DCPIB通过上调DDIT3抑制黑色素瘤细胞增殖的机制研究

doi:10.11904/j.issn.1002-3070.2025.02.005

Abstract

Abstract: Objective The Objective of this study was to investigate the effect of DCPIB(4-[(2-butyl-6,7-dichloro-2-cyclopentyl-1-oxo-3H-inden-5-yl)oxy]butanoic acid),a selective reversible volume-regulated anion channel inhibitor,on the proliferation capacity of melanoma cells and its mechanism of action. Methods CCK-8 assay and EdU assay were used to detect the effect of DCPIB on the proliferation capacity of A375 cells and RPMI-7951 cells.Transcriptome sequencing was conducted to analyze the differentially expressed genes(DEGs)between the control group(A375 cells treated with absolute ethanol)and the DCPIB-treated group(A375 cells treated with 10 μM of DCPIB)to find the significantly changed signaling pathways.qRT-PCR was used to detect the changes in the mRNA levels of DEGs.Western blot was used to detect the changes in the expression of proteins related to relevant signaling pathways. Results DCPIB significantly inhibited the proliferation of A375 cells and RPMI-7951 cells(P＜0.01).The results of transcriptome sequencing revealed that DNA damage-inducible transcript 3(DDIT3)was significantly upregulated in the DCPIB-treated group(t=161.800,P＜0.001),and the levels of multiple key genes mRNA related to DNA damage repair pathway,including Ku70,Ku80,RAD51,MLH1,MSH6,PARP1,FANCD2 were downregulated.qRT-PCR verified that the levels of key genes mRNA in the above DNA damage repair pathways were significantly reduced after DCPIB treatment in A375 cells for 36 h(P＜0.05).GO functional enrichment and KEGG pathway analyses revealed that after DCPIB treatment,the mRNA levels of endoplasmic reticulum stress-related genes,including HSPA5,DDIT3,ATF4,XBP1,ERN1,EIF2AK3 were upregulated,and the downstream PI3K-Akt signaling pathway was abnormally expressed.qRT-PCR verified that the expression of TRIB3,a downstream molecule of DDIT3 was upregulated in A375 cells treated with DCPIB(t=2.819,P=0.048),and the downstream Akt-mTOR signaling pathway was inhibited,and the levels of p-Akt(t=7.638,P=0.002)and p-mTOR proteins(t=4.898,P=0.008)were significantly decreased. Conclusion DCPIB can significantly inhibit the proliferation of melanoma cells,and its mechanism may be related to the DDIT3-TRIB3-Akt-mTOR signaling axis.

Key words: Melanoma, A375 cells, Endoplasmic reticulum stress, DNA damage repair, Akt-mTOR signaling pathway

CLC Number:

R739.5

LI Mengya, CAI Mengdi. DCPIB inhibits melanoma cell proliferation by upregulating DDIT3[J]. Journal of Practical Oncology, 2025, 39(2): 99-107.

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URL: http://journal09.magtechjournal.com/Jwk3_syzlxzz/EN/10.11904/j.issn.1002-3070.2025.02.005

http://journal09.magtechjournal.com/Jwk3_syzlxzz/EN/Y2025/V39/I2/99

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DCPIB inhibits melanoma cell proliferation by upregulating DDIT3

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