摘要
目的:探讨长链非编码RNA(long noncoding RNA,LncRNA)转移相关肺腺癌转录本1(metastasis-associated lung adenocarcinoma transcript 1,MALAT1)与微小核糖核酸(microRNA,miRNA)-146a相互作用在对子痫前期(preecampsia,PE)滋养细胞功能的影响及机制研究。方法:体外培养绒毛膜癌细胞JEG-3细胞,分4组对JEG-3细胞进行转染,分别为Control、sh-MALATl、miR-146a-5p inhibitor和sh-MALATI+miR-146a-5p inhibitor组,其中sh-MALATl组转染sh-MALATl,miR-146a-5p inhibitor组转染miR-146a-5p inhibitor,sh-MALATl+inhibitor组共同转染sh-MALATl与miR-146a-5p inhibitor,Control组加入等量空载体。通过QPCR检测mRNA水平;生物信息预测MALATl与miR-146a-5p的靶向关系;通过CCK8实验检测各组质粒转染JEG-3细胞后对其增殖能力的影响;利用Western Blot技术检验不同方式处理后,JEG-3细胞内蛋白的表达量变化。结果:sh-MALATl明显降低MALATl并提高miR-146a-5p在JEG-3细胞mRNA水平。sh-MALATl抑制JEG-3细胞增殖能力;miR-146a-5p inhibitor促进细胞增殖能力,减弱sh-MALATl产生的影响。同时,sh-MALATl减弱滋养细胞中TRAF6、VEGR、MMP2蛋白的表达,而miR-146a-5p inhibitor可增强其表达,并减弱sh-MALATl的抑制作用。结论:MALATl与miR-146a可相互作用对子痫前期滋养细胞生物学行为产生影响。
Objective:To investigate the interaction between long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1(MALAT1)and microrna-146a on the function of trophoblast and their mechanisms.Methods:Choriocarcinoma cell line JEG-3 were cultured in vitro and transfected into four groups control,sh-MALAT1,miR-146a-5p inhibitor and sh-MALAT1+miR-146a-5p inhibitor.Among them,sh-MALAT1 group was transfected with sh-MALAT1,miR-146a-5p inhibitor group was transfected with miR-146a-5p inhibitor,sh-MALAT1+inhibitor group was co transfected with sh-MALAT1 and miR-146a-5p inhibitor,control group was added with equal amount of empty vector.The mRNA level was detected by qPCR;the target relationship between MALAT1 and miR-146a-5p was predicted by bioinformation;the proliferation ability of JEG-3 cells was detected by CCK8 experiment after the four groups of plasmids were transfected;Western blot was used to detect the expression of protein in JEG-3 cells after different treatments.Results:sh-MALATl significantly decreased sh-MALATl and increased the mRNA level of miR-146a-5p in the choriocarcinoma cell line JEG-3.sh-MALATl inhibited the proliferation of JEG-3 cells,miR-146a-5p inhibitor promoted the proliferation of JEG-3 cells and weakened the effect of sh-MALATl production.At the same time,sh-MALATl attenuates the expression of TRAF6,VEGR,and MMP2 proteins in trophoblast cells,while miR-146a-5p inhibitor can enhance its expression and reduce the inhibitory effect of sh-MALATl.Conclusion:MALATl and miR-146a can interact to affect the biological behavior of trophoblast cells in preeclampsia.
作者
陈芳荣
龚护民
郑林媚
CHEN Fang-rong;GONG Hu-min;ZHENG Lin-mei(Department of Obstetrics,Hainan General Hospital Hainan Affifliated Hospital of Hainan Medical University,Haikou 570311,China)
出处
《海南医学院学报》
CAS
2020年第12期915-920,共6页
Journal of Hainan Medical University
基金
海南省自然科学基金面上项目(818MS123)
海南省高等学校科学研究项目(Hnky2019-40)。
