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Archive > Volume 24 Issue 2 > 2017,24(2):168-173. DOI:10.3872/j.issn.1007-385X.2017.02.011 Prev Next
Clinical Research
Bcl-3 expression on the malignant biologic behavior of colorectal cancer HCT 116 cells
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Abstract:
Objective:To explore Bcl-3 protein expression in human colorectal cancer and the effects of Bcl-3 gene silencing on proliferation and apoptosis of colorectal cancer HCT 116 cell line. Methods: 86 colorectal cancer tissues and the corresponding adjacent tissues from patients treated in Kunshan Hospital Affiliated to Jiangsu University between January 2012 and December 2015 were collected to analyze the correlation between Bcl-3 expression and clinicopathological features of colorectal cancer. Immunohistochemistry was used to detect Bcl-3 protein expression in 86 samples. HCT 116 cells were cultivated and divided into three groups: blank control group, control siRNA group (transfecting with control siRNA) and Bcl-3 siRNA (transfecting with Bcl-3 siRNA); and then Western blotting was used to determine Bcl-3 expression to verify the transfection efficiency. In addition, MTT, plate clone formation assay, Flow cytometry and streaming apoptosis assay were used to detect the proliferation, cloning formation, cell cycle distribution and the rate of apoptosis in each group, respectively. Results: IHC showed the positive expression rate of Bcl-3 protein in cancer tissues and adjacent normal tissues was 72.09% and 48.84% (P<0.05), respectively. The Bcl-3 protein expression in colorectal cancer tissues was positively correlated with histological grade, lymph node metastasis and Dukes staging (P<0.05). After being transfected for 48 h, the relative Bcl-3 expression level of blank control group and control siRNA group was significantly higher than that of Bcl-3 siRNA group (P<0.05), indicating successful transfection. MTT assay showed the absorbance value in all groups increased in a time-dependent manner. The optical density of Bcl-3 siRNA group was significantly lower than that of blank control group and control siRNA group at 24, 48, 72 and 96 h after transfection (P<0.05), and colony formation rate of Bcl-3 siRNA group was obviously lower than the other two groups (P<0.05). Flow cytomety showed that the proportion of G2 phase cells and cell apoptosis rate of Bcl-3 siRNA group was higher than those of blank control group and the control siRNA group (P<0.05). Conclusions: Bcl-3 protein expression in colorectal cancer tissues was positively correlated with histological grade, lymph node metastasis and Dukes staging; Bcl-3silencing might induce HCT 116 cell cycle arrest at G phase, suppress cell proliferation and colony formation, and promote cell apoptosis.
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Project Supported:
Project supported by the Suzhou Youth Program of Science and Education （No.KJXW2015053），and the Kunshan Science and Technology Program of Social Development（No.KS1654）
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