miR-363靶向作用于CTHRC1对结直肠癌细胞增殖和迁移的影响

Influence of miR-363 on proliferation and migration of colorectal cancer cell by targeting CTHRC1

目的观察微小RNA(microRNA,miRNA)-363在结直肠癌组织中的表达及其对结肠癌细胞增殖和迁移的影响,并探讨其可能的机制。方法选择96例结直肠癌患者手术切除后癌组织、癌旁正常组织标本及6种结肠癌细胞株与正常结直肠上皮细胞株。实时荧光定量PCR(qRT-PCR)检测miR-363在结直肠癌组织和各种细胞株中的表达。以miR-363模拟物(mimics)、miR-363抑制剂(inhibitor)及空白对照分别转染结直肠癌细胞HCT116,采用细胞计数试剂盒(CCK8)法和细胞划痕实验分别检测转染miR-363 mimics和miR-363 inhibitor后结直肠癌细胞HCT116的增殖能力和SW480、SW620迁移能力,Western blot法检测结直肠癌细胞HCT116中胶原三股螺旋重复蛋白1(CTHRC1)的表达。结果与癌旁组织比较,miR-363在结直肠癌组织表达显著下调;与正常结直肠上皮细胞株比较,6种结直肠癌细胞株中miR-363表达显著下调,差异均有统计学意义(P均<0.05)。转染miR-363 mimics可显著抑制结直肠癌细胞HCT116的增殖和SW480、SW620迁移能力。与转染空白对照比较,转染miR-363 mimics后结直肠癌细胞HCT116中CTHRC1的相对表达量显著减低,而转染miR-363 inhibitor后结直肠癌细胞HCT116中的CTHRC1相对表达量显著增加,差异均有统计学意义(P均<0.05),表明miR-363可抑制CTHRC1蛋白的表达。结论 miR-363可抑制结直肠癌细胞的增殖和迁移能力,其机制可能与靶向作用于CTHRC1蛋白有关。

Objective To observe the expression of microRNA（ miRNA）-363 in colorectal cancer tissues and the influence on proliferation and migration of colorectal cancer cells and explore its potential mechanism. Methods The specimens of cancerous tissues and paracancerous tissues resected by operation in 96 cases of colorectal cancer and 6 colon cancer cell lines and normal colorectal epithelial cell strain were selected. Real-time quantitative PCR（ qRT-PCR） method was used to detect the expressions of miRNA-363 in colorectal cancer tissues and different cell lines. After transfection of colorectal cancer cells with miR-363 mimics,miR-363 inhibitor and blank control,cell counting kit-8（ CCK8） essay and cell wound scratch assay were used to respectively detect the proliferation ability of rectal cancer HCT116 cells and migration abilities of rectal cancer SW480 and SW620 cells. Western blot method was used to detect the expression of collagen three stranded helix repeat containing 1（ CTHRC1） in colorectal cancer HCT116 cells. Results Compared with paracancerous tissues,the expression of miR-363 in colorectal cancer tissues was down-regulated significantly（ P〈0. 05）.Comparison with normal colorectal epithelial cell strain,the expressions of miR-363 in 6 colon cancer cell lines were downregulated significantly（ all P〈0. 05）. After being transfeced with miR-363 mimics,the proliferation ability of HCT116 cells and migration abilities of SW480 and SW620 cells can be significantly inhibited. Compared with transfecting blank control,the relative expression level of CTHRC1 in colorectal cancer HCT116 cells after transfection of miR-363 mimics significantly decreased,while it significantly increased after transfection of miR-363 inhibitor. These results show that miR-363 can inhibit the expression of CTHRC1. Conclusion miR-363 could inhibit the proliferation and migration abilities of colorectal cancer cells,and the mechanism may be related to its targeting effect on CTHRC1 protein