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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

The expression of Bcl-xL mRNA and protein in four human CRC cell lines (SW480, HT-29, LoVo and Colo320) and one intestinal epithelial cell line (HIEC) by RT-PCR (A) and Western blot assay (B). GAPDH was used as an internal control.
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Figure 1: The expression of Bcl-xL mRNA and protein in four human CRC cell lines (SW480, HT-29, LoVo and Colo320) and one intestinal epithelial cell line (HIEC) by RT-PCR (A) and Western blot assay (B). GAPDH was used as an internal control.

Mentions: Semi-quantitative RT-PCR and Western blot assays were done to determine the expression of Bcl-xL mRNA and protein in four human CRC cell lines (SW480, HT-29, Colo320 and LoVo) and a human intestinal epithelial cell line (HIEC). The expression levels of Bcl-xL mRNA and protein were obviously higher in human CRC cell lines than in human intestinal epithelial cell line (Figure 1A and 1B). Among CRC cell lines, the expression levels of Bcl-xL mRNA and protein shows difference. Meanwhile, in our previous study, we also showed that the average level of Bcl-xL gene in colorectal cancer tissues was significantly higher than that in corresponding nontumor colon tissues at both transcriptional and translational levels. Thus, it was concluded that the overexpression of Bcl-xL might play major roles in CRC tumorigenesis and progression.


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)

The expression of Bcl-xL mRNA and protein in four human CRC cell lines (SW480, HT-29, LoVo and Colo320) and one intestinal epithelial cell line (HIEC) by RT-PCR (A) and Western blot assay (B). GAPDH was used as an internal control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: The expression of Bcl-xL mRNA and protein in four human CRC cell lines (SW480, HT-29, LoVo and Colo320) and one intestinal epithelial cell line (HIEC) by RT-PCR (A) and Western blot assay (B). GAPDH was used as an internal control.
Mentions: Semi-quantitative RT-PCR and Western blot assays were done to determine the expression of Bcl-xL mRNA and protein in four human CRC cell lines (SW480, HT-29, Colo320 and LoVo) and a human intestinal epithelial cell line (HIEC). The expression levels of Bcl-xL mRNA and protein were obviously higher in human CRC cell lines than in human intestinal epithelial cell line (Figure 1A and 1B). Among CRC cell lines, the expression levels of Bcl-xL mRNA and protein shows difference. Meanwhile, in our previous study, we also showed that the average level of Bcl-xL gene in colorectal cancer tissues was significantly higher than that in corresponding nontumor colon tissues at both transcriptional and translational levels. Thus, it was concluded that the overexpression of Bcl-xL might play major roles in CRC tumorigenesis and progression.

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