Timosaponin A-Ⅲ suppressing NSCLC growth through regulation of the ASAP1-IT1/DNMT3b/YAP1 axis

  • FU Xiangjun ,
  • WANG Mei ,
  • ZHANG Yuan ,
  • LI Yanna ,
  • GENG Biao ,
  • ZHAO Wenying
Expand
  • 1. Wannan Medical College, Wuhu 241002, China;
    2. The First Affiliated Hospital of Wannan Medical College

Received date: 2022-07-24

  Online published: 2023-01-09

Abstract

Objective: To explore the effect and mechanism of Timosaponin A-Ⅲ (TAⅢ) on NSCLC growth. Methods: A549 cells was processed with different concentrations of TAIII (5, 10, 15, 20, 25, 30 μmol/L), the cell viability was measured by CCK-8. qRT-PCR was used to detect the expressions of Adenosine diphosphate ribosylation factor guanylate kinase 1-intronic transcript 1(ASAP1-IT1) and Yes-associated Protein 1(YAP1) in tissues and cells. EDU, flow cytometry and Western blot were used to detect ASAP1-IT1, DNA (cytosine-5-)-methyltransferase 3β (DNMT3b), YAP1 and their effects on cell proliferation, apoptosis rate and the influence of apoptosis related genes (Bax and caspase-3) and anti-apoptotic gene Bcl2. Bisulfite sequencing PCR (BSP) was used to analysis the relationship between ASAP1-IT1 and YAP1. RNA immunoprecitation (RIP) and qRT-PCR assays were used to analyze the interaction between ASAP1-IT1 and DNMT3b. Co-immunoprecipitation (Co-IP) was used to confirm the interaction between YAP1 and DNMT3b. qRT-PCR method was used to analyze the effect of TAⅢ on the expression of ASAP1-IT1/DNMT3b/YAP1 axis-related genes. Results: ASAP1-IT1 and YAP1 was up-regulated in NSCLC tissues and cancer cells, and DNMT3b was down-regulated in NSCLC tissues and cancer cells, the levels of ASAP1-IT1, YAP1 and DNMT3b were reversed after using TAⅢ. Silencing of ASAP1-IT1 and YAP1, overexpression of DNMT3b or taking TAⅢ significantly increased apoptosis rate of A549 cells and apoptosis related indexes. TAⅢ reversed the silencing or overexpression of ASAP1-IT1, DNMT3b and YAP1. Furthermore, ASAP1-IT1 was found to interact with DNMT3b, and DNMT3b and YAP1 were interacted. Conclusion: ASAP1-IT1/DNMT3b/YAP1 axis promotes NSCLC progression by regulating cell proliferation and apoptosis. TAⅢ suppresses proliferation and improve apoptosis of lung cancer cells by regulating ASAP1-IT1/DNMT3b/ YAP1 axis.

Cite this article

FU Xiangjun , WANG Mei , ZHANG Yuan , LI Yanna , GENG Biao , ZHAO Wenying . Timosaponin A-Ⅲ suppressing NSCLC growth through regulation of the ASAP1-IT1/DNMT3b/YAP1 axis[J]. Journal of Baotou Medical College, 2022 , 38(11) : 10 -17 . DOI: 10.16833/j.cnki.jbmc.2022.11.003

References

[1] Russell é, Conroy MJ, Barr MP. Harnessing natural killer cells in non-small cell lung cancer[J]. Cells, 2022, 11(4): 605.
[2] Mithoowani H, Febbraro M. Non-small-cell lung cancer in 2022: a review for general practitioners in oncology[J]. Curr Oncol, 2022, 29(3): 1828-1839.
[3] Lin Y, Zhao WR, Shi WT, et al. Pharmacological activity, pharmacokinetics, and toxicity of timosaponin AIII, a natural product isolated from Anemarrhena asphodeloides bunge: a review[J]. Front Pharmacol, 2020, 11: 764.
[4] Liu JJ, Deng XH, Sun XJ, et al. Inhibition of autophagy enhances timosaponin AIII-induced lung cancer cell apoptosis and anti-tumor effect in vitro and in vivo[J]. Life Sci, 2020, 257: 118040.
[5] Jung O, Lee J, Lee YJ, et al. Timosaponin AIII inhibits migration and invasion of A549 human non-small-cell lung cancer cells via attenuations of MMP-2 and MMP-9 by inhibitions of ERK1/2, Src/FAK and β-catenin signaling pathways[J]. Bioorg Med Chem Lett, 2016, 26(16): 3963-3967.
[6] Cheng ZA, Lu CL, Wang H, et al. Long noncoding RNA LHFPL3-AS2 suppresses metastasis of non-small cell lung cancer by interacting with SFPQ to regulate TXNIP expression[J]. Cancer Lett, 2022, 531: 1-13.
[7] Kwaŝniak K, Czarnik-kwaŝniak J, Malysheva K, et al. LncCDH5-3: 3 regulates apoptosis, proliferation, and aggressiveness in human lung cancer cells[J]. Cells, 2022, 11(3): 378.
[8] Cao Z, Oyang L, Luo X, et al. The roles of long non-coding RNAs in lung cancer[J]. J Cancer, 2022, 13(1): 174-183.
[9] Zhang L, Shi SB, Zhu Y, et al. Long non-coding RNA ASAP1-IT1 promotes cell proliferation, invasion and metastasis through the PTEN/AKT signaling axis in non-small cell lung cancer[J]. Eur Rev Med Pharmacol Sci, 2018, 22(1): 142-149.
[10] Liu YT, Yang YP, Zhang LL, et al. LncRNA ASAP1-IT1 enhances cancer cell stemness via regulating miR-509-3p/YAP1 axis in NSCLC[J]. Cancer Cell Int, 2021, 21(1): 572.
[11] Yang Q, Yang D, Zhu HB, et al. LncRNA MORT negatively regulates FGF1 to suppress malignant progression of breast cancer[J]. Eur Rev Med Pharmacol Sci, 2021, 25(20): 6179-6186.
[12] 刘艳平. 知母皂苷成分的药理活性及作用机制研究进展[J]. 药学实践杂志, 2018, 36(1): 24-29.
[13] Wang K, Hu YB, Zhao Y, et al. Long non-coding RNA ASAP1-IT1 suppresses ovarian cancer progression by regulating Hippo/YAP signaling[J]. Int J Mol Med, 2021, 47(4): 44.
Outlines

/