沉默乙酰肝素酶联合那屈肝素对肺癌细胞的抑制作用及其机制

doi:10.3969/j.issn.1004-616x.2022.06.002

Abstract

Abstract: OBJECTIVE:To investigate synergistic antitumor effects between lentivirus-mediated short hairpin RNAs (shRNA) targeting HPA and nadroparin in A549 cells,and to determine their underlying molecular mechanisms. METHODS:The lung cancer A549 cell line with down-regulated HPA expression by shRNA was used. These cells were also treated with six different conditions and then evaluated. CCK-8 assay was used to detect cell viability,enzyme-linked immunosorbent assay (ELISA) to measure concentrations of transforming growth factor (TGF-β1) in the culture medium,and the transwell assay to determine cell migration and invasion ability. Western Blot was used to detect expression of TGF-β1,phosphorylated- Smad2/3,ZEB-1,SNAIL,TWIST,E-cadherin,N-cadherin and MMP-2 in the TGF-β signaling pathway. RESULTS:HPA-silenced A549 cell line was successfully established by transfection of shRNA with lentivirus. Results from the CCK-8 assay demonstrated that shRNA inhibiting HPA or nadroparin alone slightly inhibited cell viability(all P<0.05),while the combined treatments significantly inhibited cell viability (P<0.05). ELISA results demonstrated that secretion of transforming growth factor (TGF)-β1 was decreased with inhibition of HPA and/or treatment with nadroparin (all P<0.05),particularly in the combined treatment group (P<0.05). Results from the transwell assay demonstrated that both HPA-silencing and nadroparin suppressed cell migration and invasive abilities (all P<0.05),these effects from the combined treatments were significant (all P<0.05). Western blot analyses demonstrated the similar results. Down-regulation of TGF-β1,phosphorylated-Smad2/3,Zinc-finger E-box-binding 1 (ZEB-1),twist-related protein (TWIST),snail family transcriptional repressor (SNAIL),N-cadherin and MMP-2,and upregulation of E-cadherin were induced to varying degrees by HPA-silencing or nadroparin alone or in combinations (all P<0.05). CONCLUSION:HPA silencing,especially when combined with nadroparin significantly inhibited cell invasion and migration,and up-regulated E-cadherin expression which was associated with inhibition of ZEB-1,TWIST and SNAIL,and the TGF-β1-mediated EMT process.

Key words: heparanase, nadroparin, transforming growth factor-β1, epithelial-to-mesenchymal transition, lung cancer

CLC Number:

R730.2

ZHUANG Xibing, YUAN Sujuan, ZHANG Qi, LU Minghe, CHENG Yunfeng, QIAO Tiankui. Antitumor effects of heparanase silencing plus nadroparin and the mechanisms in lung cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(6): 413-420.

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Antitumor effects of heparanase silencing plus nadroparin and the mechanisms in lung cancer cells

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