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Quantitative proteomic analysis reveals the perturbation of multiple cellular pathways in jurkat-T cells induced by
Xiaoli Dong1, Lei Xiong, Xinning Jiang
1Department of Chemistry, University of California, Riverside, CA 92521-0403, USA.
Abstract:
Doxorubicin remains an important part of chemotherapy regimens in the clinic and is considered an effective agent in the treatment of acute lymphoblastic leukemia (ALL). Although the cellular responses induced by doxorubicin treatment have been investigated for years, the precise mechanisms underlying its cytotoxicity and therapeutic activity remain unclear. Here we utilized mass spectrometry, together with stable isotope labeling by amino acids in cell culture (SILAC), to analyze comparatively the protein expression in Jurkat-T cells before and after treatment with a clinically relevant concentration of doxorubicin. We were able to quantify 1066 proteins in Jurkat-T cells with both forward and reverse SILAC measurements, among which 62 were with significantly altered levels of expression induced by doxorubicin treatment. These included the up-regulation of core histones, heterogeneous nuclear ribonucleoproteins, and superoxide dismutase 2 as well as the down-regulation of hydroxymethylglutaryl-CoA synthase and farnesyl diphosphate synthase. The latter two are essential enzymes for cholesterol biosynthesis. We further demonstrated that the doxorubicin-induced growth inhibition of Jurkat-T cells could be rescued by treatment with cholesterol, supporting that doxorubicin exerts its cytotoxic effect, in part, by suppressing the expression of hydroxymethylglutaryl-CoA synthase and farnesyl diphosphate synthase, thereby inhibiting the endogenous production of cholesterol. The results from the present study provide important new knowledge for gaining insights into the molecular mechanisms of action of doxorubicin.
Insights
Doxorubicin chemotherapy for acute lymphoblastic leukemia (ALL) has unclear mechanisms. This study reveals doxorubicin inhibits cholesterol synthesis by down-regulating key enzymes, offering new insights into its cytotoxic effects.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Doxorubicin is a key chemotherapy agent for acute lymphoblastic leukemia (ALL).
- The precise mechanisms of doxorubicin's cytotoxicity and therapeutic effects remain incompletely understood.
- Understanding doxorubicin's molecular actions is crucial for optimizing cancer treatment.
Purpose of the Study:
- To investigate the protein expression changes in Jurkat-T cells following doxorubicin treatment.
- To elucidate the molecular mechanisms underlying doxorubicin's cytotoxic effects.
- To identify specific proteins and pathways affected by doxorubicin.
Main Methods:
- Utilized mass spectrometry and stable isotope labeling by amino acids in cell culture (SILAC) for comparative proteomic analysis.
- Analyzed protein expression in Jurkat-T cells before and after treatment with a clinically relevant doxorubicin concentration.
- Quantified protein expression changes to identify significantly altered proteins.
Main Results:
- Quantified 1066 proteins, with 62 showing significantly altered expression after doxorubicin treatment.
- Observed up-regulation of core histones, heterogeneous nuclear ribonucleoproteins, and superoxide dismutase 2.
- Demonstrated down-regulation of hydroxymethylglutaryl-CoA synthase and farnesyl diphosphate synthase, key enzymes in cholesterol biosynthesis.
- Showed that cholesterol supplementation could rescue doxorubicin-induced growth inhibition in Jurkat-T cells.
Conclusions:
- Doxorubicin exerts its cytotoxic effect, in part, by suppressing cholesterol biosynthesis through down-regulation of key enzymes.
- This study provides novel insights into the molecular mechanisms of doxorubicin action in cancer therapy.
- Findings contribute to a deeper understanding of doxorubicin's impact on cellular metabolism and cytotoxicity.
