摘要
目的:探究miR-143对基质金属蛋白酶-13(matrix metalloproteinase,MMP-13)表达的调节作用及其对骨肉瘤细胞迁移及侵袭能力的影响。方法:采用96孔板培养小鼠骨肉瘤细胞系143B细胞,设立空白组、阴性组、阳性组、干预组。空白组不做特殊处理,阳性组加入50μg miR-143 mimic,阴性组加入等量mimic NC(miR-143 mimic的对照序列),干预组加入50μg miR-143 mimic及10μg MMP-13蛋白,继续培养3~6 h,最后吸出血清处理0.5 h。分别采用荧光定量PCR和Western-blot测定各组miR-143和MMP-13蛋白表达,采用Transwell、划痕实验测定细胞的侵袭与迁移能力。结果:阳性组及干预组MMP-13蛋白表达量明显低于空白组,阳性组低于干预组(P<0.05);空白组、阴性组、阳性组及干预组每个视野的侵袭细胞数均值分别为(1 000.01±44.77)、(959.25±46.32)、(245.04±4.33)、(634.06±33.78)个;阳性组及干预组划痕愈合率均明显低于空白组,阳性组低于干预组(P<0.05)。结论:MMP-13是miR-143作用的一个靶点,可以通过抑制MMP-13表达降低骨肉瘤细胞的迁移及侵袭能力。
Objective To explore the effect of miR-143 regulating matrix metalloproteinase(MMP)-13 expression on migration and invasion of osteosarcoma cells.Methods The mouse osteosarcoma cell line 143B cells were cultured in 96-well plates,and blank group,negative group,positive group,and intervention group were set up.Then,the blank group did no treatment 50μg miR-143 mimic was added to positive group,negative group added equal mimic NC(control sequence of miR-143 mimic),the intervention group was added 50μg miR-143 mimic and 10μg MMP-13 protein,all groups continued to culture for 3 to 6 hours,and finally the serum was aspirated to treat for half an hour.The protein expressions of miR-143 and MMP-13 in each group were measured by fluorescence quantitative PCR experiment and Western blot experiment,respectively,and the invasion and migration abilities of cells were measured by Transwell and scratch experiments.Results The expression of MMP-13 protein in the positive group and the intervention group was significantly lower than that in the blank group,and the positive group was lower than the intervention group(P<0.05);The mean numbers of invasive cells in blank group,negative group,positive group and intervention group were(1000.01±44.77),(959.25±46.32),(245.04±4.33),(634.06±33.78)cells/field,respectively;the scratch healing rate of the positive group and the intervention group was significantly lower than that of the blank group,and the positive group was lower than the intervention group(P<0.05).Conclusion MMP-13 is a target of miR-143,which can reduce the migration and invasion ability of osteosarcoma cells by inhibiting the expression of MMP-13.
作者
李斌
李战鹏
连振刚
LI Bin;LI Zhan-peng;LIAN Zhen-gang(The First Department of Orthopaedics,Zhangjiakou First Hospital,Zhangjiakou 075000,Hebei,China)
出处
《中国骨伤》
CAS
CSCD
2023年第11期1075-1080,共6页
China Journal of Orthopaedics and Traumatology
基金
张家口市科技计划(编号:1711042H)。
