Selinexor-Induced Tumor Regression Analysis

KT-10 tumors that regress rapidly after treatment with selinexor, were harvested 24 hours after a single dose of drug (10 mg/kg). Other, less responsive tumors, were harvested 2 hours after dose 6 (MWF dosing) at 10 mg/kg/dose. Tumors were processed for immunoblotting as previously described [30 (link)]. Immunoblots were probed for p53, p21, PARP and cleaved PARP and XPO1/CRM1. Three control and three treated tumors were analyzed for each xenograft line. GAPDH was used as a loading control. KT-10, KT-11, SK-NEP-1, CHLA258, Rh28 and Rh30 tumors were fixed in 10% buffered formalin and paraffin sections were analyzed by H&E as well with the following antibodies: FOXO1 (#2880, Cell Signaling), IKB (ab32518, Abcam), NFKB (sc-8008, Santa Cruz), pRb-phos (ab76298, Abcam), Mcl-1 (sc-819, Santa Cruz), β-catenin (#610154 BD), ERK-phos (ab32538, Abcam), survivin (ab24479, Abcam), and p53 (sc-126, Santa Cruz).

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Attiyeh E.F., Maris J.M., Lock R., Reynolds C.P., Kang M.H., Carol H., Gorlick R., Kolb E.A., Keir S.T., Wu J., Landesman Y., Shacham S., Lyalin D., Kurmasheva R.T., Houghton P.J, & Smith M.A. (2015). Pharmacodynamic and genomic markers associated with response to the XPO1/CRM1 inhibitor selinexor (KPT-330): a report from the Pediatric Preclinical Testing Program. Pediatric blood & cancer, 63(2), 276-286.

Publication 2015

ab32518 sc-8008 sc-819 ab32538 ab24479 sc-126

Antibodies Drug Formalin Gapdh Immunoblots Nfkb Paraffin Selinexor Survivin Tumors Xenograft Xpo1 β catenin

Corresponding Organization :

Other organizations : University of Pennsylvania, Children's Hospital of Philadelphia, Children's Cancer Institute Australia, Texas Tech University Health Sciences Center, Texas Tech University, Children's Hospital at Montefiore, Dupont Hospital, DuPont (United States), Duke Medical Center, St. Jude Children's Research Hospital, Karyopharm Therapeutics (United States), Nationwide Children's Hospital, Center for Cancer Research

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Variable analysis

independent variables

Selinexor treatment (10 mg/kg)
Dosing schedule (single dose vs. 6 doses)

dependent variables

Tumor regression
Protein expression (p53, p21, PARP, cleaved PARP, XPO1/CRM1)
Histological analysis (FOXO1, IκB, NF-κB, pRb-phos, Mcl-1, β-catenin, ERK-phos, survivin, p53)

control variables

Tumor type (KT-10, KT-11, SK-NEP-1, CHLA258, Rh28, Rh30)
Tumor processing (immunoblotting, paraffin sectioning)
Antibodies used for immunoblotting and immunohistochemistry
GAPDH as loading control for immunoblotting

controls

Three control tumors and three treated tumors for each xenograft line

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