Fang Fazhuang, Cheng Ling, Wu Xiaotang, Ye Minfeng, Zhang Huizhong
Department of Hepatobiliary, Pancreatic and Gastric Surgery, Jinhua Guangfu Hospital, Jinhua 321000, People's Republic of China.
Shanghai Engineering Research Center of Pharmaceutical Translation, Shanghai, People's Republic of China.
Cancer Manag Res. 2020 Nov 6;12:11341-11350. doi: 10.2147/CMAR.S256670. eCollection 2020.
Colon cancer (CC) is the third most common cancer with a high rate of incidence and mortality. Therefore, it is highly necessary to explore novel targets of CC.
The miRNA-seq and RNA-seq data of CC were accessed from the TCGA database. Differential analysis was performed using the "edgeR" package to identify differentially expressed miRNAs (DE_miRNAs). The downstream target genes of the target miRNA were then predicted by miRNA target prediction databases to identify the target mRNA. Normal colon cell line CCD-18Co and CC cell lines HCT-116, HT-29, SW620 and SW480 were chosen, and qRT-PCR was conducted to detect expression in these cell lines. qRT-PCR and Western blot were carried out to determine mRNA and protein expression, respectively. Dual-luciferase reporter gene assay was performed to verify the targeting relationship between and 3'UTR. CCK-8 assay and colony formation assay were carried out to detect cell proliferation. Meanwhile, tumor xenograft model in nude mice was constructed to assess CC cell tumorigenic ability in vivo.
was markedly up-regulated in CC tissue. CC cell proliferation and in vivo tumorigenic ability were suppressed by silencing but promoted by over-expression. was significantly down-regulated in cancer tissue. Dual-luciferase reporter gene assay indicated that could bind to 3'UTR. expression was noticeably elevated upon deficiency but significantly inhibited upon over-expression. CCK-8 and colony formation assay suggested that facilitated CC cell proliferation by silencing .
promotes CC cell proliferation by targeted silencing .
结肠癌（CC）是第三大常见癌症，发病率和死亡率都很高。因此，探索结肠癌的新靶点非常必要。
从TCGA数据库获取结肠癌的miRNA测序和RNA测序数据。使用“edgeR”软件包进行差异分析，以鉴定差异表达的miRNA（DE_miRNA）。然后通过miRNA靶标预测数据库预测靶标miRNA的下游靶基因，以鉴定靶标mRNA。选择正常结肠细胞系CCD-18Co和结肠癌细胞系HCT-116、HT-29、SW620和SW480，进行qRT-PCR检测这些细胞系中的表达。分别进行qRT-PCR和蛋白质免疫印迹法以确定mRNA和蛋白质表达。进行双荧光素酶报告基因检测以验证与3'UTR之间的靶向关系。进行CCK-8检测和集落形成检测以检测细胞增殖。同时，构建裸鼠肿瘤异种移植模型以评估结肠癌细胞在体内的致瘤能力。
在结肠癌组织中明显上调。通过沉默抑制结肠癌细胞增殖和体内致瘤能力，但通过过表达促进。在癌组织中明显下调。双荧光素酶报告基因检测表明可以与3'UTR结合。在缺失时表达明显升高，但在过表达时明显受到抑制。CCK-8和集落形成检测表明通过沉默促进结肠癌细胞增殖。
通过靶向沉默促进结肠癌细胞增殖。
