Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2010 Dec 9;116(24):5285-8.
doi: 10.1182/blood-2010-03-272393. Epub 2010 Sep 15.

Tipifarnib sensitizes cells to proteasome inhibition by blocking degradation of bortezomib-induced aggresomes

Affiliations

Tipifarnib sensitizes cells to proteasome inhibition by blocking degradation of bortezomib-induced aggresomes

Ebenezer David et al. Blood. .

Abstract

In this report, we investigated the mechanism responsible for synergistic induction of myeloma cell apoptosis induced by the combination of tipifarnib and bortezomib. Immunofluorescence studies revealed that bortezomib alone resulted in an accumulation of puncta of ubiquitinated proteins that was further enhanced by the addition of tipifarnib. These data suggest inhibition of the degradation of bortezomib-induced aggresomes; and consistent with this possibility, we also observed an increase in p62SQSTM1 in cells treated with the combination. However, autophagy in these cells appears to be normal as LC3BII is present, and autophagic flux appears to be unaffected as demonstrated by the addition of bafilomycin A₁. Together, these data demonstrate that tipifarnib synergizes with bortezomib by inducing protein accumulation as a result of the uncoupling of the aggresome and autophagy pathways.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Growth inhibition dose-response curves. (A-D) Growth inhibition dose-response curve for MM.1S and RPMI8226 cell lines with bortezomib, tipifarnib, or the combination for 24 hours. *P < .05.
Figure 2
Figure 2
Assessment of autophagy. (A) Confocal images of RPMI8226 cell line treated with bortezomib, tipifarnib, or the combination for 24 hours. Images were acquired using a Zeiss LSM 510 confocal mounted on a Zeiss Axioplan 2 microscope. A 63× oil objective (NA 1.4) was used for image acquisition using Zeiss LSM software Version 4.2. All images were acquired with equivalent acquisition settings to allow for intensity measurements between samples. Images were processed in Adobe Photoshop (CS3) with equal contrast expansion for image display. Cells were stained with antibodies against ubiquitin and p62 SQSTM1antibody as well as 4,6-diamidino-2-phenylindole. Colocalization of ubiquitin and p62SQSTM1 in the merged image is visualized in yellow. (B) Confocal images of RPMI8226 cells that were treated with bortezomib and tipifarnib combination for 24 hours. Cells were stained for ubiquitin and then separately stained with vimentin and γ-tubulin. Colocalization of ubiquitin and vimentin in the merged image is visualized in yellow. γ-Tubulin is visualized in the periphery of ubiquitin aggregates distinct from the ubiquitin aggregates. (C) Western blotting for p62SQSTM1 and LC3BII inMM.1S cells and also in RPMI8226 cells in the presence and absence of bafilomycin A1. Relative intensity of LC3BII was calculated by normalizing the untreated control to a relative intensity of 1.0 and then dividing subsequent LC3BII band intensity by actin band intensity in the same treatment group using densitometry.

Similar articles

Cited by

References

    1. David E, Sun SY, Waller EK, et al. The combination of the farnesyl transferase inhibitor lonafarnib and the proteasome inhibitor bortezomib induces synergistic apoptosis in human myeloma cells that is associated with down-regulation of p-AKT. Blood. 2005;106(13):4322–4329. - PubMed
    1. Lonial S. Myeloma Therapy, Pursuing the Plasma Cell. Totowa, NJ: Humana Press; 2008.
    1. Lonial S, Francis D, Karanes C, et al. A phase I clinical trial testing the combination of bortezomib and tipifarnib in relapsed/refractory multiple myeloma [abstract]. Blood. 2008;112 Abstract 3706.
    1. David E, Sinha R, Chen J, et al. Perifosine synergistically enhances TRAIL-induced myeloma cell apoptosis via up-regulation of death receptors. Clin Cancer Res. 2008;14(16):5090–5098. - PubMed
    1. Kaufman J, David E, Torre C, et al. Enhanced levels of apoptosis by combination of farnesyl transferase inhibition (tipifarnib) and proteasome inhibition (bortezomib) in myeloma cell lines and primary myeloma cells and its mechanistic effects on Akt and caspase pathways. Blood. 2005;106(11):1573.

Publication types