Objective To explore the effect of Hedyotisdiffusa extract(HDE)on the proliferation,migration,and invasion of liver cancer cells and the underlying mechanism.Methods Liver cancer cells HCC-7721 and LM3 induced with blank control,and low-dose and high-dose HDE for 48 h.Cell viability,proliferation and colony were assessed by cell counting kit-8(CCK-8)assay,EdU assay and colony formation,respectively.Cell migration and invasion were assessed by wound healing assay and Trasnwell assay,respectively.Cell cytoskeleton was observed by immunostaining of F-actin.RNA-seq was performed to identify differentially expressed genes in liver cancer cells induced with high-dose HDE,and verified in HCC-7721 cells by quantitative reverse-transcription polymerase chain reaction(qRT-PCR).Changes in the viability,proliferation,colony,migration and invasion of liver cancer cells induced with high-dose HDE and overexpression/knockdown of miR-340 were detected.Results HDE dose-dependently inhibited the viability,proliferation,colony,migration and invasion of HCC-7721 and LM3 cells(P＜0.05).F-actin staining found that liver cancer cells induced with blank control had more filopodia and lamellipodia,while those induced with low-dose and high-dose HDE showed shrinkage,reduced intercellular connections,and filopodia and lamellipodia.The fluorescence intensity of F-actin was significantly lower in liver cancer cells induced with low-dose and high-dose HDE than those of controls,which was significantly lower in the high-dose HDE group than the low-dose HDE group(P＜0.05).RNA-seq showed 1 769 differentially expressed genes in HCC-7721 cells induced with high-dose HDE,among which miR-340 was the mostly upregulated for 12.64 times.qRT-PCR also confirmed that HDE dose-dependently upregulated miR-340 in HCC-7721 cells(P＜0.05).High-dose HDE induction and overexpression of miR-340 significantly inhibited the viability,proliferation,colony,migration and invasion of HCC-7721 and LM3 cells(P＜0.05).Knockdown of miR-340 significantly promoted the viability,proliferation,colony,migration and invasion of HCC-7721 and LM3 cells(P＜0.05).Conclusion HDE upregulats miR-340,thereby inhibiting the cell viability,migration,and invasion abilities of liver cancer cells HCC-7721 and LM3.
Hepatocellular carcinoma/Hedyotisdiffusa/MiR-340/Neoplasm metastasis/Neoplasm invasiveness/Cell proliferation/Pharmacologic mechanisms of action(traditional Chinese medicine)
维普
万方数据