测癌组织及其相应癌旁非癌组织的DLC1表达，qRT-PCR检测miR-483-3p表达。构建含DLC13’非翻译区（3’UTR）的双荧光素酶报告基因质粒，在人结肠癌细胞株HCT116中验证miR-483-3p对DLC1表达的调节作用。以miR-483-3p mimic转染HEK293T细胞，采用蛋白质印迹法检测DLC1表达；以miR-483-3p mimic转染HCT116细胞，采用CCK-8实验检测细胞增殖。结果：结直肠癌组织的DLC1表达水平显著低于癌旁非癌组织，miR-483-3p表达水平显著高于癌旁非癌组织（ P<0.05）。miR-483-3p mimic可靶向结合DLC1的3’UTR而抑制其表达。转染miR-483-3p mimic的HCT116细胞增殖能力显著增强（ P<0.05）。结论：DLC1是miR-483-3p的靶基因，miR-483-3p可在转录后水平抑制DLC1表达，参与促进结直肠癌发生。
배경：억암기인표체억제재종류발생、발전과정중기중요작용。일사microRNAs가통과조절억암기인적표체영향종류발생。목적：탐토miR-483-3p대결직장암간암결실기인1（DLC1）표체적파향조절작용。방법：납입2012년10월~2013년4월남경고루의원수치적결직장암환자16례，채용단백질인적법검측암조직급기상응암방비암조직적DLC1표체，qRT-PCR검측miR-483-3p표체。구건함DLC13’비번역구（3’UTR）적쌍형광소매보고기인질립，재인결장암세포주HCT116중험증miR-483-3p대DLC1표체적조절작용。이miR-483-3p mimic전염HEK293T세포，채용단백질인적법검측DLC1표체；이miR-483-3p mimic전염HCT116세포，채용CCK-8실험검측세포증식。결과：결직장암조직적DLC1표체수평현저저우암방비암조직，miR-483-3p표체수평현저고우암방비암조직（ P<0.05）。miR-483-3p mimic가파향결합DLC1적3’UTR이억제기표체。전염miR-483-3p mimic적HCT116세포증식능력현저증강（ P<0.05）。결론：DLC1시miR-483-3p적파기인，miR-483-3p가재전록후수평억제DLC1표체，삼여촉진결직장암발생。
Background:Suppression of tumor suppressor genes plays a key role in the pathogenesis and progress of tumors. Some microRNAs may contribute to tumorigenesis by regulating tumor suppressor genes. Aims:To investigate the targeted regulatory effect of miR-483-3p on deleted in liver cancer 1(DLC1)gene in colorectal cancer. Methods:Sixteen patients with colorectal cancer admitted from October 2012 to April 2013 at Nanjing Drum Tower Hospital were enrolled. Expression of DLC1 in cancerous and adjacent noncancerous tissues was determined by Western blotting,and expression of miR-483-3p was determined by qRT-PCR. Dual luciferase reporter gene plasmid containing the 3’untranslated region(3’UTR)of DLC1 was constructed to validate the regulation of DLC1 by miR-483-3p in human colon cancer cell line HCT116. MiR-483-3p mimic was transfected into HEK293T cells and expression of DLC1 was determined by Western blotting;MiR-483-3p mimic was transfected into HCT116 cells and cell proliferation was measured by CCK-8 assay. Results:Expression of DLC1 was significantly lower in cancerous tissue than in noncancerous tissue,while expression of miR-483-3p was significantly higher in cancerous tissue than in noncancerous tissue(P<0. 05). MiR-483-3p mimic reduced the expression of DLC1 through directly binding to the 3’UTR of DLC1. Transfection of miR-483-3p mimic enhanced the proliferation of HCT116 cells significantly(P<0. 05). Conclusions:DLC1 is a target gene of miR-483-3p. MiR-483-3p might promote the development of colorectal cancer by down-regulating DLC1 expression at post-transcriptional level.