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Involvement of calreticulin in cell proliferation, invasion and differentiation in diallyl disulfide-treated HL-60 cells

View Article: PubMed Central - PubMed

ABSTRACT

Diallyl disulfide (DADS) has shown potential as a therapeutic agent in various cancers. Previously, calreticulin (CRT) was found to be downregulated in differentiated HL-60 cells treated with DADS. The present study investigated the role of CRT proteins in DADS-induced proliferation, invasion and differentiation in HL-60 cells. The present study demonstrated that DADS treatment significantly changed the morphology of HL-60 cells and caused the significant time-dependent downregulation of CRT. Small interfering RNA (siRNA)-mediated knockdown of CRT expression significantly inhibited proliferation, decreased invasion ability, increased the expression of cluster of differentiation (CD)11b and reduced the expression of CD33 in DADS-treated HL-60 cells. DADS also significantly affected cell proliferation, invasion and differentiation in CRT-overexpressed HL-60 cells. Nitroblue tetrazolium (NBT) reduction assays showed decreased NBT reduction activity in the CRT overexpression group and increased NBT reduction in the CRT siRNA group. Following treatment with DADS, the NBT reduction abilities in all groups were increased. In conclusion, the present study clearly demonstrates the downregulation of CRT during DADS-induced differentiation in HL-60 cells and indicates that CRT is involved in cell proliferation, invasion and differentiation in DADS-treated HL-60 cells.

No MeSH data available.


Related in: MedlinePlus

Role of CRT in cell invasion of HL-60 cells induced by DADS. (A) Representative experiment of Transwell invasion assays (magnification, ×200): 1, HL-60 cells; 2, CRT-overexpressing HL-60 cells; 3, CRT-downregulated HL-60 cells; 4, DADS-treated HL-60 cells; 5, DADS-treated CRT-overexpressing HL-60 cells; 6, DADS-treated CRT-downregulated HL-60 cells. (B) Values are mean ± standard from three independent experiments. *P<0.05 vs. control cells. CRT, calreticulin; DADS, diallyl disulfide.
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f3-ol-0-0-4850: Role of CRT in cell invasion of HL-60 cells induced by DADS. (A) Representative experiment of Transwell invasion assays (magnification, ×200): 1, HL-60 cells; 2, CRT-overexpressing HL-60 cells; 3, CRT-downregulated HL-60 cells; 4, DADS-treated HL-60 cells; 5, DADS-treated CRT-overexpressing HL-60 cells; 6, DADS-treated CRT-downregulated HL-60 cells. (B) Values are mean ± standard from three independent experiments. *P<0.05 vs. control cells. CRT, calreticulin; DADS, diallyl disulfide.

Mentions: The present study next investigated the role of CRT in cell invasion induced by DADS treatment. Transwell Matrigel invasion assays showed that the number of HL-60 cells that penetrated through the Matrigel in the non-transfected group was 62±6.37, compared with 83±11.39 in the CRT overexpression group and 39±2.43 in the CRT siRNA group (Fig. 3A and B). These results indicate that CRT overexpression can increase the ability of invasion in HL-60 cells and CRT siRNA transfection can decrease the ability of invasion. Following treatment with DADS for 48 h, the number of HL-60 cells that penetrated through the Matrigel was 36.2±7.59 in the non-transfected group, compared with 45.6±4.10 and 23.45±3.86 in the CRT overexpressing and downregulated groups, respectively. Cellular invasion in the DADS treated groups was decreased compared with the untreated groups (P=0.034). These results indicate that CRT is involved in cell invasion in DADS-induced HL-60 cells.


Involvement of calreticulin in cell proliferation, invasion and differentiation in diallyl disulfide-treated HL-60 cells
Role of CRT in cell invasion of HL-60 cells induced by DADS. (A) Representative experiment of Transwell invasion assays (magnification, ×200): 1, HL-60 cells; 2, CRT-overexpressing HL-60 cells; 3, CRT-downregulated HL-60 cells; 4, DADS-treated HL-60 cells; 5, DADS-treated CRT-overexpressing HL-60 cells; 6, DADS-treated CRT-downregulated HL-60 cells. (B) Values are mean ± standard from three independent experiments. *P<0.05 vs. control cells. CRT, calreticulin; DADS, diallyl disulfide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4998039&req=5

f3-ol-0-0-4850: Role of CRT in cell invasion of HL-60 cells induced by DADS. (A) Representative experiment of Transwell invasion assays (magnification, ×200): 1, HL-60 cells; 2, CRT-overexpressing HL-60 cells; 3, CRT-downregulated HL-60 cells; 4, DADS-treated HL-60 cells; 5, DADS-treated CRT-overexpressing HL-60 cells; 6, DADS-treated CRT-downregulated HL-60 cells. (B) Values are mean ± standard from three independent experiments. *P<0.05 vs. control cells. CRT, calreticulin; DADS, diallyl disulfide.
Mentions: The present study next investigated the role of CRT in cell invasion induced by DADS treatment. Transwell Matrigel invasion assays showed that the number of HL-60 cells that penetrated through the Matrigel in the non-transfected group was 62±6.37, compared with 83±11.39 in the CRT overexpression group and 39±2.43 in the CRT siRNA group (Fig. 3A and B). These results indicate that CRT overexpression can increase the ability of invasion in HL-60 cells and CRT siRNA transfection can decrease the ability of invasion. Following treatment with DADS for 48 h, the number of HL-60 cells that penetrated through the Matrigel was 36.2±7.59 in the non-transfected group, compared with 45.6±4.10 and 23.45±3.86 in the CRT overexpressing and downregulated groups, respectively. Cellular invasion in the DADS treated groups was decreased compared with the untreated groups (P=0.034). These results indicate that CRT is involved in cell invasion in DADS-induced HL-60 cells.

View Article: PubMed Central - PubMed

ABSTRACT

Diallyl disulfide (DADS) has shown potential as a therapeutic agent in various cancers. Previously, calreticulin (CRT) was found to be downregulated in differentiated HL-60 cells treated with DADS. The present study investigated the role of CRT proteins in DADS-induced proliferation, invasion and differentiation in HL-60 cells. The present study demonstrated that DADS treatment significantly changed the morphology of HL-60 cells and caused the significant time-dependent downregulation of CRT. Small interfering RNA (siRNA)-mediated knockdown of CRT expression significantly inhibited proliferation, decreased invasion ability, increased the expression of cluster of differentiation (CD)11b and reduced the expression of CD33 in DADS-treated HL-60 cells. DADS also significantly affected cell proliferation, invasion and differentiation in CRT-overexpressed HL-60 cells. Nitroblue tetrazolium (NBT) reduction assays showed decreased NBT reduction activity in the CRT overexpression group and increased NBT reduction in the CRT siRNA group. Following treatment with DADS, the NBT reduction abilities in all groups were increased. In conclusion, the present study clearly demonstrates the downregulation of CRT during DADS-induced differentiation in HL-60 cells and indicates that CRT is involved in cell proliferation, invasion and differentiation in DADS-treated HL-60 cells.

No MeSH data available.


Related in: MedlinePlus