miR-24靶向调控CARMA3抑制人结肠癌细胞系SW620增殖
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摘要
目的探讨miR-24对人结肠癌SW620细胞CARMA3(含半胱天冬酶募集结构域的膜相关乌苷酸激酶蛋白3)的靶向调控作用及对细胞增殖的影响。方法培养SW620细胞,用生物信息学预测miR-24和CARMA3的靶向匹配关系,并用双荧光素酶报告基因系统鉴定。Lipo3000转染miR-24模拟物后,real-time PCR检测miR-24和CARMA3 m RNA表达; Western blot检测CARMA3蛋白表达; CCK8法检测SW620细胞增殖。结果 miR-24和CARMA3匹配良好,miR-24能够结合CARMA3 m RNA 3'UTR,并有效抑制其表达(P <0. 05)。miR-24能够负向调控CARMA3表达和SW620细胞增殖(P<0. 05)。结论 miR-24通过靶向作用于CARMA3 m RNA 3'UTR能够负向调控CARMA3基因的表达及负向调控SW620细胞的增殖。
Objective To identify the role of miR-24 in human colorectal cancer SW620 cell proliferation. Methods SW620 cells were cultured in vitro. The miR-24 was predicted with bioinformatics and identified with dual luciferase report system. Expression of miR-24 and CARMA3 was determined with real-time PCR and Western blot after transfection of miR-24. The proliferation of SW620 cells was detected in vitro by CCK8 assay. Results miR-24 can inhibit CARMA3 expression by binding to 3'UTR of CARMA3 mRNA. miR-24 can negatively control the CARMA3 expression. The proliferation of SW620 cells was negatively controlled by miR-24. Conclusions The miR-24 may negatively regulate CARMA3 expression by binding to its 3 ' UTR in human colorectal cancer SW620 cells and cell proliferation.
Objective To identify the role of miR-24 in human colorectal cancer SW620 cell proliferation. Methods SW620 cells were cultured in vitro. The miR-24 was predicted with bioinformatics and identified with dual luciferase report system. Expression of miR-24 and CARMA3 was determined with real-time PCR and Western blot after transfection of miR-24. The proliferation of SW620 cells was detected in vitro by CCK8 assay. Results miR-24 can inhibit CARMA3 expression by binding to 3'UTR of CARMA3 mRNA. miR-24 can negatively control the CARMA3 expression. The proliferation of SW620 cells was negatively controlled by miR-24. Conclusions The miR-24 may negatively regulate CARMA3 expression by binding to its 3 ' UTR in human colorectal cancer SW620 cells and cell proliferation.
引文
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[2]Scudiero I,Vito P,Stilo R.The three CARMA sisters:so different,so similar:a portrait of the three CARMA proteins and their involvement in human disorders[J].J Cell Physiol,2014,229:990-997.
[3]Jiang C,Zhou Z,Quan Y,et al.CARMA3 is a host factor regulating the balance of inflammatory and antiviral responses against viral infection[J].Cell Rep,2016,14:2389-2401.
[4]Cimmino A,Calin GA,Fabbri M,et al.mi R-15 and mi R-16 induce apoptosis by targeting BCL2[J].Proc Natl Acad Sci U S A,2005,102:13944-13949.
[5]Birnie KA,Prele CM,Thompson PJ,et al.Targeting micro RNA to improve diagnostic and therapeutic approaches for malignant mesothelioma[J].Oncotarget,2017,8:78193-78207.
[6]Juilland M,Thome M.Role of the CARMA1/BCL10/MALT1 complex in lymphoid malignancies[J].Curr Opin Hematol,2016,23:402-409.
[7]Bertin J,Wang L,Guo Y,et al.CARD11 and CARD14are novel caspase recruitment domain(CARD)/membrane-associated guanylate kinase(MAGUK)family members that interact with BCL10 and activate NF-kappa B[J].J Biol Chem,2001,276:11877-11882.
[8]Xia ZX,Li ZX,Zhang M,et al.CARMA3 regulates the invasion,migration,and apoptosis of non-small cell lung cancer cells by activating NF-small ka,Cyrillic B and suppressing the P38 MAPK signaling pathway[J].Exp Mol Pathol,2016,100:353-360.
[9]Mazzone P,Scudiero I,Ferravante A,et al.Functional characterization of zebrafish(Danio rerio)Bcl10[J].PLoS One,2015,10:e122365.doi:10.1371/journal.pone.0122365.
[10]Zhang S,Pan D,Jia XM,et al.The CARMA3-BCL10-MALT1(CBM)complex contributes to DNA damage-induced NF-kappaB activation and cell survival[J].Protein Cell,2017,8:856-860.
[11]Sun J.CARMA3:A novel scaffold protein in regulation of NF-kappaB activation and diseases[J].World J Biol Chem,2010,1:353-361.
[12]Jiang C,Zhu Y,Zhou Z,et al.TMEM43/LUMA is a key signaling component mediating EGFR-induced NF-kappaBactivation and tumor progression[J].Oncogene,2017,36:2813-2823.