Abstract
BackgroundMicroRNAs(miRNAs)are involved in the initiation and progression of hepatocellular carcinoma. ESC, an extract of Stellerachamaejasme L, had been confirmed as a potential anti-tumor extract of Traditional Chinese Medicine. In light of the important role of miRNAs in hepatocellular carcinoma, we questioned whether the inhibitory effects of ESC on hepatocellular carcinoma (HCC) were associated with miRNAs.MethodsThe proliferation inhibition of ESC on HCC cells was measured with MTT assay. The migration inhibition of ESC on HCC cells was measured with transwell assay. The influences of ESC on growth and metastasis inhibition were evaluated with xenograft tumor model of HCC. Protein expressions were measured with western blot and immunofluorescence methods and miRNA profiles were detected with miRNA array. Differential miRNA and target mRNAs were verified with real-time PCR.ResultsThe results showed that ESC could inhibit proliferation and epithelial mesenchymal transition (EMT) in HCC cells in vitro and tumor growth and metastasis in xenograft models in vivo. miRNA array results showed that 69 differential miRNAs in total of 429 ones were obtained in MHCC97H cells treated by ESC. hsa-miR-107, hsa-miR-638, hsa-miR-106b-5p were selected to be validated with real-time PCR method in HepG2 and MHCC97H cells. Expressions of hsa-miR-107 and hsa-miR-638 increased obviously in HCC cells treated by ESC. Target genes of three miRNAs were also validated with real-time PCR. Interestingly, only target genes of hsa-miR-107 changed greatly. ESC downregulated the MCL1, SALL4 and BCL2 gene expressions significantly but did not influence the expression of CACNA2D1.ConclusionThe findings suggested ESC regressed growth and metastasis of human hepatocellular carcinoma via regulating microRNAs expression and their corresponding target genes.
Highlights
MicroRNAs(miRNAs)are involved in the initiation and progression of hepatocellular carcinoma
In light of the important role of miRNAs in hepatocellular carcinoma, we questioned whether the inhibitory effects of Extract of Stellerachamaejasme (ESC) on hepatocellular carcinoma were associated with miRNAs
In MHCC97H, less than inhibitory concentration of 0.05 μg/mL ESC can inhibit obviously cell migration using transwell assay and the expression of vimentin with western blot method (Fig. 2b and c). These results suggested that ESC can inhibit epithelial mesenchymal transition (EMT) in HepG2 cells and MHCC97H cells
Summary
MicroRNAs(miRNAs)are involved in the initiation and progression of hepatocellular carcinoma. ESC, an extract of Stellerachamaejasme L, had been confirmed as a potential anti-tumor extract of Traditional Chinese Medicine. Mounting evidences have demonstrated that miRNAs are involved in the initiation and. ESC, an extract of StellerachamaejasmeL, rich in isomers of Chamaejasminor, neochamaejasmine and Sikokianin [7], had antitumor effects by activating apoptosis pathway and reversed EMT of tumor cells induced by TGF-β via inhibition of Smad signaling pathway in our previous studies [8, 9]. ESC had been confirmed as a potential anti-tumor extract of Traditional Chinese Medicine. To figure out this question, this study was performed to evaluate modulatory effect of ESC on miRNAs expression in hepatocellular carcinoma in order to clarify the molecular mechanisms of ESC on hepatocellular carcinoma
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