Volume 6 Issue 3
May  2015
Turn off MathJax
Article Contents
Tang Kaiwen, Tang Yunqiang, Gong Yuanfeng, et al. Effect of GPR49 gene on proliferation and invasive ability of hepatoma cells[J]. ORGAN TRANSPLANTATION, 2015, 6(3): 152-156. doi: 10.3969/j.issn.1674-7445.2015.03.005
Citation: Tang Kaiwen, Tang Yunqiang, Gong Yuanfeng, et al. Effect of GPR49 gene on proliferation and invasive ability of hepatoma cells[J]. ORGAN TRANSPLANTATION, 2015, 6(3): 152-156. doi: 10.3969/j.issn.1674-7445.2015.03.005

Effect of GPR49 gene on proliferation and invasive ability of hepatoma cells

doi: 10.3969/j.issn.1674-7445.2015.03.005
More Information
  • Corresponding author: Mai Cong, Email:maicong2010@126.com
  • Received Date: 2015-01-30
    Available Online: 2021-01-19
  • Publish Date: 2015-05-15
  •   Objective  To discuss the effect of G-protein-coupled receptor 49 (GPR49) gene on proliferation and invasive ability of hepatoma cell line Huh7 and its molecular biological mechanism.  Methods  According to the different transfected small interfering RNA(si-RNA), Huh7 cells were divided into the GPR49-siRNA(si-GPR49)group and the NC-siRNA (si-NC) group. Untransfected Huh7 cells were set as the control group. Messenger RNA(mRNA)and protein expression of GPR49, cyclin D1 and matrix metalloproteinase 9 (MMP9) in the cells of the three groups were respectively detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot method. The proliferation and invasive ability of the cells of each group were respectively detected by MTT method and Transwell method.  Results  The relative expression of GPR49 mRNA of the si-GPR49 group was (23.8±3.1)% of the control group (P < 0.05). Compared with the control group, the protein expression of GPR49, cyclin D1 and MMP9 of the si-GPR49 group decreased significantly(all in P < 0.05). The proliferation experiment results by MTT indicated that the optical density(OD)of the cells of the si-GPR49 group at 72 h was (0.53±0.12), which was significantly lower than that of the control group(1.35±0.28). The difference had statistical significance (P < 0.05). The average invaded cell counts of the si-GPR49 group were (13.6±2.5), which was significantly lower than (65.3±6.1) of the control group. The difference had statistical significance (P < 0.05).  Conclusions  GPR49-siRNA may inhibit the gene expression of GPR49 in Huh7 cells. Its mechanism may be that the proliferation of Huh7 cells is inhibited by reducing the level of cyclin D1; the migration and invasive ability of Huh7 cells is inhibited by affecting the expression level of MMP9.

     

  • loading
  • [1]
    Eng OS, Tsang AT, Moore D, et al. Outcomes of microwave ablation for colorectal cancer liver metastases:a single center experience[J]. J Surg Oncol, 2014, DOI: 10.1002/jso.23849.
    [2]
    Llovet JM. Liver cancer:time to evolve trial design after everolimus failure[J]. Nat Rev Clin Oncol, 2014, 11(9):506-507. doi: 10.1038/nrclinonc.2014.136
    [3]
    Huang Y, Yang X, Zhao F, et al. Overexpression of Dickkopf-1 predicts poor prognosis for patients with hepatocellular carcinoma after orthotopic liver transplantation by promoting cancer metastasis and recurrence[J]. Med Oncol, 2014, 31(7):966. doi: 10.1007/s12032-014-0966-8
    [4]
    Ge Y, Ding Y, Zhang J, et al. Effect of angiogenesis inhibitor SU6668 in combination with 5-Fu on liver metastasis from transplantation tumors of human colorectal cancer in nude mice[J]. Int J Clin Exp Med, 2014, 7(10):3578-3582. http://cn.bing.com/academic/profile?id=1452664954&encoded=0&v=paper_preview&mkt=zh-cn
    [5]
    Pu X, Guo QX, Long HA, et al. Effects of mTOR-STAT3 on the migration and invasion abilities of hepatoma cell and mTOR-STAT3 expression in liver cancer[J]. Asian Pac J Trop Med, 2014, 7(5):368-372. doi: 10.1016/S1995-7645(14)60057-2
    [6]
    Tanese K, Fukuma M, Yamada T, et al. G-protein-coupled receptor GPR49 is up-regulated in basal cell carcinoma and promotes cell proliferation and tumor formation[J]. Am J Pathol, 2008, 173(3):835-843. doi: 10.2353/ajpath.2008.071091
    [7]
    Rot S, Taubert H, Bache M, et al. A novel splice variant of the stem cell marker LGR5/GPR49 is correlated with the risk of tumor-related death in soft-tissue sarcoma patients[J]. BMC Cancer, 2011, 11:429. doi: 10.1186/1471-2407-11-429
    [8]
    Guo Y, Chen W, Wang W, et al. Simultaneous diagnosis and gene therapy of immuno-rejection in rat allogeneic heart transplantation model using a T-cell-targeted theranostic nanosystem[J]. ACS Nano, 2012, 6(12):10646-10657. doi: 10.1021/nn3037573
    [9]
    Shen WW, Zeng Z, Zhu WX, et al. MiR-142-3p functions as a tumor suppressor by targeting CD133, ABCG2, and Lgr5 in colon cancer cells[J]. J Mol Med, 2013, 91(8):989-1000. doi: 10.1007/s00109-013-1037-x
    [10]
    Guo Y, Liang X, Lu M, et al. Mammalian target of rapamycin as a novel target in the treatment of hepatocellular carcinoma[J]. Hepatogastroenterology, 2010, 57(101):913-918. https://www.ncbi.nlm.nih.gov/pubmed/21033251
    [11]
    Ye X, Guo Y, Zhang Q, et al. βKlotho suppresses tumor growth in hepatocellular carcinoma by regulating Akt/GSK-3β/cyclin D1 signaling pathway[J]. PLoS One, 2013, 8(1):e55615. doi: 10.1371/journal.pone.0055615
    [12]
    Chen Q, Cao HZ, Zheng PS. LGR5 promotes the proliferation and tumor formation of cervical cancer cells through the Wnt/β-catenin signaling pathway[J]. Oncotarget, 2014, 5(19):9092-9105. doi: 10.18632/oncotarget
    [13]
    da Silva-Diz V, Solé-Sánchez S, Valdés-Gutiérrez A, et al. Progeny of Lgr5-expressing hair follicle stem cell contributes to papillomavirus-induced tumor development in epidermis[J]. Oncogene, 2013, 32(32):3732-3743. doi: 10.1038/onc.2012.375
    [14]
    Fafilek B, Krausova M, Vojtechova M, et al. Troy, a tumor necrosis factor receptor family member, interacts with lgr5 to inhibit wnt signaling in intestinal stem cells[J]. Gastroenterology, 2013, 144(2):381-391. doi: 10.1053/j.gastro.2012.10.048
    [15]
    Gao FJ, Chen JY, Wu HY, et al. Lgr5 over-expression is positively related to the tumor progression and HER2 expression in stage pTNM IV colorectal cancer[J]. Int J Clin Exp Pathol, 2014, 7(4):1572-1579. http://cn.bing.com/academic/profile?id=291580039&encoded=0&v=paper_preview&mkt=zh-cn
    [16]
    Kobayashi S, Yamada-Okabe H, Suzuki M, et al. LGR5-positive colon cancer stem cells interconvert with drug-resistant LGR5-negative cells and are capable of tumor reconstitution[J]. Stem Cells, 2012, 30(12):2631-2644. doi: 10.1002/stem.v30.12
    [17]
    Spengler RM, Oakley CK, Davidson BL. Functional microRNAs and target sites are created by lineage-specific transposition[J]. Hum Mol Genet, 2014, 23(7):1783-1793. doi: 10.1093/hmg/ddt569
    [18]
    Zhou MK, Liu XJ, Zhao ZG, et al. MicroRNA-100 functions as a tumor suppressor by inhibiting Lgr5 expression in colon cancer cells[J]. Mol Med Rep, 2015, 11(4):2947-2952. doi: 10.3892/mmr.2014.3052/download
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(2)

    Article Metrics

    Article views (80) PDF downloads(4) Cited by()
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return