Study on the Mechanism of Cucurbitacin B Inducing Apoptosis in HuCCT1 cells of Intrahepatic Cholangiocarcinoma Based on Metabonomics
Objective To explore the role and mechanism of cucurbitacin B(CuB)in inducing apoptosis of human intrahepatic cholangiocarcinoma HuCCT1 cells.Methods HuCCT1 cells were divided into control group,low,medium and high dose groups(50,100 and 200 nmol·L-1)of CuB,and drug intervention was given for 24 hours.Annexin V/PI double staining and Hoechst staining were used to detect apoptosis.The main differential metabolites were detected by ultrahigh performance liquid chromatography/quadrupole time of flight mass spectrometry(UHPLC-Q/TOF-MS),and the related metabolic pathways were analyzed.Results The results of Annexin V/PI double staining showed that compared with the control group,the apoptotic ratio of HuCCT1 cells treated with CuB increased from 7.86%±0.73%to 16.53%±0.27%(P<0.0001);Hoechst staining results showed that compared with the control group,after 24 h of treatment with 200 nmol·L-1 CuB,the cell structure was destroyed,the cells were broken,the edge contour was irregular,and a lot of karyopyknosis appeared;20 significantly different metabolites were screened by metabonomic analysis,mainly involving TCA cycle,energy metabolism,amino acid metabolism and lipid metabolism.Conclusion CuB has a good anti-tumor effect,which may play a pharmacological role by interfering with ultiple metabolic pathways of HuCCT1 cells,causing a lack of substances for rapid cell proliferation,and ultimately leading to apoptosis.
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