Hsa_circ_0006427 Suppresses Multiplication, Migration and Invasion of Non-Small Cell Lung Cancer Cells through miR-346/VGLL4 Pathway

Document Type : Original Article


1 Department of Thoracic Surgery, Pinghu First People's Hospital, Pinghu City, Jiaxing, Zhejiang, China

2 Department of Anesthesiology, Pinghu First People's Hospital, Pinghu City, Jiaxing, Zhejiang, China

3 Department of Thoracic Surgery, Zhejiang Hospital, Xihu District, Hangzhou, Zhejiang, China


Objective: Circular RNAs (circRNAs) are identified as key modulators in cancer biology. Nonetheless, the role of circ_0006427 in non-small cell lung cancer (NSCLC) and its modulatory mechanism are undefined. This study aimed to investigate the potential function and mechanism of circ_0006427 in NSCLC.
Materials and Methods: In this experimental study, circ_0006427, miR-346 and vestigial like family member 4 (VGLL4) mRNA expressions were analyzed in NSCLC tissues and cells, using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Multiplication, migration and invasion of NSCLC cells were examined using the CCK-8 method and Transwell experiment, respectively. Dual-luciferase reporter gene experiments were conducted to identify the paring relationship between circ_0006427 and miR-346. Western blot was employed to determine expressions of VGLL4 and epithelial-mesenchymal transition (EMT) markers on protein levels. Immuno histochemistry (IHC) method was adopted to assess VGLL4 protein expression in NSCLC tissues.

Results: Circ_0006427 expression was down-regulated in NSCLC tissues and cells, and circ_0006427 suppressed multiplication, migration, invasion and EMT of NSCLC cells. miR-346 expression was upregulated in NSCLC tissues and cells, and miR-346 worked as a target of circ_0006427. VGLL4 was down-regulated in NSCLC tissues and cells, and knockdown of VGLL4 accelerated multiplication, migration, invasion and EMT of NSCLC cells. Circ_0006427 enhanced VGLL4 expression by competitively binding with miR-346.
Conclusion: Circ_0006427/miR-346/VGLL4 axis regulated NSCLC progression.


  1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018; 68(6): 394-424.
  2. Radovic M, Kanesvaran R, Rittmeyer A, Früh M, Minervini F, Glatzer M, et al. Multidisciplinary treatment of lung cancer in older patients: a review. J Geriatr Oncol. 2019; 10(3): 405-410.
  3. Testa U, Castelli G, Pelosi E. Lung cancers: molecular characterization, clonal heterogeneity and evolution, and cancer stem cells. Cancers (Basel). 2018; 10(8): 248.
  4. Meng S, Zhou H, Feng Z, Xu Z, Tang Y, Li P, et al. CircRNA: functions and properties of a novel potential biomarker for cancer. Mol Cancer. 2017; 16(1): 94.
  5. Kulcheski FR, Christoff AP, Margis R. Circular RNAs are miRNA sponges and can be used as a new class of biomarker. J Biotechnol. 2016; 238: 42-51.
  6. Liu T, Ye P, Ye Y, Lu S, Han B. Circular RNA hsa_circRNA_002178 silencing retards breast cancer progression via microRNA-328-3pmediated inhibition of COL1A1. J Cell Mol Med. 2020; 24(3): 2189- 2201.
  7. Li X, Wang J, Zhang C, Lin C, Zhang J, Zhang W, et al. Circular RNA circITGA7 inhibits colorectal cancer growth and metastasis by modulating the Ras pathway and upregulating transcription of its host gene ITGA7. J Pathol. 2018; 246(2): 166-179.
  8. AbouHaidar MG, Venkataraman S, Golshani A, Liu B, Ahmad T. Novel coding, translation, and gene expression of a replicating covalently closed circular RNA of 220 nt. Proc Natl Acad Sci USA. 2014; 111(40): 14542-14547.
  9. Xu Q, Deng B, Li M, Chen Y, Zhuan L. circRNA-UBAP2 promotes the proliferation and inhibits apoptosis of ovarian cancer though miR-382-5p/PRPF8 axis. J Ovarian Res. 2020; 13(1): 81.
  10. Wang K, Sun Y, Tao W, Fei X, Chang C. Androgen receptor (AR) promotes clear cell renal cell carcinoma (ccRCC) migration and invasion via altering the circHIAT1/miR-195-5p/29a-3p/29c-3p/ CDC42 signals. Cancer Lett. 2017; 394: 1-12.
  11. Fan L, Cao Q, Liu J, Zhang J, Li B. Circular RNA profiling and its potential for esophageal squamous cell cancer diagnosis and prognosis. Mol Cancer. 2019; 18(1): 16.
  12. Yao Y, Hua Q, Zhou Y. CircRNA has_circ_0006427 suppresses the progression of lung adenocarcinoma by regulating miR-6783-3p/ DKK1 axis and inactivating Wnt/β-catenin signaling pathway. Biochem Biophys Res Commun. 2019; 508(1): 37-45.
  13. Thomson DW, Dinger ME. Endogenous microRNA sponges: evidence and controversy. Nat Rev Genet. 2016; 17(5): 272-283.
  14. Long Z, Wang Y. miR-195-5p suppresses lung cancer cell proliferation, migration, and invasion Via FOXK1. Technol Cancer Res Treat. 2020; 19: 1533033820922587.
  15. Guo Z, Li J, Sun J, Sun L, Zhou Y, Yu Z. miR-346 Promotes HCC progression by suppressing breast cancer metastasis suppressor 1 expression. Oncol Res. 2018; 26(7): 1073-1081.
  16. Sun CC, Li SJ, Yuan ZP, Li DJ. MicroRNA-346 facilitates cell growth and metastasis, and suppresses cell apoptosis in human non-small cell lung cancer by regulation of XPC/ERK/Snail/E-cadherin pathway. Aging (Albany NY). 2016; 8(10): 2509-2524.
  17. Lin Z, Guo H, Cao Y, Zohrabian S, Zhou P, Ma Q, et al. Acetylation of VGLL4 regulates hippo-YAP signaling and postnatal cardiac growth. Dev Cell. 2016; 39(4): 466-479.
  18. Zhang W, Gao Y, Li P, Shi Z, Guo T, Li F, et al. VGLL4 functions as a new tumor suppressor in lung cancer by negatively regulating the YAP-TEAD transcriptional complex. Cell Res. 2014; 24(3): 331-343.
  19. Zhou J, Wang H, Chu J, Huang Q, Li G, Yan Y, et al. Circular RNA hsa_circ_0008344 regulates glioblastoma cell proliferation, migration, invasion, and apoptosis. J Clin Lab Anal. 2018; 32(7): e22454.
  20. Li X, Yang L, Chen LL. The biogenesis, functions, and challenges of circular RNAs. Mol Cell. 2018; 71(3): 428-442.
  21. Dong Y, Xu T, Zhong S, Wang B, Zhang H, Wang X, et al. Circ_0076305 regulates cisplatin resistance of non-small cell lung cancer via positively modulating STAT3 by sponging miR-296-5p. Life Sci. 2019; 239: 116984.
  22. Wang L, Tong X, Zhou Z, Wang S, Lei Z, Zhang T, et al. Circular RNA hsa_circ_0008305 (circPTK2) inhibits TGF-β-induced epithelial- mesenchymal transition and metastasis by controlling TIF1γ in non-small cell lung cancer. Mol Cancer. 2018; 17(1): 140.
  23. Wang L, Tong X, Zhou Z, Wang S, Lei Z, Zhang T, et al. Circular RNA hsa_circ_0008305 (circPTK2) inhibits TGF-β-induced epithelial- mesenchymal transition and metastasis by controlling TIF1γ in non-small cell lung cancer. Mol Cancer. 2018; 17(1): 140.
  24. Liang G, Meng W, Huang X, Zhu W, Yin C, Wang C, et al. miR-196b-5p-mediated downregulation of TSPAN12 and GATA6 promotes tumor progression in non-small cell lung cancer. Proc Natl Acad Sci U S A. 2020; 117(8): 4347-4357.
  25. Khandelwal A, Seam RK, Gupta M, Rana MK, Prakash H, Vasquez KM, et al. Circulating microRNA-590-5p functions as a liquid biopsy marker in non-small cell lung cancer. Cancer Sci. 2020; 111(3): 826-839.
  26. Li X, Chen SH, Zeng JW. MiR-421 is overexpressed and promotes cell proliferation in non-small cell lung cancer. Med Princ Pract. 2020; 29(1): 80-89.
  27. Kim DH, Park S, Kim H, Choi YJ, Kim SY, Sung KJ, et al. Tumorderived exosomal miR-619-5p promotes tumor angiogenesis and metastasis through the inhibition of RCAN1.4. Cancer Lett. 2020; 475: 2-13.
  28. Wang Y, Xu R, Zhang D, Lu T, Yu W, Wo Y, et al. Circ-ZKSCAN1 regulates FAM83A expression and inactivates MAPK signaling by targeting miR-330-5p to promote non-small cell lung cancer progression. Transl Lung Cancer Res. 2019; 8(6): 862-875.
  29. Li S, Niu X, Li H, Liang Y, Sun Z, Yan Y. Circ_0000003 promotes the proliferation and metastasis of non-small cell lung cancer cells via miR-338-3p/insulin receptor substrate 2. Cell Cycle. 2019; 18(24): 3525-3539.
  30. Zhang E, Shen B, Mu X, Qin Y, Zhang F, Liu Y, et al. Ubiquitin-specific protease 11 (USP11) functions as a tumor suppressor through deubiquitinating and stabilizing VGLL4 protein. Am J Cancer Res. 2016; 6(12): 2901-2909.
  31. Tajonar A, Maehr R, Hu G, Sneddon JB, Rivera-Feliciano J, Cohen DE, et al. Brief report: VGLL4 is a novel regulator of survival in human embryonic stem cells. Stem Cells. 2013; 31(12): 2833-2841.
  32. Wu A, Wu Q, Deng Y, Liu Y, Lu J, Liu L, et al. Loss of VGLL4 suppresses tumor PD-L1 expression and immune evasion. EMBO J. 2019; 38(1): e99506.
  33. Li H, Wang Z, Zhang W, Qian K, Liao G, Xu W, et al. VGLL4 inhibits EMT in part through suppressing Wnt/β-catenin signaling pathway in gastric cancer. Med Oncol. 2015; 32(3): 83.
  34. Liu X, Kong C, Zhang Z. miR-130b promotes bladder cancer cell proliferation, migration and invasion by targeting VGLL4. Oncol Rep. 2018; 39(5): 2324-2332.
  35. Deng X, Fang L. VGLL4 is a transcriptional cofactor acting as a novel tumor suppressor via interacting with TEADs. Am J Cancer Res. 2018; 8(6): 932-943.
  36. Li N, Yu N, Wang J, Xi H, Lu W, Xu H, et al. miR-222/VGLL4/ YAP-TEAD1 regulatory loop promotes proliferation and invasion of gastric cancer cells. Am J Cancer Res. 2015; 5(3): 1158-1168.
  37. Shen S, Guo X, Yan H, Lu Y, Ji X, Li L, et al. A miR-130a-YAP positive feedback loop promotes organ size and tumorigenesis. Cell Res. 2015; 25(9): 997-1012.