Experimental Study of Larotrectinib Regulating Autophagy and Inhibiting Proliferation and Migration of Colon Cancer Cells Through AMPK/mTOR Signaling Pathway
10.3969/j.issn.1671-7414.2024.06.005
- VernacularTitle:拉罗替尼通过AMPK/mTOR信号通路调控结肠癌细胞自噬和抑制增殖与迁移的实验研究
- Author:
Xihui BAI
1
;
Shiyu LIU
;
Yuanyuan SUN
Author Information
1. 西安交通大学第一附属医院药学部,西安 710061
- Keywords:
colon cancer;
larotrectinib;
AMPK/mTOR pathway;
autophagy
- From:
Journal of Modern Laboratory Medicine
2024;39(6):29-36
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of Larotrectinib(Lar)on autophagy,proliferation and migration of colon cancer cells and its molecular mechanism.Methods Human colon cancer cell lines COLO 205,HCT 116 and human colonic epithelial cell line CP-H040 were treated with different concentrations of Lar(0,100,200,400,800,1600 and 3 200 nmol/L).CCK-8 assay was used to detect the cell viability of COLO 205,HCT 116 and CP-H040 cells.COLO 205 and HCT 116 cells were randomly divided into control group(Con group),Lar group,Chloroquine group(CQ group)and Lar+CQ group.Cell invasion was detected by Transwell assay.Scratch test was used to detect cell migration ability.Ki67 immunofluorescence staining was used to detect cell proliferation.Real-time quantitative polymerase chain reaction was used to detect the mRNA expression of epithelial-mesenchymal transition related markers in colon cancer cells.Autophagy was detected by adenovirus transfection experiment and transmission electron microscopy.Western blot was used to detect the expression of adenosine monophosphate-activated protein kinase/mammalian target of rapamycin(AMPK/mTOR)pathway related proteins.Results Lar significantly inhibited the viability of COLO 205 and HCT 116 cells in a concentration-dependent manner(F=355.181,403.758,all P<0.001).Compared with the Con group,the number of invasive cells,Ki67 fluorescence intensity and scratch healing rate of colon cancer cells in the Lar group were decreased,the expression of E-cadherin mRNA was increased,the expressions of Vimentin and MMP2 mRNA were decreased,the formation of autophagosomes and autophagic flow,the ratio of microtubule-associated protein light chain 3(LC3)Ⅱ/Ⅰ and p-AMPK/AMPK were increased,and the expression of p62 protein and p-mtor/mTOR ratio were decreased,with significant differences(t=4.399～54.214,all P<0.05).Compared with the Lar group,the formation of autophagosome was decreased and the expression of p62 protein was increased in the Lar+CQ group,and the difference was statistically significant(t=2.755～24.784,all P<0.05).Conclusion Lar can inhibit the proliferation and migration of colon cancer cells,and the underlying mechanism is related to activation of the AMPK/mTOR signaling pathway and thus inducts autophagy.
