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Beta-Hydroxyisovalerylshikonin Inhibits the Growth of U266 Multiple Myeloma Cells by Triggering the Mitochondrial Pathway

Yihui He, Xiaoyan Chen and Lingying Kong

Background: Beta-hydroxyisovalerylshikonin (beta-HIVS) is a compound isolated from the traditional oriental medicinal herb lithospermum radix. This drug exerts a role as an ATP non-competitive inhibitor of Protein-Tyrosine Kinases (PTKs) and shows great potential for induction of apoptosis against human cancer cells. We investigated the effect of beta-HIVS on multiple myeloma U266 cells and clarified details of the primary mechanism in its apoptosisinducing activity. Objective: The aim of this work was to trace the apoptosis-inducing activity of beta-HIVS on U266 cells as well as the underlying mechanisms. Methods: Cell Counting Kit-8 (CCK-8) test and colony-forming assay were performed in estimating the effects of beta-HIVS on U266 cell viability and colony formation. Apoptosis analysis was carried out on the basis of DAPI fluorescence staining and DNA fragmentation assays. Real-time PCR was employed to evaluate changes of Bcl-2 and Bax mRNA expression, while indirect immunofluorescence assay and western blotting were utilized in validating the expression of Bcl-2, Bax, caspase-3, caspase-9, PARP and cytochrome c. Results: CCK-8 test and colony-forming assay showed that beta-HIVS treatment resulted in significantly reduced cell proliferation (P