| ?Background?Hepatocellular carcinoma(HCC)is the most common primary liver cancer.It accounts for about 70% of the liver cancer cases.And it is one of the cancers that seriously endanger the health of the human body.There are many treatment methods for liver cancer,such as surgical resection,liver transplantation,transcatheter arterial chemoembolization,and systemic chemotherapy.Surgical resection and liver transplantation are potential treatments.Most patients are diagnosed with advanced cancer,so only a few patients can be treated with these types of treatment.Systematic treatment is common in clinic.However,patients who with advanced hepatocellular carcinoma usually have other liver diseases.The patients have poor systemic chemotherapy tolerance.And chemotherapy has great toxic and side effects,which makes the treatment effect unsatisfactory.Chinese medicine has a long history.At present,traditional Chinese medicine has been widely used to treat cancer.Cinobufotalin is a common Chinese medicine,which is extracted from the secretions of toads.The main components of Cinobufotalin are bufadienolides,alkaloids,peptides and cholesterol.The functions of Cinobufotalin are clearing away heat and detoxification,reducing inflammation and analgesic,and enhancing immunity.Many studies have shown that Cinobufotalin has antitumor activity and enhances the effect of chemotherapy,and has the advantages of low toxicity.And it using safely and has no addiction.Cinobufotalin has antitumor effects on various tumors such as non-small cell lung cancer,colorectal cancer,liver cancer,gastric cancer and esophageal cancer.There are various dosage forms of Cinobufotalin preparations in clinical using,such as Cinobufotalin injection,Cinobufotalin capsules,and Cinobufotalin oral solution.In this paper,we will study the effect of Cinobufotalin on hepatocellular carcinoma cells.And we will study the mechanism of hepatocellular carcinoma cell apoptosis induced by Cinobufotalin.The results can provide a more effective theoretical basis for the clinical application of Cinobufotalin.?Objective?In this study,hepatocellular carcinoma cells were taken as the research object.And we studied the effect of Cinobufotalin on the cell viability of liver cancer cells and verified it through in vivo experiments.Next,we studied the effect of Cinobufotalin on the mitochondrial function of hepatoma cells,and we initially explored the mechanism of Cinobufotalin-induced apoptosis in hepatoma cells.?Methods?1.By using MTT,LDH,wound healing test,EDU test,clonogenic experiment,DAPI,flow cytometry,TUNEL to detect the effect of Cinobufotalin on hepatoma cell viability,proliferation ability and cell apoptosis.2.Tumor xenotransplantation and immunohistochemical experiments were conducted to verify the inhibitory effect of Cinobufotalin on hepatocellular carcinoma cells in vivo.3.To study the effect of Cinobufotalin on mitochondrial function of liver cancer cells by using mitochondrial membrane potential,ATP,Western blot and RT-q PCR methods.?Results?1.The increased concentration of Cinobufotalin can significantly inhibit the proliferation and cell activity of liver cancer cells and induce cell apoptosis.The results of MTT,microscopic observation and LDH experiment showed that Cinobufotalin has an inhibitory effect on the viability of liver cancer cells.The results of clonogenic experiment and EDU experiment showed that Cinobufotalin can inhibit the proliferation of liver cancer cells.The results of the cell scratch test showed that Cinobufotalin has an inhibitory effect on the migration ability of liver cancer cells;the results of DAPI staining experiment,flow cytometry and TUNEL test showed that Cinobufotalin can promote the apoptosis of liver cancer cells.2.The results of tumor xenotransplantation and immunohistochemical experiments showed that Cinobufotalin can significantly inhibit tumor formation and proliferation in nude mouse.3.Cinobufotalin can significantly increase mitochondrial membrane potential and inhibit the production of ATP in liver cancer cells.The expression of apoptosis-related molecule Bax was up-regulated,the expression of bcl-2 was down-regulated,and the phosphorylation level of p65 was significantly reduced,while the total level of p65 showed no significant change.?Conclusion?Our results showed that Cinobufotalin could inhibit the proliferation of hepatocellular carcinoma cells SNU-739 and induce the apoptosis of hepatocellular carcinoma cells through the mitochondrial pathway. |
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