Sulfurtransferases and Cystathionine Beta-Synthase Expression in Different Human Leukemia Cell Lines

Abstract

The studies concerned the expression of sulfurtransferases and cystathionine beta-synthase in six human leukemia cell lines: B cell acute lymphoblastic leukemia-B-ALL (REH cells), T cell acute lymphoblastic leukemia-T-ALL (DND-41 and MOLT-4 cells), acute myeloid leukemia-AML (MV4-11 and MOLM-14 cells), and chronic myeloid leukemia-CML (K562 cells). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis were performed to determine the expression of thiosulfate sulfurtransferase, 3-mercaptopyruvate sulfurtransferase, gamma-cystathionase, and cystathionine beta-synthase on the mRNA and protein level. Interestingly, we found significant differences in the mRNA and protein levels of sulfurtransferases and cystathionine beta-synthase in the studied leukemia cells. The obtained results may contribute to elucidating the significance of the differences between the studied cells in the field of sulfur compound metabolism and finding new promising ways to inhibit the proliferation of various types of leukemic cells by modulating the activity of sulfurtransferases, cystathionine beta-synthase, and, consequently, the change of intracellular level of sulfane sulfur as well as H₂S and reactive oxygen species production.

Keywords: 3-mercaptopyruvate sulfurtransferase; cystathionine beta-synthase; gamma-cystathionase; l-cysteine; leukemia cells; thiosulfate sulfurtransferase.

Figure 1

The different types of human leukemia cell lines. (Created with BioRender.com, accessed on 14 January 2022).

Figure 2

The expression of TST, MPST, CBS, and CTH on the mRNA level in leukemia cell lines (RT-PCR analysis). β-Actin was used as the loading control. Data from a representative experiment are presented. All the experiments were repeated at least three times with similar results.

Figure 3

The expression of TST, MPST, CBS, and CTH on the protein level in leukemia cell lines (Western blot analysis). β-Actin was used as the loading control. Data from a representative experiment are presented. All the experiments were repeated at least three times with similar results.

Figure 4

The expression on the mRNA (A) and protein (B) level of thiosulfate sulfurtransferase in leukemia cell lines (REH, DND-41, MOLT-4, MV4-11, MOLM-14, K562). β-Actin was used as a loading control. Quantification of the TST expression on mRNA level (RT-PCR method) and protein level (Western blotting method) was performed by densitometric analysis of the gels/blots and normalized to the internal loading control. The results are expressed as the mean ± SD of three or more independent experiments. Each mark represents p < 0.05 as followed: * versus REH; # versus DND-41; $ versus MOLT-4.

Figure 5

The expression on the mRNA (A) and protein (B) level of 3-mercaptopyruvate sulfurtransferase in leukemia cell lines (REH, DND-41, MOLT-4, MV4-11, MOLM-14, K562). β-Actin was used as a loading control. Quantification of the MPST expression on mRNA level (RT-PCR method) and protein level (Western blotting method) was performed by densitometric analysis of the gels/blots and normalized to the internal loading control. The results are expressed as the mean ± SD of three or more independent experiments. Each mark represents p < 0.05 as followed: * versus REH; # versus DND-41; $ versus MOLT-4.

Figure 6

The expression on the mRNA (A) and protein (B) level of cystathionine β-synthase in leukemia cell lines (REH, DND-41, MOLT-4, MV4-11, MOLM-14, K562). β-Actin was used as a loading control. Quantification of the CBS expression on mRNA level (RT-PCR method) and protein level (Western blotting method) was performed by densitometric analysis of the gels/blots and normalized to the internal loading control. The results are expressed as the mean ± SD of three or more independent experiments. Each mark represents p < 0.05 as followed: * versus REH; # versus DND-41; $ versus MOLT-4; & versus MV4-11; @ versus MOLM-14.

Figure 7

The expression on the mRNA (A) and protein (B) level of gamma-cystathionase in leukemia cell lines (REH, DND-41, MOLT-4, MV4-11, MOLM-14, K562). β-Actin was used as a loading control. Quantification of the CTH expression on mRNA level (RT-PCR method) and protein level (Western blotting method) was performed by densitometric analysis of the gels/blots and normalized to the internal loading control. The results are expressed as the mean ± SD of three or more independent experiments. Each mark represents p < 0.05 as followed: * versus REH; # versus DND-41; $ versus MOLT-4; & versus MV4-11; @ versus MOLM-14.