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.
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
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