Effect of Tetramethylpyrazine on Vascular Mimicry of A549 Caner Stem-like Cell Under Hypoxia Environment
10.13422/j.cnki.syfjx.202201721
- VernacularTitle:乏氧环境下川芎嗪对肺癌干细胞样细胞血管生成拟态的作用
- Author:
Bin LIU
1
;
Meng LI
2
;
Jie HE
1
;
Xiaoru YAN
3
;
Peitong ZHANG
1
Author Information
1. Guang' anmen Hospital， China Academy of Chinese Medical Sciences， Beijing 100053， China
2. Dongzhimen Hospital， Beijing University of Chinese Medicine， Beijing 100700， China
3. Xiyuan Hospital， China Academy of Chinese Medical Sciences， Beijing 100091， China
- Publication Type:Journal Article
- Keywords:
tetramethylpyrazine;
lung cancer;
cancer stem cells;
vascular mimicry;
hypoxia
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(23):64-70
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the effect of tetramethylpyrazine （TMP） on the vascular mimicry （VM） of non-small cell lung cancer A549 stem cell-like cells （CSLCs） in hypoxic state， and on the expression of hepatocyte growth factor （HGF）/mesenchymal-epithelial transition factor （c-Met）. MethodSerum-free sphere culture method was used to separate and enrich A549 CSLCs， and flow cytometry to detect the expression of stem cell marker CD44+/CD24-/low. CoCl2 was employed to induce hypoxia model. Cell counting kit-8 （CCK-8） assay was employed to examine the influence of 100， 200， 400， 800， 1 600， and 3 200 μmol·L-1 TMP on the viability of A549 CSLCs. The low， medium， and high concentration （100， 200， 400 μmol·L-1） of TMP that did not significantly affect the viability of A549 CSLCs was selected for subsequent experiments. Tube formation assay was used to detect the effect of different concentration of TMP on the formation of A549 CSLCs VM under hypoxia condition， and Western blot was applied to measure the expression of HGF/c-Met. ResultThe CD44+/CD24-/low expression ratio of the isolated and enriched CSLCs was （80.3±0.21）%， which was significantly higher than that of the A549 group （P<0.01）. Compared with the control group， TMP groups showed increase in the inhibition rate of CSLCs， particularly the 24 h TMP （800， 3 200 μmol·L-1） groups and the 48 h TMP （1 600， 3 200 μmol·L-1） groups （P<0.05）. Compared with blank group and Bevacizumab （Bev） group， each concentration of TMP decreased the number of tubes formed and intersections （P<0.01）. The number of tubes formed and intersections decreased in TMP groups compared with that in the SU11274 group， particularly the 200 μmol·L-1 TMP group （P<0.05） and 400 μmol·L-1 TMP group （P<0.01）. Levels of HGF and c-Met in all TMP groups were down-regulated compared with those in the blank group （P<0.05， P<0.01）. ConclusionTMP can inhibit the formation of VM in A549 CSLCs in vitro under hypoxia condition which may act by regulating HGF/c-Met related signaling pathways.
