Study on the molecular mechanism of cat's claw ethanol extract in regulating apoptosis of colorectal cancer cell HCT116
Objective To explore the potential molecular mechanism of cat's claw ethanol extract(CCEE)regulating the apoptosis of colorectal cancer cell HCT116.Methods The ethanol extract of cat's claw was used to treat colorectal cancer cells HCT116,and the apoptosis level was detected.Colorectal cancer cells HCT116 were analyzed by semi-quantitative mass spectrometry after treatment with ethanol extract of cat's claw.After overexpressing the differential protein,the apoptosis level of colorectal cancer cell HCT116 was detected.miRNAs of differential proteins were detected through miRDB online analysis and luciferase reporter system.Results After CCEE treatment,the apoptosis level of colorectal cancer cell HCT116 was increased[(23.18±3.15)％vs.(68.21±12.05)％,t=6.26,P＜0.05].After 24 h treatment with CCEE,the expression level of BAX protein in colorectal cancer cell HCT116 was increased by semi-quantitative proteomics(P＜0.05).When BAX was overexpressed,the apoptosis level of colorectal cancer cell HCT116 was increased[(28.00±6.68)％vs.(85.08±4.97)％,t=1 1.87,P＜0.05].The expression level of miR-761 was significantly decreased after CCEE treatment of colorectal cancer cells HCT116[(0.82±0.10)vs.(0.11±0.02),t=12.00,P＜0.05].After overexpression of miR-761,the protein level and mRNA level of BAX were decreased[(0.061±0.003)vs.(0.004±0.001),t=16.08,P＜0.05].The apoptosis level was decreased[(21.59±5.66)％vs.(6.04±2.13)％,t=4.45,P＜0.05].In addition,miR-761 targeted the 3-terminal non-coding region of BAX mRNA[(100.00±9.59)vs.(14.22±2.45),t=15.01,P＜0.05].Conclusion Cat's claw ethanol extract could inhibit the expression level of miR-761 in colorectal cancer cells HCT116,reduce the degradation of BAX mRNA due to miR-761,enhance the protein translation of BAX,and promote colorectal cancer cells HCT116 apoptosis.
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