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Adenovirus-mediated siRNA targeting Bcl-xL inhibits proliferation, reduces invasion and enhances radiosensitivity of human colorectal cancer cells.

Yang J, Sun M, Zhang A, Lv C, De W, Wang Z - World J Surg Oncol (2011)

Bottom Line: Ad/shBcl-xL could significantly reduce the expression of Bcl-xL protein in CRC cells.Also, we showed that adenovirus-mediated siRNA targeting Bcl-xL could significantly inhibit proliferation and colony formation of CRC cells.Targeting Bcl-xL will be a promising strategy to inhibit the metastatic potential and reverse the radioresistance of human CRC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, Jiangsu, People's Republic of China.

ABSTRACT

Introduction: Bcl-xL, an important member of anti-apoptotic Bcl-2 family, plays critical roles in tumor progression and development. Previously, we have reported that overexpression of Bcl-xL was correlated with prognosis of colorectal cancer (CRC) patients. The aim of this study was to investigate the association of Bcl-xL expression with invasion and radiosensitivity of human CRC cells.

Methods: RT-PCR and Western blot assays were performed to determine the expression of Bcl-xL mRNA and protein in CRC cells and normal human intestinal epithelial cell line. Then, adenovirus-mediated RNA interference technique was employed to inhibit the expression of Bcl-xL gene in CRC cells. The proliferation of CRC cells was analyzed by MTT and colony formation assay. The migration and invasion of CRC cells was determined by wound-healing and tranwell invasion assays. Additionally, the in vitro and in vivo radiosensitivity of CRC cells was determined by clonogenic cell survival assay and murine xnograft model, respectively.

Results: The levels of Bcl-xL mRNA and protein expression were significantly higher in human CRC cells than in normal human intestinal epithelial cell line. Ad/shBcl-xL could significantly reduce the expression of Bcl-xL protein in CRC cells. Also, we showed that adenovirus-mediated siRNA targeting Bcl-xL could significantly inhibit proliferation and colony formation of CRC cells. Ad/shBcl-xL could significantly suppress migration and invasion of CRC cells. Moreover, Ad/shBcl-xL could enhance in vitro and in vivo radiosensitivity of CRC cells by increasing caspase-dependent apoptosis.

Conclusions: Targeting Bcl-xL will be a promising strategy to inhibit the metastatic potential and reverse the radioresistance of human CRC.

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Related in: MedlinePlus

Effects of Ad/shBcl-xL on the proferation and colony formation of CRC cells. A. The proliferation of LoVo cells was measured by MTT assay. The cell proliferation of LoVo infected with Ad/shBcl-xL was significantly inhibited in a time-dependent manner. B. The colony number was counted in three different wells at 14 days after seeded, and the averaged number was plotted. The experiments were repeated thrice and similar results were obtained. Representative data are shown. Each assay was performed at least in triplicate. *P < 0.05 vs mock.
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Figure 3: Effects of Ad/shBcl-xL on the proferation and colony formation of CRC cells. A. The proliferation of LoVo cells was measured by MTT assay. The cell proliferation of LoVo infected with Ad/shBcl-xL was significantly inhibited in a time-dependent manner. B. The colony number was counted in three different wells at 14 days after seeded, and the averaged number was plotted. The experiments were repeated thrice and similar results were obtained. Representative data are shown. Each assay was performed at least in triplicate. *P < 0.05 vs mock.

Mentions: Given that Ad/shBcl-xL could effectively inhibit Bcl-xL expression, its effects on the proliferation of CRC cells in vitro were determined. As shown in Figure 3A, compared with Ad/shcontrol or mock infection, Ad/shBcl-xL significantly decreased the viability of LoVo cells in a time-dependent manner, and the average proliferation inhibition rates at 3, 4, and 5 days were 39.2%, 42.9%, and 44.3%, respectively (P < 0.05). Additionally, Ad/shBcl-xL could significantly reduce the colony formation of LoVo cell by approximately by 35.4% (P < 0.01; Figure 3B). These results suggested that siRNA-mediated inhibition of Bcl-xL could suppress the proliferation of CRC cells.


Adenovirus-mediated siRNA targeting Bcl-xL inhibits proliferation, reduces invasion and enhances radiosensitivity of human colorectal cancer cells.

Yang J, Sun M, Zhang A, Lv C, De W, Wang Z - World J Surg Oncol (2011)

Effects of Ad/shBcl-xL on the proferation and colony formation of CRC cells. A. The proliferation of LoVo cells was measured by MTT assay. The cell proliferation of LoVo infected with Ad/shBcl-xL was significantly inhibited in a time-dependent manner. B. The colony number was counted in three different wells at 14 days after seeded, and the averaged number was plotted. The experiments were repeated thrice and similar results were obtained. Representative data are shown. Each assay was performed at least in triplicate. *P < 0.05 vs mock.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3198948&req=5

Figure 3: Effects of Ad/shBcl-xL on the proferation and colony formation of CRC cells. A. The proliferation of LoVo cells was measured by MTT assay. The cell proliferation of LoVo infected with Ad/shBcl-xL was significantly inhibited in a time-dependent manner. B. The colony number was counted in three different wells at 14 days after seeded, and the averaged number was plotted. The experiments were repeated thrice and similar results were obtained. Representative data are shown. Each assay was performed at least in triplicate. *P < 0.05 vs mock.
Mentions: Given that Ad/shBcl-xL could effectively inhibit Bcl-xL expression, its effects on the proliferation of CRC cells in vitro were determined. As shown in Figure 3A, compared with Ad/shcontrol or mock infection, Ad/shBcl-xL significantly decreased the viability of LoVo cells in a time-dependent manner, and the average proliferation inhibition rates at 3, 4, and 5 days were 39.2%, 42.9%, and 44.3%, respectively (P < 0.05). Additionally, Ad/shBcl-xL could significantly reduce the colony formation of LoVo cell by approximately by 35.4% (P < 0.01; Figure 3B). These results suggested that siRNA-mediated inhibition of Bcl-xL could suppress the proliferation of CRC cells.

Bottom Line: Ad/shBcl-xL could significantly reduce the expression of Bcl-xL protein in CRC cells.Also, we showed that adenovirus-mediated siRNA targeting Bcl-xL could significantly inhibit proliferation and colony formation of CRC cells.Targeting Bcl-xL will be a promising strategy to inhibit the metastatic potential and reverse the radioresistance of human CRC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, Jiangsu, People's Republic of China.

ABSTRACT

Introduction: Bcl-xL, an important member of anti-apoptotic Bcl-2 family, plays critical roles in tumor progression and development. Previously, we have reported that overexpression of Bcl-xL was correlated with prognosis of colorectal cancer (CRC) patients. The aim of this study was to investigate the association of Bcl-xL expression with invasion and radiosensitivity of human CRC cells.

Methods: RT-PCR and Western blot assays were performed to determine the expression of Bcl-xL mRNA and protein in CRC cells and normal human intestinal epithelial cell line. Then, adenovirus-mediated RNA interference technique was employed to inhibit the expression of Bcl-xL gene in CRC cells. The proliferation of CRC cells was analyzed by MTT and colony formation assay. The migration and invasion of CRC cells was determined by wound-healing and tranwell invasion assays. Additionally, the in vitro and in vivo radiosensitivity of CRC cells was determined by clonogenic cell survival assay and murine xnograft model, respectively.

Results: The levels of Bcl-xL mRNA and protein expression were significantly higher in human CRC cells than in normal human intestinal epithelial cell line. Ad/shBcl-xL could significantly reduce the expression of Bcl-xL protein in CRC cells. Also, we showed that adenovirus-mediated siRNA targeting Bcl-xL could significantly inhibit proliferation and colony formation of CRC cells. Ad/shBcl-xL could significantly suppress migration and invasion of CRC cells. Moreover, Ad/shBcl-xL could enhance in vitro and in vivo radiosensitivity of CRC cells by increasing caspase-dependent apoptosis.

Conclusions: Targeting Bcl-xL will be a promising strategy to inhibit the metastatic potential and reverse the radioresistance of human CRC.

Show MeSH
Related in: MedlinePlus