Human Cell

, Volume 32, Issue 2, pp 160–171 | Cite as

Suppression of miR-93-5p inhibits high-risk HPV-positive cervical cancer progression via targeting of BTG3

  • Jie Li
  • Zhao-Ping Chu
  • Hua Han
  • Yuan Zhang
  • Fei Tian
  • Jun-Qin Zhang
  • Xiang-Hua HuangEmail author
Research Article


This study explores the role of miR-93-5p in high-risk HPV-positive (HR-HPV) cervical cancer by targeting of BTG3. Cervical tissues were collected from 332 patients with conditions of chronic cervicitis (n = 42), low-grade cervical intraepithelial neoplasia (CIN I, n = 51), CIN II (n = 49), CIN III (n = 43), cervical cancer (n = 90), and normal cervical tissues (n = 57). HR-HPV DNA was detected by Hybrid Capture 2, and the expressions of miR-93-5p and BTG3 were determined by qRT-PCR and Western blot. The target relationship between miR-93-5p and BTG3 was verified by dual-luciferase reporter gene assay. HPV-positive cervical cancer cells (CaSki and HeLa) were divided into control, NC, inhibitor, BTG3, and mimic + BTG3 groups. CCK-8, Annexin V-APC/PI, and Transwell assays were applied to evaluate cell biological activities. MiR-93-5p was positively related but BTG3 was inversely related to HR-HPV infection. Additionally, miR-93-5p expression was negatively correlated with BTG3 expression in cervical cancer tissues infected with HR-HPV. HPV-positive cervical cancer cells showed higher miR-93-5p and lower BTG3 levels than negative cells. CaSki and HeLa cells in the inhibitor group showed increased BTG3 compared with the control group. After transfection with miR-93-5p inhibitor or BTG3 activation plasmid, proliferation and metastasis were inhibited, but apoptosis was promoted. The mimic + BTG3 group showed increased cell proliferation and metastasis but decreased cell apoptosis compared with the BTG3 group. Upregulated miR-93-5p was positively related but downregulated BTG3 was inversely related to HR-HPV infection, and inhibition of miR-93-5p may have blocked HPV-positive cervical cancer development by targeting of BTG3.


Cervical cancer High-risk HPV MiR-93-5p BTG3 



The authors are grateful for the many helpful suggestions and comments on this work.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interest.


  1. 1.
    Wu WJ, Shen Y, Sui J, et al. Integrated analysis of long noncoding RNA competing interactions revealed potential biomarkers in cervical cancer: Based on a public database. Mol Med Rep. 2018;17:7845–58.PubMedGoogle Scholar
  2. 2.
    Amarin ZO, Badria LF, Obeidat BR. Attitudes and beliefs about cervical smear testing in ever-married Jordanian women. East Mediterr Health J. 2008;14:389–97.PubMedGoogle Scholar
  3. 3.
    Song B, Ding C, Chen W, Sun H, Zhang M, Chen W. Incidence and mortality of cervical cancer in China, 2013. Chin J Cancer Res. 2017;29:471–76.CrossRefPubMedCentralPubMedGoogle Scholar
  4. 4.
    Bang HB, Lee YH, Lee YJ, Jeong KJ. High-level production of human papillomavirus (HPV) type 16 L1 in Escherichia coli. J Microbiol Biotechnol. 2016;26:356–63.CrossRefPubMedGoogle Scholar
  5. 5.
    Murdiyarso LS, Kartawinata M, Jenie I, et al. Single and multiple high-risk and low-risk Human Papillomavirus association with cervical lesions of 11,224 women in Jakarta. Cancer Causes Control. 2016;27:1371–79.CrossRefPubMedGoogle Scholar
  6. 6.
    Fakhreldin M, Elmasry K. Improving the performance of reflex Human Papilloma Virus (HPV) testing in triaging women with atypical squamous cells of undetermined significance (ASCUS): a retrospective study in a tertiary hospital in United Arab Emirates (UAE). Vaccine. 2016;34:823–30.CrossRefPubMedGoogle Scholar
  7. 7.
    Rennie W, Kanoria S, Liu C, et al. STarMirDB: a database of microRNA binding sites. RNA Biol. 2016;13:554–60.CrossRefPubMedCentralPubMedGoogle Scholar
  8. 8.
    Garbicz F, Mehlich D, Rak B, et al. Increased expression of the microRNA 106b ~ 25 cluster and its host gene MCM7 in corticotroph pituitary adenomas is associated with tumor invasion and Crooke’s cell morphology. Pituitary. 2017;20:450–63.CrossRefPubMedCentralPubMedGoogle Scholar
  9. 9.
    Lyu X, Fang W, Cai L, et al. TGFbetaR2 is a major target of miR-93 in nasopharyngeal carcinoma aggressiveness. Mol Cancer. 2014;13:51.CrossRefPubMedCentralPubMedGoogle Scholar
  10. 10.
    Zou J, Mi L, Yu XF, Dong J. Interaction of 14-3-3sigma with KCMF1 suppresses the proliferation and colony formation of human colon cancer stem cells. World J Gastroenterol. 2013;19:3770–80.CrossRefPubMedCentralPubMedGoogle Scholar
  11. 11.
    Zhu W, He J, Chen D, et al. Expression of miR-29c, miR-93, and miR-429 as potential biomarkers for detection of early stage non-small lung cancer. PLoS One. 2014;9:e87780.CrossRefPubMedCentralPubMedGoogle Scholar
  12. 12.
    Piccaluga PP, Navari M, De Falco G, et al. Virus-encoded microRNA contributes to the molecular profile of EBV-positive Burkitt lymphomas. Oncotarget. 2016;7:224–40.CrossRefPubMedGoogle Scholar
  13. 13.
    Zekri AN, Youssef AS, El-Desouky ED, et al. Serum microRNA panels as potential biomarkers for early detection of hepatocellular carcinoma on top of HCV infection. Tumour Biol. 2016;37:12273–86.CrossRefPubMedGoogle Scholar
  14. 14.
    Sharma S, Hussain S, Soni K, et al. Novel microRNA signatures in HPV-mediated cervical carcinogenesis in Indian women. Tumour Biol. 2016;37:4585–95.CrossRefPubMedGoogle Scholar
  15. 15.
    Namba-Fukuyo H, Funata S, Matsusaka K, et al. TET2 functions as a resistance factor against DNA methylation acquisition during Epstein–Barr virus infection. Oncotarget. 2016;7:81512–26.CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    Yen CS, Su ZR, Lee YP, Liu IT, Yen CJ. miR-106b promotes cancer progression in hepatitis B virus-associated hepatocellular carcinoma. World J Gastroenterol. 2016;22:5183–92.CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Ohta K, Hoshino H, Wang J, et al. MicroRNA-93 activates c-Met/PI3K/Akt pathway activity in hepatocellular carcinoma by directly inhibiting PTEN and CDKN1A. Oncotarget. 2015;6:3211–24.CrossRefPubMedGoogle Scholar
  18. 18.
    Gao D, Zhang Y, Zhu M, Liu S, Wang X. miRNA expression profiles of HPV-infected patients with cervical cancer in the Uyghur population in China. PLoS One. 2016;11:e0164701.CrossRefPubMedCentralPubMedGoogle Scholar
  19. 19.
    Cui H, Zhang S, Zhou H, Guo L. Direct downregulation of B-cell translocation gene 3 by microRNA-93 is required for desensitizing esophageal cancer to radiotherapy. Dig Dis Sci. 2017;62:1995–2003.CrossRefPubMedGoogle Scholar
  20. 20.
    Xiong Y, Sun F, Dong P, et al. iASPP induces EMT and cisplatin resistance in human cervical cancer through miR-20a-FBXL5/BTG3 signaling. J Exp Clin Cancer Res. 2017;36:48.CrossRefPubMedCentralPubMedGoogle Scholar
  21. 21.
    World Medical A. World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA. 2013;310:2191–4.CrossRefGoogle Scholar
  22. 22.
    Sargent A, Bailey A, Turner A, et al. Optimal threshold for a positive hybrid capture 2 test for detection of human papillomavirus: data from the ARTISTIC trial. J Clin Microbiol. 2010;48:554–8.CrossRefPubMedGoogle Scholar
  23. 23.
    Jovanovic M, Stefanoska I, Radojcic L, Vicovac L. Interleukin-8 (CXCL8) stimulates trophoblast cell migration and invasion by increasing levels of matrix metalloproteinase (MMP)2 and MMP9 and integrins alpha5 and beta1. Reproduction. 2010;139:789–98.CrossRefPubMedGoogle Scholar
  24. 24.
    Sathyapalan T, David R, Gooderham NJ, Atkin SL. Increased expression of circulating miRNA-93 in women with polycystic ovary syndrome may represent a novel, non-invasive biomarker for diagnosis. Sci Rep. 2015;5:16890.CrossRefPubMedCentralPubMedGoogle Scholar
  25. 25.
    Chen S, Chen X, Sun KX, et al. MicroRNA-93 promotes epithelial–mesenchymal transition of endometrial carcinoma cells. PLoS One. 2016;11:e0165776.CrossRefPubMedCentralPubMedGoogle Scholar
  26. 26.
    Wang L, Wang Q, Li HL, Han LY. Expression of MiR200a, miR93, metastasis-related gene RECK and MMP2/MMP9 in human cervical carcinoma–relationship with prognosis. Asian Pac J Cancer Prev. 2013;14:2113–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Zhao X, Cui Y, Jiang S, et al. Comparative study of HR HPV E6/E7 mRNA and HR-HPV DNA in cervical cancer screening. Zhonghua Yi Xue Za Zhi. 2014;94:3432–5.PubMedGoogle Scholar
  28. 28.
    Stiasny A, Kuhn C, Mayr D, et al. Immunohistochemical evaluation of E6/E7 HPV oncoproteins staining in cervical cancer. Anticancer Res. 2016;36:3195–8.PubMedGoogle Scholar
  29. 29.
    Tian Q, Li Y, Wang F, et al. MicroRNA detection in cervical exfoliated cells as a triage for human papillomavirus–positive women. J Natl Cancer Inst. 2014;106:1–8.CrossRefGoogle Scholar
  30. 30.
    Hassan T, Smith SG, Gaughan K, et al. Isolation and identification of cell-specific microRNAs targeting a messenger RNA using a biotinylated anti-sense oligonucleotide capture affinity technique. Nucleic Acids Res. 2013;41:e71.CrossRefPubMedCentralPubMedGoogle Scholar
  31. 31.
    Yoneda M, Suzuki T, Nakamura T, et al. Deficiency of antiproliferative family protein Ana correlates with development of lung adenocarcinoma. Cancer Sci. 2009;100:225–32.CrossRefPubMedGoogle Scholar
  32. 32.
    Chen X, Chen G, Cao X, Zhou Y, Yang T, Wei S. Downregulation of BTG3 in non-small cell lung cancer. Biochem Biophys Res Commun. 2013;437:173–8.CrossRefPubMedGoogle Scholar
  33. 33.
    Gou WF, Yang XF, Shen DF, et al. The roles of BTG3 expression in gastric cancer: a potential marker for carcinogenesis and a target molecule for gene therapy. Oncotarget. 2015;6:19841–67.PubMedCentralPubMedGoogle Scholar
  34. 34.
    Majid S, Dar AA, Ahmad AE, et al. BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer. Carcinogenesis. 2009;30:662–70.CrossRefPubMedCentralPubMedGoogle Scholar
  35. 35.
    Yu J, Zhang Y, Qi Z, Kurtycz D, Vacano G, Patterson D. Methylation-mediated downregulation of the B-cell translocation gene 3 (BTG3) in breast cancer cells. Gene Expr. 2008;14:173–82.PubMedGoogle Scholar
  36. 36.
    Ou YH, Chung PH, Hsu FF, Sun TP, Chang WY, Shieh SY. The candidate tumor suppressor BTG3 is a transcriptional target of p53 that inhibits E2F1. EMBO J. 2007;26:3968–80.CrossRefPubMedCentralPubMedGoogle Scholar
  37. 37.
    Zhang J, Wang F, Xu J, Wang X, Ye F, Xie X. Micro ribonucleic acid-93 promotes oncogenesis of cervical cancer by targeting RAB11 family interacting protein 1. J Obstet Gynaecol Res. 2016;42:1168–79.CrossRefPubMedGoogle Scholar
  38. 38.
    Shyamasundar S, Lim JP, Bay BH. miR-93 inhibits the invasive potential of triple-negative breast cancer cells in vitro via protein kinase WNK1. Int J Oncol. 2016;49:2629–36.CrossRefPubMedGoogle Scholar
  39. 39.
    Lv C, Wang H, Tong Y, et al. The function of BTG3 in colorectal cancer cells and its possible signaling pathway. J Cancer Res Clin Oncol. 2018;144:295–308.CrossRefPubMedGoogle Scholar
  40. 40.
    Wei K, Pan C, Yao G, et al. MiR-106b-5p promotes proliferation and inhibits apoptosis by regulating BTG3 in non-small cell lung cancer. Cell Physiol Biochem Int J Exp Cell Physiol Biochem Pharmacol. 2017; 44:1545–58.CrossRefGoogle Scholar
  41. 41.
    Yoshida Y, Hosoda E, Nakamura T, Yamamoto T. Association of ANA, a member of the antiproliferative Tob family proteins, with a Caf1 component of the CCR4 transcriptional regulatory complex. Jpn J Cancer Res. 2001;92:592–6.CrossRefPubMedCentralPubMedGoogle Scholar
  42. 42.
    Foda HD, Zucker S. Matrix metalloproteinases in cancer invasion, metastasis and angiogenesis. Drug Discov Today. 2001;6:478–82.CrossRefPubMedGoogle Scholar
  43. 43.
    Sugiura Y, Ma L, Sun B, et al. The plasminogen-plasminogen activator (PA) system in neuroblastoma: role of PA inhibitor-1 in metastasis. Cancer Res. 1999;59:1327–36.PubMedGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  • Jie Li
    • 1
    • 2
  • Zhao-Ping Chu
    • 2
  • Hua Han
    • 2
  • Yuan Zhang
    • 2
  • Fei Tian
    • 2
  • Jun-Qin Zhang
    • 2
  • Xiang-Hua Huang
    • 3
    Email author
  1. 1.Department of Obstetrics and GynecologyHebei Medical UniversityShijiazhuangChina
  2. 2.Department of GynecologyHebei General HospitalShijiazhuangChina
  3. 3.Department of GynecologySecond Hospital of Hebei Medical UniversityShijiazhuangChina

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