摘要
目的研究microRNA-155(miR-155)对高转移性人食管癌EC109细胞增殖和侵袭等生物学行为的影响。方法以人食管癌EC109基因组DNA为模板,经PCR法扩增miR-155的前体序列,通过BamHⅠ和HindⅢ双酶切后将其亚克隆入真核表达载体pcDNA 3.1(-);然后将构建成功的pcDNA3.1(-)-pri-miR-155载体(命名为p-miR-155)体外瞬时转染人食管癌EC109细胞,利用qRT-PCR(quantitative Real-time PCR)法检测mR-155成熟体的表达水平,并利用MTT实验、克隆形成实验和划痕法检测EC109细胞的增殖、克隆形成以及体外迁移能力。结果成功构建携miR-155的真核表达载体;与空白(Mock)和对照组(p-Ctrl)相比,转染后的EC109细胞过表达miR-155、p-miR-155载体转染组EC109细胞的增殖抑制明显增加(P<0.01),同时克隆形成能力(在100和1 000个细胞/孔接种条件下)明显下降(P<0.01),以及伤口愈合能力下降(P<0.01)。结论过表达miR-155可显著抑制人食管癌EC109细胞的增殖和迁移能力,为食管癌的治疗提供了潜在的策略。
Objective To explore the effect of microRNA-155 （ miR-155 ） on invasion and migration of human esophageal cancer （ Eca） EC109 cells. Methods The pri-miR-155 sequence was amplified from total RNA of Eca EC109 cells by RT-PCR, and cloned into pcDNA3.1 and constructed pcDNA3.1 （-） -pri-miR-155 （ p-miR-155 ） . After p-miR-155 was transfected into EC109 cells, the expression of miR-155 was measured. Cell proliferation, clone formation and migration of EC109 cells were detected by MTT assay, clone formation assay and scratch assay, respectively. Results The expression of miR-155 was apparently increased in EC109 cells after transfection of p-miR-155. Compared with mock-transfected and control cells, cell proliferation, clone formation and cell migration were suppressed in EC109 cells when transfected with p-miR-155 （ P〈0.01） . Conclusion Overexpression of miR-155 is capable of inhibition of EC109 cell proliferation and migration, which may provide a reference for therapy of esophageal cancer.
出处
《广东药科大学学报》
CAS
2017年第1期108-111,126,共5页
Journal of Guangdong Pharmaceutical University
关键词
食管癌
MIR-155
侵袭和转移
治疗
human esophageal cancer
miR-155
invasion and migration
therapy
