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microRNA let-7c is essential for the anisomycin-elicited apoptosis in Jurkat T cells by linking JNK1/2 to AP-1/STAT1/STAT3 signaling.

Zhou Z, Lu X, Wang J, Xiao J, Liu J, Xing F - Sci Rep (2016)

Bottom Line: The results showed that anisomycin sufficiently promoted the apoptosis in human leukemic Jurkat T cells at a quite low dose. microRNA let-7c (let-7c) contributed to the anisomycin-induced apoptosis, which could be abrogated by the inactivation of JNK signaling.The let-7c deficiency reduced the AP-1, STAT1 and Bim activities, and enhanced the STAT3 and Bcl-xL, alleviating the anisomycin-induced apoptosis.This provides a novel insight into the mechanism by which anisomycin leads to the tumor cell apoptosis, potentially laying the foundations for its development and clinical application.

View Article: PubMed Central - PubMed

Affiliation: Institute of Tissue Transplantation and Immunology, Department of Immunobiology, Jinan University, Guangzhou 510632, China.

ABSTRACT
Anisomycin, an antibiotic produced by Streptomyces griseolus, strongly induces apoptosis in various tumor cells in vitro, superior dramatically to adriamycin. The present study aims to elucidate its detailed mechanistic process. The results showed that anisomycin sufficiently promoted the apoptosis in human leukemic Jurkat T cells at a quite low dose. microRNA let-7c (let-7c) contributed to the anisomycin-induced apoptosis, which could be abrogated by the inactivation of JNK signaling. The let-7c over-expression and the addition of its mimics facilitated the activation of AP-1, STAT1 and Bim by linking JNK1/2 to AP-1/STAT1, but rather inhibited the activation of STAT3 and Bcl-xL by connecting JNK1/2 to STAT3, followed by the augmented apoptosis in the cells. The let-7c deficiency reduced the AP-1, STAT1 and Bim activities, and enhanced the STAT3 and Bcl-xL, alleviating the anisomycin-induced apoptosis. The knockdown of the bim gene repressed the anisomycin-boosted apoptosis through the attenuation of the active Bak and Bax. The findings indicate for the first time that miR let-7c is essential for the anisomycin-triggered apoptosis by linking JNK1/2 to AP-1/STAT1/STAT3/Bim/Bcl-xL/Bax/Bak signaling. This provides a novel insight into the mechanism by which anisomycin leads to the tumor cell apoptosis, potentially laying the foundations for its development and clinical application.

No MeSH data available.


Related in: MedlinePlus

miRNA let-7c overexpression facilitates the anisomycin-stimulated apoptosis in Jurkat T cells.Jurkat T cells were electro-transfected with 2 μg of the let-7c overexpression vector with GFP marker (let-7c) or 2 μg of the miR-Negative Control vector (mock) for 24 h or 48 h. (A) GFP expression in Jurkat T cells. (B) The expression of miR let-7c in the transfected cells was measured by real-time qPCR. (C) The levels of phospho-c-jun, phospho-STAT1 and phospho-STAT3 were analyzed by Western blot. (D) The apoptotic rate of the treated cells was assayed by flow cytometry. The data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 vs. the untreated control, #p < 0.05, ##p < 0.01 vs. the let-7c overexpression group.
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f5: miRNA let-7c overexpression facilitates the anisomycin-stimulated apoptosis in Jurkat T cells.Jurkat T cells were electro-transfected with 2 μg of the let-7c overexpression vector with GFP marker (let-7c) or 2 μg of the miR-Negative Control vector (mock) for 24 h or 48 h. (A) GFP expression in Jurkat T cells. (B) The expression of miR let-7c in the transfected cells was measured by real-time qPCR. (C) The levels of phospho-c-jun, phospho-STAT1 and phospho-STAT3 were analyzed by Western blot. (D) The apoptotic rate of the treated cells was assayed by flow cytometry. The data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 vs. the untreated control, #p < 0.05, ##p < 0.01 vs. the let-7c overexpression group.

Mentions: To further explore the role of the miR let-7c augment in the anisomycin-treated cells, an overexpressed vector containing miR let-7c was electro-transfected into Jurkat T cells with the transfection rate of 60–70% (Fig. 5A). Consequently, miR let-7c facilitated the significant enhancement of AP-1 and STAT1 in the anisomycin-induced apoptosis. On the contrary, STAT3 was suppressed with the enhancement of miR let-7c (Fig. 5B,C). The foregoing alterations led to the occurrence of the cell apoptosis (Fig. 5D). These results suggest that the miRNA let-7c regulates AP-1/STAT1/STAT3 to mediate the anisomycin-induced apoptosis.


microRNA let-7c is essential for the anisomycin-elicited apoptosis in Jurkat T cells by linking JNK1/2 to AP-1/STAT1/STAT3 signaling.

Zhou Z, Lu X, Wang J, Xiao J, Liu J, Xing F - Sci Rep (2016)

miRNA let-7c overexpression facilitates the anisomycin-stimulated apoptosis in Jurkat T cells.Jurkat T cells were electro-transfected with 2 μg of the let-7c overexpression vector with GFP marker (let-7c) or 2 μg of the miR-Negative Control vector (mock) for 24 h or 48 h. (A) GFP expression in Jurkat T cells. (B) The expression of miR let-7c in the transfected cells was measured by real-time qPCR. (C) The levels of phospho-c-jun, phospho-STAT1 and phospho-STAT3 were analyzed by Western blot. (D) The apoptotic rate of the treated cells was assayed by flow cytometry. The data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 vs. the untreated control, #p < 0.05, ##p < 0.01 vs. the let-7c overexpression group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834478&req=5

f5: miRNA let-7c overexpression facilitates the anisomycin-stimulated apoptosis in Jurkat T cells.Jurkat T cells were electro-transfected with 2 μg of the let-7c overexpression vector with GFP marker (let-7c) or 2 μg of the miR-Negative Control vector (mock) for 24 h or 48 h. (A) GFP expression in Jurkat T cells. (B) The expression of miR let-7c in the transfected cells was measured by real-time qPCR. (C) The levels of phospho-c-jun, phospho-STAT1 and phospho-STAT3 were analyzed by Western blot. (D) The apoptotic rate of the treated cells was assayed by flow cytometry. The data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 vs. the untreated control, #p < 0.05, ##p < 0.01 vs. the let-7c overexpression group.
Mentions: To further explore the role of the miR let-7c augment in the anisomycin-treated cells, an overexpressed vector containing miR let-7c was electro-transfected into Jurkat T cells with the transfection rate of 60–70% (Fig. 5A). Consequently, miR let-7c facilitated the significant enhancement of AP-1 and STAT1 in the anisomycin-induced apoptosis. On the contrary, STAT3 was suppressed with the enhancement of miR let-7c (Fig. 5B,C). The foregoing alterations led to the occurrence of the cell apoptosis (Fig. 5D). These results suggest that the miRNA let-7c regulates AP-1/STAT1/STAT3 to mediate the anisomycin-induced apoptosis.

Bottom Line: The results showed that anisomycin sufficiently promoted the apoptosis in human leukemic Jurkat T cells at a quite low dose. microRNA let-7c (let-7c) contributed to the anisomycin-induced apoptosis, which could be abrogated by the inactivation of JNK signaling.The let-7c deficiency reduced the AP-1, STAT1 and Bim activities, and enhanced the STAT3 and Bcl-xL, alleviating the anisomycin-induced apoptosis.This provides a novel insight into the mechanism by which anisomycin leads to the tumor cell apoptosis, potentially laying the foundations for its development and clinical application.

View Article: PubMed Central - PubMed

Affiliation: Institute of Tissue Transplantation and Immunology, Department of Immunobiology, Jinan University, Guangzhou 510632, China.

ABSTRACT
Anisomycin, an antibiotic produced by Streptomyces griseolus, strongly induces apoptosis in various tumor cells in vitro, superior dramatically to adriamycin. The present study aims to elucidate its detailed mechanistic process. The results showed that anisomycin sufficiently promoted the apoptosis in human leukemic Jurkat T cells at a quite low dose. microRNA let-7c (let-7c) contributed to the anisomycin-induced apoptosis, which could be abrogated by the inactivation of JNK signaling. The let-7c over-expression and the addition of its mimics facilitated the activation of AP-1, STAT1 and Bim by linking JNK1/2 to AP-1/STAT1, but rather inhibited the activation of STAT3 and Bcl-xL by connecting JNK1/2 to STAT3, followed by the augmented apoptosis in the cells. The let-7c deficiency reduced the AP-1, STAT1 and Bim activities, and enhanced the STAT3 and Bcl-xL, alleviating the anisomycin-induced apoptosis. The knockdown of the bim gene repressed the anisomycin-boosted apoptosis through the attenuation of the active Bak and Bax. The findings indicate for the first time that miR let-7c is essential for the anisomycin-triggered apoptosis by linking JNK1/2 to AP-1/STAT1/STAT3/Bim/Bcl-xL/Bax/Bak signaling. This provides a novel insight into the mechanism by which anisomycin leads to the tumor cell apoptosis, potentially laying the foundations for its development and clinical application.

No MeSH data available.


Related in: MedlinePlus