摘要
目的研究长链非编码RNA运动神经元和胰腺同源盒1反义RNA1(lncRNA MNX1-AS1)在结直肠癌组织中的表达及其临床意义,并探讨MNX1-AS1在结直肠癌中发挥的生物学功能。方法采用实时荧光定量PCR(qRT-PCR)检测MNX1-AS1在60例结直肠癌组织和癌旁组织中的表达水平,并分析结直肠癌组织中MNX1-AS1的表达水平与患者临床病理参数的关系。MNX1-AS1 siRNA转染结直肠癌细胞系SW480抑制MNX1-AS1的表达后,采用CCK8实验检测细胞增殖能力,流式细胞仪检测细胞周期变化,Transwell实验检测细胞的转移能力,Western blotting检测细胞中细胞周期蛋白D1(cyclin D1)、细胞周期素依赖性蛋白激酶4(CDK4)、基质金属蛋白酶-7(MMP7)蛋白的表达。结果qRT-PCR结果显示MNX1-AS1在结直肠癌组织中的表达高于癌旁组织(P<0.05),MNX1-AS1的表达水平与TNM分期、淋巴结转移和分化程度有关(P<0.05)。干扰SW480细胞中MNX1-AS1的表达后,细胞增殖和转移能力均下降(P<0.05),细胞周期阻滞在G0/G1期(P<0.05),细胞中cyclin D1、CDK4和MMP7蛋白表达减少(P<0.05)。结论MNX1-AS1在结直肠癌组织中呈高表达,且其表达水平与TNM分期、淋巴结转移和分化程度有关。调控cyclin D1、CDK4、MMP7蛋白的表达可能是MNX1-AS1影响结直肠癌细胞增殖和转移的机制之一。
Objective To investigate the expression and clinical significance of long non-coding RNA:Motor Neuron and Pancreas Homeobox 1 antisense RNA1(lncRNA MNX1-AS1),in colorectal cancer tissues,and to explore its biological function in colorectal cancer.Methods Real-time fluorescent quantitative(PCRqRT-PCR)was used to detect the expression level of MNX1-AS1 in 60 cases of colorectal cancer tissues and adjacent tissues.The relationship between the expression level of MNX1-AS1 and the clinicopathological parameters of colorectal cancer tissues was analyzed.After MNX1-AS1 siRNA was transfected into a colorectal cancer cell line SW480 to inhibit the expression of MNX1-AS1,multiple analyses were conducted:CCK8 assay was used to detect cell proliferation;flow cytometry was used to detect cell cycle;transwell assay was used to detect cell transfer ability;western blotting was used to detect the expression of cyclinD1,cyclin-dependent kinases 4(CDK4)and matrix metalloproteinase 7(MMP7)proteins.Results The results of qRT PCR showed that the expression of MNX1-AS1 in colorectal cancer tissues was higher than that in adjacent tissues(P<0.05).The expression level of MNX1-AS1 was associated with TNM staging,lymph node metastasis and degree of differentiation(P<0.05).After interfering with the expression of MNX1-AS1 in SW480 cells,cell proliferation and metastasis were decreased(P<0.05),cell cycle arrest was induced in G0/G1 phase(P<0.05),and the expressions of cyclinD1,CDK4 and MMP7 proteins were decreased(P<0.05).Conclusion MNX1-AS1 is highly expressed in colorectal cancer tissues,and its expression level is related to TNM staging,lymph node metastasis and degrees of differentiation.The regulation of CyclinD1,CDK4 and MMP7 could be one of the mechanisms of MNX1-AS1 affecting the proliferation and metastasis of colorectal cancer cells.
作者
张维
张珂
蒋力
黄容海
鲁岩
赫嵘
李勤涛
李宝亮
贾哲
王杨
ZHANG Wei;ZHANG Ke;JIANG Li;HUANG Ronghai;LU Yan;HE Rong;LI Qintao;LI Baoliang;JIA Zhe;WANG Yang(Department of Hepatobiliary Surgery,Beijing Ditan Hospital,Capital Medical University, Beijing 100015,China)
出处
《标记免疫分析与临床》
CAS
2020年第5期833-838,共6页
Labeled Immunoassays and Clinical Medicine
