Objective: To investigate the inhibitory effect of Elesclomol (ES) + Cu2 + on the proliferation of human hepatoma cell lines PLC/PRF/5 and Huh-7 and its potential to induce Cuproptosis. Methods: Human hepatoma cell lines PLC/PRF/5 and Huh_7 cells were Cultured in vitro. ES solution , Cu2 + solution and copper chelating agent ammonium tetrathiomolybdate VI (ATTM) solution was treated separately or in combination. The effect of ES + Cu2 + on the survival rate of human hepatoma cell lines PLC/PRF/5 and Huh_7 cells and the effect of ES + Cu2 + on the survival rate after pretreatment with copper chelating agent ATTM were evaluated using CCK_8 kit. The cell death induced by ES + Cu2 + was detected by flow cytometry and the changes of ES + Cu2 + after pretreatment with copper chelating agent ATTM. The expression of Cuproptosis related proteins ATPase copper transporting beta (ATP7B) ,ferredoxin 1 (FDX1) , dihydrolipoamide s_acetyltransferase(DLAT) and superoxide dismutase 1 (SOD1) were detected by Western blot. The effect of ES + Cu2 + on cell proliferation and the reverse effect after ATTM pretreatment was detected by cell scratch assay. Results: The toxicity of ES + Cu2 + to human hepatocellular carcinoma cell lines PLC/PRF/5 and Huh_7 was significantly dose_dependent (P < 0. 05) . Compared with the control group , the combined application of ES and Cu2 + had a more significant inhibitory effect on hepatocellular carcinoma cells than ES or Cu2 + alone (P < 0. 05) , and copper chelating agent ATTM could reverse the inhibitory effect of ES + Cu2 + on hepatocellular carcinoma cells (P < 0. 05) . Flow cytometry results showed that compared with the control group , the proportion of cell death in PLC/PRF/5 and Huh_7 cells treated with ES + Cu2 + increased , while the proportion of cell death decreased after ATTM intervention (P < 0. 05) . The results of cell scratch test showed that the migration ability of PLC/PRF/5 and Huh_7 cells was decreased after ES + Cu2 + treatment , however, the addition of ATTM reversed the inhibitory effect of ES + Cu2 + on cell migration (P < 0. 05) . Compared with the control group , the expression levels of copper death related proteins ATP7B , FDX1 , DLAT and SOD1 decreased after ES + Cu2 + treatment , but the addition of ATTM reversed the expression trend of these proteins (P < 0. 05) . Conclusion The combination of ES and Cu2 + can effectively inhibit the proliferation and migration of PLC/PRF/5 and Huh_7 of hepatocellular carcinoma cells , and induce Cuproptosis , which provides a new strategy for the treatment of hepatocellular carcinoma.

Objective: To investigate the effects of Krüppel-like factor 4 (KLF 4) knockdown on epithelialmesenchymal transition (EMT), cell migration and invasion of prostate cancer cells. Methods: The prostate cancer cells with stable knockdown of KLF 4 named as LNCaP-shKLF4 cells and the LNCaP-con cells as the control were constructed. The expressions of KLF4 mRNA and protein in LNCaP-con and LNCaP-shKLF4 cells, and the expressions of EMTassociated gene mRNA and protein in the prostate cancer PC3-shKLF4 cells with stable knockdown of KLF 4 and the PC3-con cells (as the control) as well as the LNCaP-shKLF4 and LNCaP-con cells were detected by real-time fluorescence quantitative-PCR and Western blotting, respectively. The abilities of migration and invasion of PC3-shKLF4 and PC3-con cells as well as LNCaP-shKLF4 and LNCaP-con cells were detected by Transwell chamber assay. Results: LNCaP-shKLF4 and LNCaP-con cells were successfully constructed. The expression levels of KLF4 mRNA and proteins in LNCaP-shKLF4 cells were lower than those in LNCaPcon cells (P<0.01, P<0.05). The expression levels of E-cadherin (E-cad) mRNA in PC3- shKLF4 and LNCaP-shKLF4 cells were higher than those in PC3-con and LNCaP-con cells, respectively (both P<0.01). The expression levels of N-cadherin (N-cad), Zinc finger E-box-binding homeobox 1 (Zeb1), Snail1, vimentin (Vim) and matrix metallopeptidase 1 (MMP1) mRNAs in PC3-shKLF4 and LNCaP-shKLF4 cells were lower than those in PC3-con and LNCaP-con cells, respectively (all P<0.05). The expression levels of E-cad protein in PC3-shKLF4 and LNCaP-shKLF4 cells were higher than those in PC3-con and LNCaP-con cells, respectively (both P<0.05). The expression levels of N-cad, Zeb1, Snail1, Vim and MMP1 mRNAs in PC3-shKLF4 and LNCaP-shKLF4 cells were lower than those in PC3-con and LNCaP-con cells, respectively (all P<0.05). The abilities of migration and invasion of PC3-shKLF4 and LNCaP-shKLF4 cells were weaker than those of PC3-con and LNCaP-con cells, respectively (P<0.01, P<0.05). Conclusion: KLF 4 knockdown in prostate cancer cells can activate the expression of epithelium-associated genes and inhibit the expressions of mesenchymal-associated genes, resulting in the inhibition of cell migration and invasion of prostate cancer cells in vitro.