[Expression of ICAT and Wnt signaling-related proteins in the monocytic differentiation of HL-60 cells induced by a new steroidal drug NSC67657].

Abstract

OBJECTIVE:

To investigate the expression of mRNA and proteins of β-catenin, TCF-4 (ICAT) and Wnt signaling pathway-related genes in the monocytic differentiation of acute myeloid leukemia HL-60 cells induced by a new steroidal drug NSC67657.

METHODS:

Wright's staining and α-NBE staining were used to observe the differentiation of HL-60 cells after 5 days of 10 μmol/L NSC67657 treatment. Flow cytometry (FCM) was used to detect the differentiation and cell cycles. The expressions of mRNA and proteins of ICAT and Wnt signaling pathway-related factors, including β-catenin, TCF-4, c-myc, cyclin D1 and TCF-1 before and after differentiation, were detected by RT-PCR and Western blot.

RESULTS:

Morphological observation showed that NSC67657 induced monocytic differentiation of HL-60 cells. At 5 days after 10 μmol/L NSC67657 treatment, the number of CD14(+) HL-60 cells was (94.37±2.84)%, significantly higher than the (1.31±0.09)% in control group (P<0.01). The flow cytometry assay revealed that NSC67657 induced (76.46±2.83)% of G1/G0 phase arrest, significantly higher than that of (59.40±5.42)% in the control group (P<0.05), while the S phase cells were of (18.76±0.98)%, significantly lower than that of (34.38±2.61) % in the control group (P<0.05). The NSC67657 treatment also up-regulated the expression of ICAT mRNA and protein, and down-regulated the expression of β-catenin mRNA and protin (P<0.01 for all). However, the nuclear expression of β-catenin was down-regulated (P<0.01). The NSC67657 treatment induced nonsignificant alterations of TCF-4 mRNA, total protein and nuclear protein in the HL-60 cells (P>0.05 for all). The target genes of Wnt signaling pathway, including c-myc, cyclinD1 and TCF-1 mRNA and proteins in the HL-60 cells were significantly down-regulated after NSC67657 treatment (P<0.05).

CONCLUSIONS:

The new steroidal drug NSC67657 induces monocytic differentiation of HL-60 cells, and down-regulates the expression of β-catenin and target genes of Wnt signaling pathway. These results indicate that Wnt signaling pathway may be directly or indirectly involved in the monocytic differentiation process of HL-60 cells.

• Authors: J S Wang, W J Wang, T Wang, Y Zhang
• Journal: Zhonghua zhong liu za zhi [Chinese journal of oncology] (Zhonghua Zhong Liu Za Zhi) Vol. 38 Issue 4 Pg. 246-51 (Apr 2016) ISSN: 0253-3766 [Print] China
• PMID: 27087369 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• (4-(10, 13-dimethyl-3-methylsulfonyloxy-2,3,4,5,6,7,8,9,11,12,14,15,16, 17-tetradecahydro-1H-cyclopenta(a)phenanthren-17-yl)pentyl) methanesulfonate
• Adaptor Proteins, Signal Transducing
• CTNNBIP1 protein, human
• Intracellular Signaling Peptides and Proteins
• Mesylates
• RNA, Messenger
• Steroids
• T Cell Transcription Factor 1
• TCF7 protein, human
• Wnt Proteins
• beta Catenin
• Cyclin D1

Topics

• Adaptor Proteins, Signal Transducing
• Cell Cycle
• Cell Differentiation
• Cyclin D1 (genetics, metabolism)
• Down-Regulation
• HL-60 Cells (cytology, drug effects, metabolism)
• Humans
• Intracellular Signaling Peptides and Proteins (genetics, metabolism)
• Mesylates (pharmacology)
• RNA, Messenger (metabolism)
• Signal Transduction (drug effects)
• Steroids (pharmacology)
• T Cell Transcription Factor 1 (genetics, metabolism)
• Transfection
• Wnt Proteins (metabolism)
• beta Catenin (genetics, metabolism)