miR-122 在结肠癌细胞系HCT116 中的表达 及其对增殖和迁移的影响
| 作者: |
1陈一思,
2易维,
1时昭红
1 武汉市第一医院消化内科，武汉 430022 2 华中科技大学同济医学院附属同济医院综合科，武汉 430030 |
| 通讯: |
时昭红
Email: 292922653@qq.com |
| DOI: | 10.3978/j.issn.2095-6959.2018.10.003 |
摘要
目的：探讨miR-122在结肠癌细胞系HCT116中的表达及对增殖和迁移的影响。方法：qRT-PCR分别检测正常结肠上皮细胞系NCM460和结肠癌细胞系HCT116，HT29，LOVO，SW620中miR-122的表达。将HCT116细胞系分成3组：miR-122过表达组(miR-122组)、阴性对照组(miR-Con组)及空白对照组(Mock组)，经LipofectamineTM 2000分别转染miR-122 mimic，miR-Con序列及空白对照。 CCK-8法及细胞划痕实验分别测定细胞增殖和迁移能力，Western 印迹测定Wnt1和cAMP反应元件结合蛋白(CREB1)表达。结果：miR-122在结肠癌细胞系HCT116，HT29，LOVO及SW620中的表达量低于NCM460。转染后72 h及96 h，miR-122组OD450 nm低于miR-Con组和Mock组(P<0.05)。 miR-122组划痕愈合率低于miR-Con组[(25.4±2.7)% vs (49.3±5.6)%，P<0.05]。miR-122组CREB1蛋白表达量低于miR-Con组(0.34±0.04 vs 1.0±0.05，P<0.01)。miR-122组Wnt1蛋白表达量低于miR-Con组(0.53±0.06 vs 1.0±0.07，P<0.05)。结论：miR-122在结肠癌细胞系HCT116中低表达，过表达miR-122可抑制结肠癌增殖和迁移，其机制可能与CREB1和Wnt1蛋白下调表达有关。
关键词：
miR-122；结肠癌；增殖；迁移
Expression of miR-122 in colon carcinoma cell line HCT116 and its effect on cell proliferation and migration
CorrespondingAuthor: SHI Zhaohong Email: 292922653@qq.com
DOI: 10.3978/j.issn.2095-6959.2018.10.003
Abstract
Objective: To investigate the expression and effect of miR-122 on cell proliferation and migration in colon carcinoma cell line HCT116. Methods: The qRT-PCR was used to detect the expression of miR-122 in colon epithelial cell line NCM460 and four colon carcinoma cell lines, HCT116, HT29, LOVO and SW620. The HCT116 cell line was divided into three groups, miR-122 over-expression group (miR-122 group), negative control group (miR-Con group) and blank control group (Mock group), which was transfect with miR-122 mimic, miR-Con sequence and PBST by LipofectamineTM 2000. The proliferation and migration ability was analyzed using CCK-8 and wound scratch assay. The expression of Wnt1 and CREB1 protein was analyzed by Western blot. Results: The expression of miR-122 in four colon carcinoma cell lines HCT116, HT29, LOVO, SW620, was significantly lower than that in colon epithelial cell line NCM460. The OD450 nm value of miR-122 group was significantly lower than that in miR-Con group and Mock group after transfect for 72 h and 96 h, respectively (P<0.05). Scar healing rate of miR-122 group was significantly lower than that in miR-Con group [(25.4±2.7)% vs (49.3±5.6)%, P<0.05]. The expression level of CREB1 protein of miR-122 group was significantly lower than miR-Con group (0.34±0.04 vs 1.0±0.05, P<0.01). The expression level of Wnt1 protein of miR-122 group was significantly lower than miR-Con group (0.53±0.06 vs 1.0±0.07, P<0.05). Conclusion: miR-122 was down-regulated expressed in colon carcinoma cell lines. Over-expression of miR-122 could inhibit cell proliferation and migration, which may be associated with down-regulation of Wnt1 and CREB1 protein.
Keywords：
miR-122; colon carcinoma; proliferation; migration