长链非编码RNA SNHG16通过miR-141-3p/FOXJ3通路调控肝细胞癌的增殖

摘要/Abstract

摘要： 目的检测长链非编码RNA（lncRNA）小核仁RNA宿主基因16（SNHG16）在肝细胞癌中的表达特征,观察其通过微小RNA141-3p（miR-141-3p）/叉头转录因子J3（FOXJ3）对肝癌细胞增殖的调控作用。方法应用生物信息技术预测SNHG16和miR-141-3p、miR-141-3p和FOXJ3可能的结合位点。收集肝细胞癌患者共49例,留取术后肿瘤组织和距肿物边缘>3cm的癌旁肝组织。选择人正常肝细胞株HL-7702,并选择人肝癌细胞株Hep-3B和Huh7,构建转染pc-DNA3.1-SNHG16的pc-DNA3.1-SNHG16组、转染pc-DNA3.1-SNHG16-siRNA的si-SNHG16组,转染miR-141-3p inhibitor的miR-141-3p inhibitor组、转染miR-141-3p mimic的miR-141-3p mimic组、转染pc-DNA3.1-FOXJ3-siRNA的si-FOXJ3组。应用CCK-8检测细胞增殖活性,实时荧光定量PCR法（qRT-PCR）检测SNHG16和miR-141-3p的表达,免疫组化法检测FOXJ3和Ki67的表达,WB检测FOXJ3的表达;应用双荧光素酶报告基因实验验证SNHG16和miR-141-3p、miR-141-3p和FOXJ3的靶向关系。结果生物信息分析显示,SNHG16和miR-141-3p、miR-141-3p和FOXJ3具有可能结合的碱基序列;与正常肝细胞株比较,人肝癌细胞株中SNHG16和FOXJ3高表达,miR-141-3p低表达;与空白对照组和空载体转染组比较,pc-DNA3.1-SNHG16组细胞增殖活性在48h时明显升高,si-SNHG16组在48h明显降低,均持续至96h;双荧光素酶报告基因实验显示,SNHG16和miR-141-3p、miR-141-3p和FOXJ3具有靶向关系;挽救实验显示,沉默miR-141-3p可部分逆转沉默SNHG16和FOXJ3对细胞增殖的抑制作用;肝细胞癌组织中SNHG16的表达量和FOXJ3表达的阳性率均明显高于癌旁肝组织,miR-141-3p的表达量明显低于癌旁肝组织;肝细胞癌中SNHG16和miR-141-3p、miR-141-3p和FOXJ3均呈负相关性,FOXJ3和Ki67呈正相关性。结论肝细胞癌中SNHG16的表达升高,SNHG16通过miR-141-3p/FOXJ3途径促进肝癌的增殖。

关键词: 肝细胞癌, 长链非编码RNA SNHG16, miR-141-3p, FOXJ3, 增殖

Abstract: Objective To detect the expression characteristics of lncRNA SNHG16 in hepatocellular carcinoma, and observe the proliferation regulatory effect by miR-141-3p/FOXJ3 on the hepatocarcinoma cells. Methods Potential binding sites were predicted between SNHG16 and miR-141-3p, miR-141-3p and FOXJ3 by bioinformatics analyses. Forty-nine cases of hepatocellular carcinoma were collected, the tumor tissue and the adjacent liver tissue were retained. The normal human hepatocyte HL-7702, hepatocarcinoma cells of Hep-3B and Huh7 were selected. The pc-DNA3.1-SNHG16 group, si-SNHG16 group, miR-141-3p inhibitor group, miR-141-3p mimic group, and si-FOXJ3 group were constructed The cell proliferation activity was detected through CCK-8 method. The expression of SNHG16 and miR-141-3p was detected by real-time fluorescence quantitative PCR(qRT-PCR), FOXJ3 and Ki67 were detected through immunohistochemistry method, FOXJ3 were detected by Western blot. Target relationship was verified between SNHG16 and miR-141-3p, miR-141-3p and FOXJ3 by double luciferase reporter gene experiment. Results Possible combination of base sequences were found between SNHG16 and miR-141-3p, miR-141-3p and FOXJ3 by bioinformatics analysis. The expression of SNHG16 and FOXJ3 were higher in human hepatoma cell line than in normal hepatocyte line. The expression of miR-141-3p was lower in human hepatoma cell line than that in normal hepatocyte line. Compared with the control group and the empty vector transfection group, the proliferation activity was increased at 48h in pc-DNA3.1-SNHG16 group, while it was decreased in si-SNHG16 group, which lasted to 96h. Target relationship was found between SNHG16 and miR-141-3p, miR-141-3p and FOXJ3 in double luciferase reporter gene test. The rescue experiment showed that the inhibition of miR-141-3p could partially reverse the inhibition effect of si-SNHG16 and si-FOXJ3 on cell proliferation. The expression of SNGHN16 and the positive rate of FOXJ3 in hepatocellular carcinoma were significantly higher than that in adjacent liver tissues, and the expression of miR-141-3p was significantly lower than that in adjacent liver tissues. SNHG16 was negatively correlated with miR-141-3p, miR-141-3p and FOXJ3, and positively correlated with FOXJ3 and Ki67 in hepatocellular carcinoma. Conclusion Expression of SNHG16 is increased in hepatocellular carcinoma, which promotes the proliferation through miR-141-3p/FOXJ3 pathway.

Key words: hepatocellular carcinoma, lnc SNHG16, miR-141-3p, FOXJ3, proliferation

中图分类号:

R735

吴涛, 杨柳, 聂山茂, 张杨, 张鑫. 长链非编码RNA SNHG16通过miR-141-3p/FOXJ3通路调控肝细胞癌的增殖[J]. 承德医学院学报, 2022, 39(4): 271-276.

WU Tao, YANG Liu, NIE Shan-mao, ZHANG Yang, ZHANG Xin. Proliferation Regulation of LncRNA SNHG16 through miR-141-3p/FOXJ3 Pathway in Hepatocellular Carcinoma[J]. Journal of Chengde Medical University, 2022, 39(4): 271-276.

参考文献

[1] Zheng X, Lin J, Wu H, et al.Forkhead box (FOX) G1 promotes hepatocellular carcinoma epithelial-Mesenchymal transition by activating Wnt signal through forming T-cell factor-4/Beta-catenin/FOXG1 complex[J]. J Exp Clin Cancer Res, 2019, 38(1): 475-477.
[2] Luo Q, Wang CQ, Yang LY, et al.FOXQ1/NDRG1 axis exacerbates hepatocellular carcinoma initiation via enhancing crosstalk between fibroblasts and tumor cells[J]. Cancer Lett, 2018, 28: 21-34.
[3] Chen J, Huang X, Wang W, et al.LncRNA CDKN2BAS predicts poor prognosis in patients with hepatocellular carcinoma and promotes metastasis via the miR-153-5p/ARHGAP18 signaling axis[J]. Aging (Albany NY), 2018, 10(11): 3371-3381.
[4] Fu Y, Li Y, Wang X, et al.Overexpression of miR-425-5p is associated with poor prognosis and tumor progression in non-small cell lung cancer[J]. Cancer Biomark, 2020, 27(2): 147-156.
[5] Yu Y, Chen F, Yang Y, et al.lncRNA SNHG16 is associated with proliferation and poor prognosis of pediatric neuroblastoma[J]. Int J Oncol, 2019, 55(1): 93-102.
[6] Cheng W, Hao CY, Zhao S, et al.SNHG16 promotes the progression of osteoarthritis through activating microRNA-93-5p/CCND1 axis[J]. Eur Rev Med Pharmacol Sci, 2019, 23(21): 9222-9229.
[7] Gong CY, Tang R, Nan W, et al.Role of SNHG16 in human cancer[J]. Clin Chim Acta, 2020, 503: 175-180.
[8] Jeon HJ, Lee JG, Kim K, et al.Peripheral blood transcriptome analysis and development of classification model for diagnosing antibody-mediated rejection vs accommodation in ABO-incompatible kidney transplant[J]. Am J Transplant, 2020, 20(1): 112-124.
[9] Jin J, Zhou S, Li C, et al.MiR-517a-3p accelerates lung cancer cell proliferation and invasion through inhibiting FOXJ3 expression[J]. Life Sci, 2014,108(1): 48-53.
[10] De la Rocha AMA, González-Huarriz M, Guruceaga E, et al. miR-425-5p, a SOX2 target, regulates the expression of FOXJ3 and RAB31 and promotes the survival of GSCs[J]. Arch Clin Biomed Res, 2020, 4(3): 221-238.
[11] Yang M, Wei W.SNHG16: A Novel Long-Non Coding RNA in Human Cancers[J]. Onco Targets Ther, 2019, 31(12): 11679-11690.
[12] Guo JQ, Yang ZJ, Wang S, et al.LncRNA SNHG16 functions as an oncogene by sponging miR-200a-3p in pancreatic cancer[J]. Eur Rev Med Pharmacol Sci, 2020, 24(13): 7218-7219.
[13] Liao S, Xing S, Ma Y.LncRNA SNHG16 sponges miR-98-5p to regulate cellular processes in osteosarcoma[J]. Cancer Chemother Pharmacol, 2019, 83(6): 1065-1074.
[14] Li M, Huang H, Cheng F, et al.miR-141-3p promotes proliferation and metastasis of nasopharyngeal carcinoma by targeting NME1[J]. Adv Med Sci, 2020, 65(2): 252-258.
[15] Li W, Cui Y, Wang D, et al.MiR-141-3p functions as a tumor suppressor through directly targeting ZFR in non-small cell lung cancer[J].Biochem Biophys Res Commun, 2019, 509(3): 647-656.
[16] Wang N, Li P, Liu W, et al.miR-141-3p suppresses proliferation and promotes apoptosis by targeting GLI2 in osteosarcoma cells[J]. Oncol Rep, 2018, 39(2): 747-754.
[17] Guo JQ, Yang ZJ, Wang S, et al.LncRNA SNHG16 functions as an oncogene by sponging miR-200a-3p in pancreatic cancer[J]. Eur Rev Med Pharmacol Sci, 2020, 24(4): 1718-1724.
[18] Chen H, Li M, Huang P.LncRNA SNHG16 Promotes Hepatocellular Carcinoma Proliferation, Migration and Invasion by Regulating miR-186 Expression[J]. J Cancer, 2019, 10(15): 3571-3581.
[19] Ye J, Tan L, Fu Y, et al.LncRNA SNHG15 promotes hepatocellular carcinoma progression by sponging miR-141-3p[J]. J Cell Biochem, 2019, 120(12): 19775-19783.
[20] Katoh M, Katoh M.Human FOX gene family (Review)[J]. Int J Oncol, 2004, 25(5): 1495-1500.
[21] Zhang JY, Su XP, Li YN, et al.MicroRNA-425-5p promotes the development of prostate cancer via targeting forkhead box J3[J]. Eur Rev Med Pharmacol Sci, 2019, 23(2): 547-554.
[22] Lim LJ, Wong SYS, Huang F, et al.Roles and Regulation of Long Noncoding RNAs in Hepatocellular Carcinoma[J]. Cancer Res, 2019, 79(20): 5131-5139.
[23] Cui XP, Wang CX, Wang ZY, et al. LncRNA TP73-AS1 sponges miR-141-3p to promote the migration and invasion of pancreatic cancer cells through the up-regulation of BDH2[J]. Biosci Rep, 2019, 39(3): BSR20181937.