|Table of Contents|

Effects of targeted silencing fatty acid synthase FASN gene on apoptosis and metastasis level of esophageal cancer cells in lost nests

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2023 15
Page:
2809-2815
Research Field:
Publishing date:

Info

Title:
Effects of targeted silencing fatty acid synthase FASN gene on apoptosis and metastasis level of esophageal cancer cells in lost nests
Author(s):
YU Fangfang1; LI Juyuan1; HUANG Jian1; LYU Qiongxiu2
1.Laboratory Department;2.Department of Oncology,Hainan Western Central Hospital,Hainan Danzhou 571700，China.
Keywords:
esophageal carcinoma; fatty acid synthase; anoikis apoptosis; epithelial mesenchymal transformation; migration; invasion
PACS:
R735.1
DOI:
10.3969/j.issn.1672-4992.2023.15.009
Abstract:
Objective:To investigate the effects of silencing fatty acid synthase(FASN) on apoptosis and metastasis of esophageal cancer cells.Methods:Real-time quantitative fluorescent PCR(qRT-PCR) was used to detect FASN mRNA expression levels in human normal esophageal epithelial cell lines(HEEC) and human esophageal cancer cell lines(CaES-17,EC109,EC9706,KYSE170,TE-1).The experiment was divided into Control group,NC-shRNA group,FASN-SHRNA1 group and FASN-SHRNA2 group.Lentivirus-mediated NC-shRNA,FASN-shRNA1 group and FASN-shRNA2 group were transfected into EC109 cells,respectively.The inverted microscope was used to observe the green fluorescense.qRT-PCR and Western blot were used to detect the transfection effect.The apoptosis of EC109 cells in lost nests was simulated by polyhydroxy-hema(Poly-HEMA) coated culture plate.AnnexinV-FITC/PI double staining was used to detect the apoptosis.Calcein AM/EthD-1 fluorescence double staining was used to identify the living and dead cells.Fluorescence staining of E-cadherin and Vimentin in EC109 cells of each group was observed by immunofluorescence staining.The protein expressions of E-cadherin,Vimentin,Snail,matrix metalloproteinase 2(MMP-2) and MMP-9 in EC109 cells of each group were determined by Western blot.Transwell assay was used to detect the migration and invasion of EC109 cells.Results:Compared with HEEC,the relative expression level of FASN mRNA in human esophageal cancer cell lines CaES-17,EC109,EC9706,KYSE170,TE-1 was increased(P＜0.05).The green fluorescence expression was obvious in the NC-shRNA group,the FASN-SHRNA1 group and the FASN-SHRNA2 group,compared with the NC-shRNA group,the relative mRNA expression and protein expression of FASN in the FSN-SHRNA1 group and the FASN-SHRNA2 group were increased(P＜0.05),the cell apoptosis rate was increased(P＜0.05),the positive rate of EthD-1 labeled cells was also increased(P＜0.05),the intensity of intracellular E-cadherin fluorescence staining was increased,the intensity of Vimentin fluorescence staining was decreased,the relative expression of E-cadherin protein was up-regulated(P＜0.05),and the protein relative expression levels of Vimentin,Snail,MMP-2 and MMP-9 were down-regulated(P＜0.05),and the number of cell migration and invasion were decreased(P＜0.05).Conclusion:Lentivirus-mediated FGN-shRNA silencing FASN expression can promote the apoptosis of esophageal cancer cells and inhibit the migration and invasion induced by epithelial mesenchymal transformation.

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Memo

Memo:
海南省卫生健康行业科研项目（编号：22A200041）
Last Update: 2023-06-30