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Inhibition of c-myc expression in human rectal cancer cell line Colo320 by siRNA technique
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Abstract:
Objective: To inhibit c-myc gene expression in human rectal cancer cell line Colo320 by small interfering RNA technique, so as to assess the role of c-myc in Colo320 cells. Methods: The c-myc gene specific siRNA was designed and prepared by in vitro transcription. The prepared c-myc siRNA was transfected into Colo320 cells. After cultured for 48-96 hours, the cells were harvested. c-myc mRNA and protein level was monitored by fluorescence real time reverse transcription-poly merase chain reaction, c-myc protein expression was detected by Western blotting, and the cell proliferation activities were assayed by tetrazolium bromide (MTT) colorimetry and clone test. Results: Compared with control group, the mRNA of pGensil-c-myc-1, 2, 3, and 4 in Colo320 cells was obviously decreased in c-myc siRNA transfected cells. C-myc protein expression was also decreased obviously. MTT assay and clone test showed that c-myc siRNA apparently slowed down the proliferation of Colo320 cells. Conclusion: Our results suggest that RNA interference exists in Colo320 cell line; c-myc siRNA can specifically inhibit the expression of c-myc gene in Colo320 cells, subsequently inhibits the proliferation of Colo320 cells.
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