Abstract: Objective The aim of this study was to study the effect of aqueous Huaier extract on hepatocellular carcinoma and explore its potential molecular mechanism. Methods HCCLM3 cells were stimulated by different concentrations of aqueous Huaier extract as the treatment group,and untreated HCCLM3 cells as the control group.Cell viability was detected by CCK-8,cell proliferation was detected by colony formation assay,and cell metastasis was detected by Transwell assay.The expression changes of message RNA(mRNA)of HCCLM3 cells in the treatment group and control group were detected by high-throughput sequencing.The mitochondrial transcription factor A(TFAM)with obvious fold changes was selected as the research object.The results of microarray sequencing were detected by polymerase chain reaction and Western blot. Results The results of the CCK-8 experiment showed that from the third day, the Huaier 6 mg/mL group inhibited cell viability significantly stronger than the Huaier 3 mg/mL group and the control group(P＜0.05);The results showed that the number of cell clones in the Huaier 6 mg/mL group was significantly higher than that in the Huaier 3 mg/mL group and the control group(55.67±5.03 vs. 48.33±4.04 vs. 37.33±2.52,P＜0.05);Transwell test results showed that the metastatic ability of Huaier 6 mg/mL group was significantly stronger than that of Huaier 3 mg/mL group and control group(67.00±4.58 vs. 44.00±3.00 vs. 22.33±2.52,P＜0.01);high-throughput sequencing results found 486 differentially expressed genes(fold change＞2,P＜0.05),of which 317 genes were up-regulated and 115 genes were down-regulated in the experimental group.PCR experiments And Western blot experiments confirmed that Huaier can significantly reduce the expression of TFAM in liver cancer cells(1.00±0.04 vs. 0.45±0.16,P＜0.01).After Huaier stimulated HCCLM3 cells, the protein expression of p-Akt was significantly decreased, while the total AKT content did not change. Conclusion Aqueous Huaier extract can effectively inhibit the proliferation and metastasis of HCC,and this effect may be achieved by reducing the expression of TFAM and thereby inhibiting the activation of Akt signaling pathway.

Key words: Huaier, Hepatocellular carcinoma, Mitochondrial transcription factor A, Proliferation, Metastasis