miR-122通过靶定LMNB2基因抑制肝癌细胞活性的研究
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摘要
目的探讨miR-122在肝癌进展中的作用及其机制。方法应用荧光素酶报告基因实验和Western blot实验证实miR-122对LMNB2基因的调控作用。应用菌落形成试验和Transwell细胞侵袭实验分析LMNB2基因对肝癌细胞生物活性的影响。结果野生型转染miR-122 mimics后荧光素酶活性弱于对照组(P<0.05);突变体转染miR-122 mimics后荧光素酶活性与对照组差异无统计学意义(P>0.05)。在肝癌细胞Hep3B中沉默miR-122表达,LMNB2表达水平明显高于对照组(P<0.05)。在肝癌细胞SMMC7721中过表达miR-122后LMNB2表达水平明显低于对照组(P<0.05)。在肝癌细胞Hep3B中过表达LMNB2,肝癌细胞的侵袭及迁移能力明显强于对照组(P<0.05);在肝癌细胞SMMC7721中沉默LMNB2表达,肝癌细胞的侵袭及迁移能力明显低于对照组(P<0.05)。在肝癌细胞Hep3B中过表达LMNB2,肝癌细胞的增殖能力明显强于对照组(P<0.05);在肝癌细胞SMMC7721中沉默LMNB2表达,肝癌细胞的增殖能力明显低于对照组(P<0.05)。结论 miR-122可以通过靶定LMNB2抑制肝癌细胞的生物活性,其可以作为肝细胞肝癌早期诊断指标和治疗的靶点。
Objective To investigate the role and mechanism of miR-122 in the progression of liver cancer. Methods The regulatory effect of miR-122 on LMNB2 gene was confirmed by luciferase reporter gene assay and Western blot. Then the effect of LMNB2 gene on the bioactivity of hepatocellular carcinoma(HCC) cells was analyzed by colony formation assay and transwell cell invasion assay.Results The luciferase activity of the wild-type mir-122 mimics was lower than that of the control group(P<0.05). The luciferase activity of the mutant transfected with mir-122 mimics was not significantly different from that of the control group(P>0.05). The expression of mir-122 was silenced in hepatocellular carcinoma cells Hep3B, and the expression level of LMNB2 was significantly higher than that of the control group(P<0.05). LMNB2 expression level was significantly lower than that of the control group after overexpression of mir-122 in liver cancer SMMC7721 cells(P<0.05). LMNB2 was overexpressed in hepatocellular carcinoma cells Hep3B, and the invasion and migration ability of hepatocellular carcinoma cells was significantly stronger than that of the control group(P<0.05). The LMNB2 expression was silenced in SMMC7721 HCC cells, and the invasion and migration ability of HCC cells was significantly lower than that of the control group(P<0.05). LMNB2 was overexpressed in hepatocellular carcinoma cells Hep3B, and the proliferation capacity of hepatocellular carcinoma cells was significantly stronger than that of the control group(P<0.05). The LMNB2 expression was silenced in SMMC7721 hepatocellular carcinoma cells, and the proliferation capacity of hepatocellular carcinoma cells was significantly lower than that of the control group(P<0.05). Conclusion miR-122 can inhibit the biological activity of HCC cells by targeting LMNB2, which can be used as an early diagnostic indicator and therapeutic target for HCC.
Objective To investigate the role and mechanism of miR-122 in the progression of liver cancer. Methods The regulatory effect of miR-122 on LMNB2 gene was confirmed by luciferase reporter gene assay and Western blot. Then the effect of LMNB2 gene on the bioactivity of hepatocellular carcinoma(HCC) cells was analyzed by colony formation assay and transwell cell invasion assay.Results The luciferase activity of the wild-type mir-122 mimics was lower than that of the control group(P<0.05). The luciferase activity of the mutant transfected with mir-122 mimics was not significantly different from that of the control group(P>0.05). The expression of mir-122 was silenced in hepatocellular carcinoma cells Hep3B, and the expression level of LMNB2 was significantly higher than that of the control group(P<0.05). LMNB2 expression level was significantly lower than that of the control group after overexpression of mir-122 in liver cancer SMMC7721 cells(P<0.05). LMNB2 was overexpressed in hepatocellular carcinoma cells Hep3B, and the invasion and migration ability of hepatocellular carcinoma cells was significantly stronger than that of the control group(P<0.05). The LMNB2 expression was silenced in SMMC7721 HCC cells, and the invasion and migration ability of HCC cells was significantly lower than that of the control group(P<0.05). LMNB2 was overexpressed in hepatocellular carcinoma cells Hep3B, and the proliferation capacity of hepatocellular carcinoma cells was significantly stronger than that of the control group(P<0.05). The LMNB2 expression was silenced in SMMC7721 hepatocellular carcinoma cells, and the proliferation capacity of hepatocellular carcinoma cells was significantly lower than that of the control group(P<0.05). Conclusion miR-122 can inhibit the biological activity of HCC cells by targeting LMNB2, which can be used as an early diagnostic indicator and therapeutic target for HCC.
引文
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[11] Yan Y,Li C,Sun B,et al.DCAF1 is involved in HCV replication through regulation of miR-122[J].Arch Virol,2018,163(4):977-985.
[2] Cheng C,Xiaohua W,Ning J,et al.MiR-122 exerts anti-proliferative and apoptotic effects on nasopharyngeal carcinoma cells via the PI3K/AKT signaling pathway[J].Cell Mol Biol(Noisy-le-grand),2017,64(13):21-25.
[3] Duan Y,Dong Y,Dang R,et al.MiR-122 inhibits epithelial mesenchymal transition by regulating P4HA1 in ovarian cancer cells[J].Cell Biol Int,2018,42(11):1564-1574.
[4] Guo L,Yin M,Wang Y,et al.CREB1,a direct target of miR-122,promotes cell proliferation and invasion in bladder cancer[J].Oncol Lett,2018,16(3):3842-3848.
[5] Xu Z,Liu G,Zhang M,et al.miR-122-5p Inhibits the Proliferation,Invasion and Growth of Bile Duct Carcinoma Cells by Targeting ALDOA[J].Cell Physiol Biochem,2018,48(6):2596-2606.
[6] Howell LS,Ireland L,Park BK,et al.MiR-122 and other microRNAs as potential circulating biomarkers of drug-induced liver injury[J].Expert Rev Mol Diagn,2018,18(1):47-54.
[7] Baranova A,Maltseva D,Tonevitsky A,et al.Adipose may actively delay progression of NAFLD by releasing tumor-suppressing,anti-fibroticmiR-122 into circulation[J].Obes Rev,2019,20(1):108-118.
[8] Akuta N,Suzuki F,Kobayashi M,et al.Circulating microRNA-122 levels are important predictor of hepatitis B virus surface antigen seroclearance[J].J Med Virol,2018,90(10):1586-1592.
[9] Amador-Caňizares Y,Panigrahi M,Huys A,et al.miR-122,small RNA annealing and sequence mutations alter the predicted structure of the Hepatitis C virus 5′ UTR RNA to stabilize and promote viral RNA accumulation[J].Nucleic Acids Res,2018,46(18):9776-9792.
[10] Schult P,Roth H,Adams RL,et al.microRNA-122 amplifies hepatitis C virus translation by shaping the structure of the internal ribosomal entry site[J].Nat Commun,2018,9(1):2613.
[11] Yan Y,Li C,Sun B,et al.DCAF1 is involved in HCV replication through regulation of miR-122[J].Arch Virol,2018,163(4):977-985.