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Identification of the sAPRIL binding peptide and its growth inhibition effects in the colorectal cancer cells.

He XQ, Guan J, Liu F, Li J, He MR - PLoS ONE (2015)

Bottom Line: The effects of sAPRIL-BP on cell proliferation and cell cycle/apoptosis in vitro were evaluated using the CCK-8 assay and flow cytometry, respectively.The identified sAPRIL-BP suppressed tumor cell proliferation and cell cycle progression in LOVO cells in a dose-dependent manner.In vivo in a mouse colorectal challenge model, the sAPRIL-BP reduced the growth of tumor xenografts in nude mice by inhibiting proliferation and inducing apoptosis intratumorally.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Key Laboratory of Gastroenterology, Institute of Digestive Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China; Oncology Department, Wuzhou Red Cross Hospital, Wuzhou 543002, Guangxi Province, China.

ABSTRACT

Background: A proliferation-inducing ligand (APRIL) is a member of the tumor necrosis factor (TNF) super family. It binds to its specific receptors and is involved in multiple processes during tumorigenesis and tumor cells proliferation. High levels of APRIL expression are closely correlated to the growth, metastasis, and 5-FU drug resistance of colorectal cancer. The aim of this study was to identify a specific APRIL binding peptide (BP) able to block APRIL activity that could be used as a potential treatment for colorectal cancer.

Methods: A phage display library was used to identify peptides that bound selectively to soluble recombinant human APRIL (sAPRIL). The peptides with the highest binding affinity for sAPRIL were identified using ELISA. The effects of sAPRIL-BP on cell proliferation and cell cycle/apoptosis in vitro were evaluated using the CCK-8 assay and flow cytometry, respectively. An in vivo mouse model of colorectal cancer was used to determine the anti-tumor efficacy of the sAPRIL-BP.

Results: Three candidate peptides were characterized from eight phage clones with high binding affinity for sAPRIL. The peptide with the highest affinity was selected for further characterization. The identified sAPRIL-BP suppressed tumor cell proliferation and cell cycle progression in LOVO cells in a dose-dependent manner. In vivo in a mouse colorectal challenge model, the sAPRIL-BP reduced the growth of tumor xenografts in nude mice by inhibiting proliferation and inducing apoptosis intratumorally. Moreover, in an in vivo metastasis model, sAPRIL-BP reduced liver metastasis of colorectal cancer cells.

Conclusions: sAPRIL-BP significantly suppressed tumor growth in vitro and in vivo and might be a candidate for treating colorectal cancers that express high levels of APRIL.

No MeSH data available.


Related in: MedlinePlus

APRIL mRNA and protein expression in human colorectal cell lines.APRIL expression was assessed in five human colorectal cell lines (indicated) using RT-PCR (A) and Western Blotting (C). Representative gel images are shown. GAPDH was used as the internal control. The optical densities of the APRIL mRNA (B) and protein (D) bands were analyzed and normalized to the internal control. *P<0.05 compared to SW620, HT-29, or SW480.
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pone.0120564.g002: APRIL mRNA and protein expression in human colorectal cell lines.APRIL expression was assessed in five human colorectal cell lines (indicated) using RT-PCR (A) and Western Blotting (C). Representative gel images are shown. GAPDH was used as the internal control. The optical densities of the APRIL mRNA (B) and protein (D) bands were analyzed and normalized to the internal control. *P<0.05 compared to SW620, HT-29, or SW480.

Mentions: To evaluate the role of APRIL in colorectal cancer, APRIL expression was examined in five human colorectal cell lines by RT-PCR (Fig 2A and 2B) and western blotting (Fig 2C and 2D). The HCT116 and LOVO cell lines had significantly higher levels of APRIL mRNA (p<0.05) and protein (p<0.05) than the SW480, SW620, and HT29 cell lines. Therefore, the effect of sAPRIL-BP was evaluated in LOVO and HCT116 (APRILhigh) cells, and SW620 and HT-29 (APRILlow) cells. To determine whether there were dose effects, cell proliferation was measured at different concentrations of sAPRIL-BP. The rate of proliferation was significantly inhibited (p<0.05) by treatment with sAPRIL-BP in LOVO and HCT116 cells (Fig 3A) in a time- and dose-dependent manner. However, this was not the case in SW620 and HT-29 cells (Fig 3B). There results suggest that the anti-proliferative effects of sAPRIL-BP are specific for APRILhigh cells.


Identification of the sAPRIL binding peptide and its growth inhibition effects in the colorectal cancer cells.

He XQ, Guan J, Liu F, Li J, He MR - PLoS ONE (2015)

APRIL mRNA and protein expression in human colorectal cell lines.APRIL expression was assessed in five human colorectal cell lines (indicated) using RT-PCR (A) and Western Blotting (C). Representative gel images are shown. GAPDH was used as the internal control. The optical densities of the APRIL mRNA (B) and protein (D) bands were analyzed and normalized to the internal control. *P<0.05 compared to SW620, HT-29, or SW480.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4380366&req=5

pone.0120564.g002: APRIL mRNA and protein expression in human colorectal cell lines.APRIL expression was assessed in five human colorectal cell lines (indicated) using RT-PCR (A) and Western Blotting (C). Representative gel images are shown. GAPDH was used as the internal control. The optical densities of the APRIL mRNA (B) and protein (D) bands were analyzed and normalized to the internal control. *P<0.05 compared to SW620, HT-29, or SW480.
Mentions: To evaluate the role of APRIL in colorectal cancer, APRIL expression was examined in five human colorectal cell lines by RT-PCR (Fig 2A and 2B) and western blotting (Fig 2C and 2D). The HCT116 and LOVO cell lines had significantly higher levels of APRIL mRNA (p<0.05) and protein (p<0.05) than the SW480, SW620, and HT29 cell lines. Therefore, the effect of sAPRIL-BP was evaluated in LOVO and HCT116 (APRILhigh) cells, and SW620 and HT-29 (APRILlow) cells. To determine whether there were dose effects, cell proliferation was measured at different concentrations of sAPRIL-BP. The rate of proliferation was significantly inhibited (p<0.05) by treatment with sAPRIL-BP in LOVO and HCT116 cells (Fig 3A) in a time- and dose-dependent manner. However, this was not the case in SW620 and HT-29 cells (Fig 3B). There results suggest that the anti-proliferative effects of sAPRIL-BP are specific for APRILhigh cells.

Bottom Line: The effects of sAPRIL-BP on cell proliferation and cell cycle/apoptosis in vitro were evaluated using the CCK-8 assay and flow cytometry, respectively.The identified sAPRIL-BP suppressed tumor cell proliferation and cell cycle progression in LOVO cells in a dose-dependent manner.In vivo in a mouse colorectal challenge model, the sAPRIL-BP reduced the growth of tumor xenografts in nude mice by inhibiting proliferation and inducing apoptosis intratumorally.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Key Laboratory of Gastroenterology, Institute of Digestive Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China; Oncology Department, Wuzhou Red Cross Hospital, Wuzhou 543002, Guangxi Province, China.

ABSTRACT

Background: A proliferation-inducing ligand (APRIL) is a member of the tumor necrosis factor (TNF) super family. It binds to its specific receptors and is involved in multiple processes during tumorigenesis and tumor cells proliferation. High levels of APRIL expression are closely correlated to the growth, metastasis, and 5-FU drug resistance of colorectal cancer. The aim of this study was to identify a specific APRIL binding peptide (BP) able to block APRIL activity that could be used as a potential treatment for colorectal cancer.

Methods: A phage display library was used to identify peptides that bound selectively to soluble recombinant human APRIL (sAPRIL). The peptides with the highest binding affinity for sAPRIL were identified using ELISA. The effects of sAPRIL-BP on cell proliferation and cell cycle/apoptosis in vitro were evaluated using the CCK-8 assay and flow cytometry, respectively. An in vivo mouse model of colorectal cancer was used to determine the anti-tumor efficacy of the sAPRIL-BP.

Results: Three candidate peptides were characterized from eight phage clones with high binding affinity for sAPRIL. The peptide with the highest affinity was selected for further characterization. The identified sAPRIL-BP suppressed tumor cell proliferation and cell cycle progression in LOVO cells in a dose-dependent manner. In vivo in a mouse colorectal challenge model, the sAPRIL-BP reduced the growth of tumor xenografts in nude mice by inhibiting proliferation and inducing apoptosis intratumorally. Moreover, in an in vivo metastasis model, sAPRIL-BP reduced liver metastasis of colorectal cancer cells.

Conclusions: sAPRIL-BP significantly suppressed tumor growth in vitro and in vivo and might be a candidate for treating colorectal cancers that express high levels of APRIL.

No MeSH data available.


Related in: MedlinePlus