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Bifunctional fused polypeptide inhibits the growth and metastasis of breast cancer.

Liang AL, Qian HL, Zhang TT, Zhou N, Wang HJ, Men XT, Qi W, Zhang PP, Fu M, Liang X, Lin C, Liu YJ - Drug Des Devel Ther (2015)

Bottom Line: This results in a potent inhibitory effect of fused TAT-DV1-BH3 polypeptide on tumor growth and metastasis in nude mice bearing established MDA-MB-231 tumors.Notably, the DV1-mediated inhibition of the stromal-derived factor-1/CXCR4 pathway contributed to the antimetastasis effect, evident from the reduction in the level of phosphoinositide 3 kinase and matrix metalloproteinase 9 in MDA-MB-231 cells.Collectively, these results indicate that the apoptosis-inducing effect and migration- and invasion-suppressing effect explain the tumor regression and metastasis inhibition in vivo, with the involvement of caspase- and CXCR4-mediated signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Medical Molecular Diagnostics Key Laboratory of Guangdong, Guangdong Medical University, Dongguan, Guangdong, People's Republic of China ; Department of Biochemistry and Molecular Biology, Guangdong Medical University, Dongguan, Guangdong, People's Republic of China ; Department of Clinical Biochemistry, Guangdong Medical University, Dongguan, Guangdong, People's Republic of China.

ABSTRACT
Breast cancer is the most common cancer and the leading cause of cancer-related death among women worldwide, with urgent need to develop new therapeutics. Targeted therapy is a promising strategy for breast cancer therapy. Stromal-derived factor-1/CXC chemokine receptor 4 (CXCR4) has been implicated in the metastasis of breast cancer, which renders it to be therapeutic target. This study aimed to evaluate the anticancer effect of fused TAT- DV1-BH3 polypeptide, an antagonist of CXCR4, and investigate the underlying mechanism for the cancer cell-killing effect in the treatment of breast cancer in vitro and in vivo. This results in a potent inhibitory effect of fused TAT-DV1-BH3 polypeptide on tumor growth and metastasis in nude mice bearing established MDA-MB-231 tumors. Fused TAT-DV1-BH3 polypeptide inhibited the proliferation of MDA-MB-231 and MCF-7 cells but did not affect that of HEK-293 cells. The fused TAT-DV1-BH3 polypeptide colocalized with mitochondria and exhibited a proapoptotic effect through the regulation of caspase-9 and -3. Furthermore, the fused TAT-DV1-BH3 polypeptide suppressed the migration and invasion of the highly metastatic breast cancer cell line MDA-MB-231 in a concentration-dependent manner. Notably, the DV1-mediated inhibition of the stromal-derived factor-1/CXCR4 pathway contributed to the antimetastasis effect, evident from the reduction in the level of phosphoinositide 3 kinase and matrix metalloproteinase 9 in MDA-MB-231 cells. Collectively, these results indicate that the apoptosis-inducing effect and migration- and invasion-suppressing effect explain the tumor regression and metastasis inhibition in vivo, with the involvement of caspase- and CXCR4-mediated signaling pathway. The data suggest that the fused TAT-DV1-BH3 polypeptide is a promising agent for the treatment of breast cancer, and more studies are warranted to fully elucidate the therapeutic targets and molecular mechanism.

No MeSH data available.


Related in: MedlinePlus

The colocalization of DV1–BH3, TAT–DV1, and TAT–DV1–BH3 in MDA-MB-231, MCF-7, and HEK-293 cells.Notes: The fused polypeptides and the mitochondria were observed using laser scanning confocal microscopy. MDA-MB-231 (A), MCF-7 (B), and HEK-293 (C) cells were treated with 40 μM DV1–BH3, TAT–DV1, or TAT–DV1–BH3 for 1 hour and washed with PBS, then the cells were stained with Mito Tracker Red CMXRos. The green fluorescence was from the polypeptides and the red fluorescence was from the mitochondria. Scale bar, 10 μm.Abbreviations: BH3, Bcl-2 homology 3; TAT, transactivator of transcription; PBS, phosphate-buffered saline.
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f2-dddt-9-5671: The colocalization of DV1–BH3, TAT–DV1, and TAT–DV1–BH3 in MDA-MB-231, MCF-7, and HEK-293 cells.Notes: The fused polypeptides and the mitochondria were observed using laser scanning confocal microscopy. MDA-MB-231 (A), MCF-7 (B), and HEK-293 (C) cells were treated with 40 μM DV1–BH3, TAT–DV1, or TAT–DV1–BH3 for 1 hour and washed with PBS, then the cells were stained with Mito Tracker Red CMXRos. The green fluorescence was from the polypeptides and the red fluorescence was from the mitochondria. Scale bar, 10 μm.Abbreviations: BH3, Bcl-2 homology 3; TAT, transactivator of transcription; PBS, phosphate-buffered saline.

Mentions: To observe the cellular localization of these three polypeptides, laser scanning confocal microscopic examination was performed to measure the green fluorescence from the FITC-tagged polypeptides and the red fluorescence from mitochondria stained with the specific tracking dye MitoTracker® Red CMXRos. MDA-MB-231, MCF-7, and HEK-293 cells were incubated with 40 μM TAT–DV1–BH3, TAT–DV1, or DV1–BH3 for 1 hour, and the fluorescence was examined (Figure 2A–C). The fused TAT–DV1 and TAT–DV1–BH3 polypeptides entered into cells and largely distributed in the cytoplasm, due to the TAT sequence (Figure 2A–C). Notably, these two fused polypeptides containing TAT were found to colocalize with mitochondria in MDA-MB-231, MCF-7, and HEK-293 cells (Figure 2A–C). However, due to the lack of TAT sequence, the DV1–BH3 polypeptide was unable to enter into cells after co-culturing with cells for 1 hour, evident from the single observation of red fluorescence (Figure 2A–C).


Bifunctional fused polypeptide inhibits the growth and metastasis of breast cancer.

Liang AL, Qian HL, Zhang TT, Zhou N, Wang HJ, Men XT, Qi W, Zhang PP, Fu M, Liang X, Lin C, Liu YJ - Drug Des Devel Ther (2015)

The colocalization of DV1–BH3, TAT–DV1, and TAT–DV1–BH3 in MDA-MB-231, MCF-7, and HEK-293 cells.Notes: The fused polypeptides and the mitochondria were observed using laser scanning confocal microscopy. MDA-MB-231 (A), MCF-7 (B), and HEK-293 (C) cells were treated with 40 μM DV1–BH3, TAT–DV1, or TAT–DV1–BH3 for 1 hour and washed with PBS, then the cells were stained with Mito Tracker Red CMXRos. The green fluorescence was from the polypeptides and the red fluorescence was from the mitochondria. Scale bar, 10 μm.Abbreviations: BH3, Bcl-2 homology 3; TAT, transactivator of transcription; PBS, phosphate-buffered saline.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4621185&req=5

f2-dddt-9-5671: The colocalization of DV1–BH3, TAT–DV1, and TAT–DV1–BH3 in MDA-MB-231, MCF-7, and HEK-293 cells.Notes: The fused polypeptides and the mitochondria were observed using laser scanning confocal microscopy. MDA-MB-231 (A), MCF-7 (B), and HEK-293 (C) cells were treated with 40 μM DV1–BH3, TAT–DV1, or TAT–DV1–BH3 for 1 hour and washed with PBS, then the cells were stained with Mito Tracker Red CMXRos. The green fluorescence was from the polypeptides and the red fluorescence was from the mitochondria. Scale bar, 10 μm.Abbreviations: BH3, Bcl-2 homology 3; TAT, transactivator of transcription; PBS, phosphate-buffered saline.
Mentions: To observe the cellular localization of these three polypeptides, laser scanning confocal microscopic examination was performed to measure the green fluorescence from the FITC-tagged polypeptides and the red fluorescence from mitochondria stained with the specific tracking dye MitoTracker® Red CMXRos. MDA-MB-231, MCF-7, and HEK-293 cells were incubated with 40 μM TAT–DV1–BH3, TAT–DV1, or DV1–BH3 for 1 hour, and the fluorescence was examined (Figure 2A–C). The fused TAT–DV1 and TAT–DV1–BH3 polypeptides entered into cells and largely distributed in the cytoplasm, due to the TAT sequence (Figure 2A–C). Notably, these two fused polypeptides containing TAT were found to colocalize with mitochondria in MDA-MB-231, MCF-7, and HEK-293 cells (Figure 2A–C). However, due to the lack of TAT sequence, the DV1–BH3 polypeptide was unable to enter into cells after co-culturing with cells for 1 hour, evident from the single observation of red fluorescence (Figure 2A–C).

Bottom Line: This results in a potent inhibitory effect of fused TAT-DV1-BH3 polypeptide on tumor growth and metastasis in nude mice bearing established MDA-MB-231 tumors.Notably, the DV1-mediated inhibition of the stromal-derived factor-1/CXCR4 pathway contributed to the antimetastasis effect, evident from the reduction in the level of phosphoinositide 3 kinase and matrix metalloproteinase 9 in MDA-MB-231 cells.Collectively, these results indicate that the apoptosis-inducing effect and migration- and invasion-suppressing effect explain the tumor regression and metastasis inhibition in vivo, with the involvement of caspase- and CXCR4-mediated signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Medical Molecular Diagnostics Key Laboratory of Guangdong, Guangdong Medical University, Dongguan, Guangdong, People's Republic of China ; Department of Biochemistry and Molecular Biology, Guangdong Medical University, Dongguan, Guangdong, People's Republic of China ; Department of Clinical Biochemistry, Guangdong Medical University, Dongguan, Guangdong, People's Republic of China.

ABSTRACT
Breast cancer is the most common cancer and the leading cause of cancer-related death among women worldwide, with urgent need to develop new therapeutics. Targeted therapy is a promising strategy for breast cancer therapy. Stromal-derived factor-1/CXC chemokine receptor 4 (CXCR4) has been implicated in the metastasis of breast cancer, which renders it to be therapeutic target. This study aimed to evaluate the anticancer effect of fused TAT- DV1-BH3 polypeptide, an antagonist of CXCR4, and investigate the underlying mechanism for the cancer cell-killing effect in the treatment of breast cancer in vitro and in vivo. This results in a potent inhibitory effect of fused TAT-DV1-BH3 polypeptide on tumor growth and metastasis in nude mice bearing established MDA-MB-231 tumors. Fused TAT-DV1-BH3 polypeptide inhibited the proliferation of MDA-MB-231 and MCF-7 cells but did not affect that of HEK-293 cells. The fused TAT-DV1-BH3 polypeptide colocalized with mitochondria and exhibited a proapoptotic effect through the regulation of caspase-9 and -3. Furthermore, the fused TAT-DV1-BH3 polypeptide suppressed the migration and invasion of the highly metastatic breast cancer cell line MDA-MB-231 in a concentration-dependent manner. Notably, the DV1-mediated inhibition of the stromal-derived factor-1/CXCR4 pathway contributed to the antimetastasis effect, evident from the reduction in the level of phosphoinositide 3 kinase and matrix metalloproteinase 9 in MDA-MB-231 cells. Collectively, these results indicate that the apoptosis-inducing effect and migration- and invasion-suppressing effect explain the tumor regression and metastasis inhibition in vivo, with the involvement of caspase- and CXCR4-mediated signaling pathway. The data suggest that the fused TAT-DV1-BH3 polypeptide is a promising agent for the treatment of breast cancer, and more studies are warranted to fully elucidate the therapeutic targets and molecular mechanism.

No MeSH data available.


Related in: MedlinePlus