Mechanism of the inhibitory effects of thalidomide on expressions of VEGF and bFGF

doi:10.12047/j.cjap.6262.2022.032

Abstract

Abstract: Objective: To investigate the inhibitory effects of thalidomide on the expressions of VEGF and bFGF in human lung adenocarcinoma A549 cells and human hepatocellular carcinomas HepG2 cells mediated by cereblon (CRBN). Methods: shRNA technology was used to construct the A549 cell line (A549_CRBN) and HepG2 cell line (HepG2_CRBN) with stable knockdown of CRBN, which was verified by real-time PCR and Western blot. A549 cells were divided into negative control group (A549_luciferase) and CRBN down-regulation group (A549_CRBN); HepG2 cells were divided into negative control group (HepG2_luciferase) and CRBN down-regulation group (HepG2_CRBN). The above cells were seeded into 6-well plates at 3×10⁵ cells/well, and cultured in a 37℃, 5% CO₂ incubator for 24 h. Then, 1 ml medium containing 100 μmol/L thalidomide (thalidomide group) and 1 ml medium containing 1‰ DMSO (control group) were added respectively, and the culture was continued for 24 hours before subsequent experiments. Each group was designed with three replicate wells. The effect of thalidomide on the activity of A549 cell line was detected by MTS assay. Real-time PCR was performed to detect mRNA expression levels of VEGF, bFGF and c-jun. ELISA assay was performed to detect protein expressions of VEGF and bFGF. Results: Compared with the control group, thalidomide at the concentrations of 1, 10, 50 and 100 μmol/L had no significant effects on the proliferation of A549 and HepG2 cells (P＞0.05). VEGF and bFGF levels in the A549_CRBN or HepG2_luciferase groups were significantly lower than those in the A549_CRBN or HepG2_CRBN groups (P＜0.05). Compared with the control group of the A549_luciferase or HepG2_luciferase, thalidomide inhibited the expressions of VEGF and bFGF in A549_luciferaseand HepG2_luciferase cells (P＜ 0.05), but did not inhibit the expressions of VEGF and bFGF in A549_CRBN and HepG2_CRBN cells. Compared with the control group of the HepG2_luciferase, thalidomide inhibited c-Jun expression in HepG2_luciferase cells (P＜0.01), but did not significantly inhibit c-Jun expression in HepG2_CRBN cells. Conclusion: The inhibitory effects of thalidomide on VEGF and bFGF expressions may be mediated by CRBN in A549 and HepG2 cells, and c-Jun may be one of the key transcription factors responsible for this inhibition.

Key words: thalidomide, angiogenesis, cell culture, VEGF, bFGF, cereblon(CRBN)

CLC Number:

R966

HUANG Hai-ning, WANG Hui, LONG Chao-liang, ZHANG Hao, WANG Hai. Mechanism of the inhibitory effects of thalidomide on expressions of VEGF and bFGF[J]. CJAP, 2022, 38(2): 169-174.

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