Open Access Research article

Toll-like receptor 4 signaling promotes epithelial-mesenchymal transition in human hepatocellular carcinoma induced by lipopolysaccharide

Ying-Ying Jing, Zhi-Peng Han, Kai Sun, Shan-Shan Zhang, Jing Hou, Yan Liu, Rong Li, Lu Gao, Xue Zhao, Qiu-Dong Zhao, Meng-Chao Wu and Li-Xin Wei*

Author Affiliations

Tumor Immunology and Gene Therapy Center, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, PR China

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BMC Medicine 2012, 10:98  doi:10.1186/1741-7015-10-98

Published: 31 August 2012

Additional files

Additional file 1:

Supplemental figures and tables. Figure S1. MHCC97-H cells with high expression of TLR4 were induced metastasis and EMT by LPS. (A) FACS analysis for TLR4 expression in MHCC97-H cell line untreated or treated with LPS (10 μg/ml) for 48 hours. (B) The invasiveness of MHCC97-H cells pretreated with LPS (10 μg/ml) for 48 hours was determined by Transwell assay (× 200, *P < 0.05). (C) qPCR was used to detected changes in the expression of EMT genes in MHCC97-H cells treated with LPS. Results presented represent mean of triplicate experiments ± SEM (*P < 0.05). (D) Immunofluorescent staining of E-cadherin and Vimentin was performed in MHCC97-H cells that were either untreated or treated with LPS, nuclei were counterstained with DAPI (× 400). Figure S2. Upregulation of TLR4 expression in HepG2 cells and downregulation of TLR4 expreesion in SMMC-7721 cells. (A) and (B) Adeno-associated virus was used to express TLR4 in HepG2 cells. TLR4 mRNA expression was detceted by qPCR (A, *P < 0.05) and protein expression was detected by western-blot (B). (C) and (D) siRNA was used to knock down TLR4 expression in SMMC-7721 cells. TLR4 mRNA expression was detceted by qPCR (C, *P < 0.05) and protein expression was detceted by western-blot (D). Figure S3. Upregulation of Snail induced EMT in SMMC-7721 cells. Adeno-associated virus was used to express Snail in SMMC-7721 cells. E-cad mRNA expression was detected by qPCR (A*P < 0.05), and E-cad protein expression was evaluated by western-blot (B). The data shown in A and B are from one representative experiment of three performed. Figure S4. High expression of TLR4 in HCC thrombus. (A) H & E staining was performed to show thrombus in HCC tissues (× 200). (B) Immunohistochemistry was performed to show TLR4 expression in HCC thrombus as well as surrounding normal liver tissue (× 200), (a) HCC thrombus, original magnification × 400 (b) normal liver tissue, original magnification ×400. BV, blood vessels; T, thrombus; N, normal liver tissue. Table S1. Description: Sequence of the oligonucleotides for real-time PCR assays

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