miR-143调控基质金属蛋白酶-13表达对骨肉瘤细胞迁移及侵袭的影响
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作者Author单位AddressE-Mail
李斌 LI Bin 张家口市第一医院骨一科, 河北 张家口 075000 The First Department of Orthopaedics, Zhangjiakou First Hospital, Zhangjiakou 075000, Hebei, China 15132315839@163.com 
李战鹏 LI Zhan-peng 张家口市第一医院骨一科, 河北 张家口 075000 The First Department of Orthopaedics, Zhangjiakou First Hospital, Zhangjiakou 075000, Hebei, China  
连振刚 LIAN Zhen-gang 张家口市第一医院骨一科, 河北 张家口 075000 The First Department of Orthopaedics, Zhangjiakou First Hospital, Zhangjiakou 075000, Hebei, China  
期刊信息:《中国骨伤》2023年,第36卷,第11期,第1075-1080页
DOI:10.12200/j.issn.1003-0034.2023.11.013
基金项目:张家口市科技计划(编号:1711042H)
中文摘要:

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50 μg 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50 μg miR-143 mimic及10 μg 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。
【关键词】骨肉瘤  miR-143  基质金属蛋白酶-13  迁移  侵袭
 
Effects of miR-143 on the migration and invasion of osteosarcoma cells by regulating MMP-13 expression
ABSTRACT  

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 50 μg 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 50 μg miR-143 mimic and 10 μg 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 (1 000.01±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.
KEY WORDS  Osteosarcoma  miR-143(MicroRNA)  Matrix metalloproteinase(MMP)-13  Migration  Invasion
 
引用本文,请按以下格式著录参考文献:
中文格式:李斌,李战鹏,连振刚.miR-143调控基质金属蛋白酶-13表达对骨肉瘤细胞迁移及侵袭的影响[J].中国骨伤,2023,36(11):1075~1080
英文格式:LI Bin,LI Zhan-peng,LIAN Zhen-gang.Effects of miR-143 on the migration and invasion of osteosarcoma cells by regulating MMP-13 expression[J].zhongguo gu shang / China J Orthop Trauma ,2023,36(11):1075~1080
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