Bruceantin inhibits proliferation, migration and invasion of non-small cell lung cancer H1299 cells by up-regulating miR-29a-3p
10.16438/j.0513-4870.2020-1620
- VernacularTitle:鸦胆亭上调miR-29a-3p抑制非小细胞肺癌H1299细胞增殖、迁移与侵袭
- Author:
Yan-yan YAN
1
,
2
;
Qiao-ru GUO
3
;
Xin-yue FAN
3
;
Zhuang-yan ZHU
1
;
Hai-yan ZHANG
1
;
Wen-min ZHOU
3
;
Lu-ming YANG
3
;
Dong-yu HUANG
3
;
Jian-ye ZHANG
3
Author Information
1. Institute of Immunology and School of Medicine, Shanxi Datong University, Datong 037009, China
2. Guangdong Provincial Key Laboratory of Molecular Target and Clinical Pharmacology, School of Pharmaceutical Sciences, Guangzhou Medical University, Guangzhou 511436, China
3. Guangdong Provincial Key Laboratory of Molecular Target and Clinical Pharmacology, School of Pharmaceutical Sciences, Guangzhou Medical University, Guangzhou 511436, China
- Publication Type:Research Article
- Keywords:
bruceantin;
non-small cell lung cancer;
miR-29a-3p;
integrin β1;
migration;
invasion
- From:
Acta Pharmaceutica Sinica
2021;56(2):520-527
- CountryChina
- Language:Chinese
-
Abstract:
We investigated the inhibitory effect and mechanism of action of bruceantin (BCT) on the proliferation, invasion and migration of non-small cell lung cancer (NSCLC) cells. The cytotoxic activity of BCT was measured by MTT assay; a colony forming assay, wound healing assay, and a Transwell assay were used to investigate the anti-proliferative, anti-migration, and anti-invasion effects, respectively; immunoblotting and RT-qPCR were used to detect the expression of related proteins, miRNA, and mRNA, respectively, that were involved in cell proliferation, migration, and invasion. Two gene prediction websites were used to predict the downstream target gene of miRNA. Our results show that BCT has a potent cytotoxic effect on NSCLC cell lines, with a half maximal inhibitory concentration (IC50) of BCT against H1299, PC-9, and A549 of 0.12 ± 0.02, 0.31 ± 0.20, and 2.07 ± 0.70 μmol·L-1, respectively. When H1299 cells were treated with 0.03, 0.15, and 0.75 μmol·L-1 BCT for 24 h, the proliferation, migration, and invasive ability were inhibited in a concentration-dependent manner. It is worth noting that the expression level of miRNAs related to cell migration and invasion, such as miR-29a-3p, miR-21-3p, miR-183-5p, and miR-34b-5p increased with the concentration of BCT, especially for miR-29a-3p. Using the two gene prediction websites, we predict that integrin β1 (ITGB1) may be the target gene of miR-29a-3p; immunoblot results further show that a variety of proteins related to cell proliferation, migration, and invasion, such as various proteins of the integrin family, β-catenin, p-Src, and vascular endothelial growth factor, all decreased in a concentration-dependent manner, among which the reduction of ITGB1 protein was the most obvious. RT-qPCR results showed that there was no change in ITGB1 mRNA expression. We speculate that BCT might inhibit the expression of ITGB1 protein by up-regulating miR-29a-3p independent of its mRNA level. The in-depth mechanism needs to be further explored. This study suggests that BCT has the potential for further development in the treatment of NSCLC.
