miR-143过表达对骨肉瘤143B细胞侵袭和迁移能力的影响  

Effect of miR-143 overexpression on the invasion and migration of osteosarcoma 143B cells

作　　者：李勇[1] 齐巍[2] 王保苍[1] 范超[3] 

机构地区：[1]唐山市第二医院骨病科,河北唐山063000 [2]唐山市第二医院创伤科,河北唐山063000 [3]唐山市第二医院理疗科,河北唐山063000

出　　处：《广东医学》2017年第12期1809-1813,共5页Guangdong Medical Journal

基　　金：河北省2016年度医学科学研究重点课题(编号:20160239)

摘　　要：目的研究miR-143过表达对人成骨肉瘤143B细胞侵袭及迁移能力的影响。方法通过慢病毒携带miR-143过表达载体转染骨肉瘤143B细胞,实验分为3组。实验组转染miR-143过表达序列,对照组转染阴性对照NC序列,空白组为空白PBS。分别采用实时荧光定量PCR、Western blot、Transwell小室侵袭实验和细胞划痕实验检测各组基因、蛋白的表达以及细胞侵袭和迁移能力。结果实验组miR-143基因和蛋白的表达,相较于对照组和空白组显著增加(P<0.05),而基质金属蛋白酶-13(MMP-13)基因和蛋白的表达相较于对照组和空白组显著减少(P<0.05);实验组穿膜细胞数显著低于对照组和空白组(P<0.05);实验组细胞划痕愈合率显著低于对照组和空白组(P<0.05)。结论 miR-143过表达通过靶向MMP-13基因,进一步抑制成骨肉瘤143B细胞侵袭及迁移能力。Objective To study the effect of miR - 143 overexpression on the invasion and migration of human osteosareoma 143B cells. Methods The miR - 143 overexpressing vector was carried by lentivirus and transfected into osteosarcoma 143B cells. The experiment was divided into, experimental group （transfected with miRNA - 143 overexpression sequence）, control group （transfeeted with negative control -NC sequence） , and blank group （PBS）. Real - time PCR was used to detect the expression of mRNA. The protein expression was detected by Western blot. Transwell invasion assay was used to detect the invasive ability of the cells. Cell migration was measured by cell scratch test. Results The expression of miR - 143 mRNA and protein in the experimental group was significantly higher than control group and blank group （ P 〈 0. 05 ）. However, the mRNA and protein expression of MMP - 13 was significantly down - regulated in experimental group than control group and blank group （ P 〈 0. 05 ）. The number of transmembrane ceils and healing rate of scratches in the experimental group were significantly lower than control group and blank group （ P 〈 0. 05 ）. Conclusion MiR - 143 overexpression inhibits invasion and migration of osteoblast 143B cells by acting on MMP - 13 target genes.

关 键 词：骨肉瘤 微小RNA 基质金属蛋白酶-13 细胞侵袭 细胞迁移 

分 类 号：R738.1[医药卫生—肿瘤]