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Metastasis is the primary cause of death of hepatocellular carcinoma (HCC), while the mechanism underlying this severe disease remains largely unclear. The Kruppel‐like factor (KLF) family is one of the largest transcription factor families that control multiple physiologic and pathologic processes by governing the cellular transcriptome. To identify metastatic regulators of HCC, we conducted gene expression profiling on the MHCC97 cell series, a set of subclones of the original MHCC97 that was established by in vivo metastasis selection therefore harbouring differential metastatic capacities. We found that the expression of KLF9, a member of the KLF family, was dramatically repressed in the metastatic progeny clone of the MHCC97 cells. Functional studies revealed overexpression of KLF9 suppressed HCC migration in vitro and metastasis in vivo, while knockdown of KLF9 was sufficient to promote cell migration and metastasis accordingly. Mechanistically, we found the expression of KLF9 can reverse the pro‐metastatic epithelial‐mesenchymal transition (EMT) program via direct binding to the promoter regions of essential mesenchymal genes, thus repressing their expression. Interestingly, we further revealed that KLF9 was, in turn, directly suppressed by a mesenchymal transcription factor Slug, suggesting an intriguing negative feedback loop between KLF9 and the EMT program. Using clinical samples, we found that KLF9 was not only downregulated in HCC tissue compared to its normal counterparts but also further reduced in the HCC samples of whom had developed metastatic lesions. Together, we established a critical transcription factor that represses HCC metastasis, which is clinically and mechanically significant in HCC therapies. [ABSTRACT FROM AUTHOR]

ABSTRACT Introduction The large-leaved Kudingcha from the genus Ilex, which is used as a traditional Chinese tea, contains several characteristic triterpenoid saponins that can be subjected to quality control evaluation. Objective To develop and validate a rapid method incorporating reverse-phase ultra-performance liquid chromatography coupled with evaporative light scattering detection (UPLC-ELSD) for the simultaneous determination of the five triterpenoid saponins kudinoside L (1), kudinoside C (2), kudinoside A (3), kudinoside F (4) and kudinoside D (5) in several species of the large-leaved Kudingcha from the genus Ilex and 'Yerba Mate' ( Ilex paraguariensis). Methodology The five compounds were separated using a water-acetonitrile mobile phase with a Waters Acquity BEH C18-column (100 × 2.1 mm, 1.7 µm). Results Separation took 13 min with detection and quantification limits ranging from 12.5 to 29.8 ng and 41.3 to 98.2 ng, respectively. The method was validated according to the regulatory guidelines with respect to precision, stability, repeatability and recovery. The triterpenoid saponins showed a good regression relationship ( r2 > 0.999) within the test ranges, and the recovery of the method was in the 95-105% range. Conclusion The present method can be used successfully for the quality control of the large-leaved Kudingcha. The different Ilex species showed differences in distribution of the five triterpenoids. Ilex kudingcha, which makes up the major species of the large-leaved Kudingcha, contains the maximum amount of triterpenoid saponins. Copyright © 2012 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]