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. 2019 Jan;17(1):113-118.
doi: 10.3892/ol.2018.9613. Epub 2018 Oct 24.

Basic transcription factor 3 expression silencing attenuates colon cancer cell proliferation and migration in vitro

Xu Li  1 , Jinke Sui  1 , Junjie Xing  1 , Fuao Cao  1 , Hao Wang  1 , Chuangang Fu  1 , Hantao Wang  1
Affiliations

Basic transcription factor 3 expression silencing attenuates colon cancer cell proliferation and migration in vitro

Xu Li et al. Oncol Lett. 2019 Jan.

Abstract

Basic transcription factor 3 (BTF3) is an RNA polymerase II transcription factor that also regulates apoptosis. Numerous studies have identified that BTF3 is aberrantly expressed in several types of tumor. However, the function of BTF3 in colorectal cancer remains unknown. The aim of the present study was to assess the function of BTF3 during colon cancer tumorigenesis. Applying a lentivirus-transfected short hairpin RNA approach, expression of BTF3 was dysregulated in the colon cancer HCT116 and HT-29 cell lines; knockdown efficiency was verified using the quantitative polymerase chain reaction and western blotting. To determine the function of BTF3 in colon cancer, cell proliferation was assessed using an MTT assay, cell apoptosis and the cell cycle were assessed using flow cytometry, and cell migration was assessed using a Transwell assay. Knockdown of BTF3 inhibited cell proliferation, possibly because BTF3 knockdown induced cell early apoptosis and arrested cells in G0-G1 phase. BTF3 knockdown also inhibited cell migration. The results of the present study identified that BTF3 expression is associated with colon cancer progress, and BTF3 may therefore be a molecular marker for diagnosis and treatment outcomes of human colon cancer.

Keywords: apoptosis; basic transcription factor 3; cell cycle; colon cancer; migration; proliferation.

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Figures

Figure 1.
Figure 1.
shRNA BTF3 knockdown in colon cancer HCT116 and HT-29 cells. (A) Quantitative polymerase chain reaction analysis detected BTF3 mRNA levels in empty vector control, shRNA-BTF3 lentivirus-transfected, and non-transfected HCT116 (top) and HT-29 (bottom) cells. (B) Western blot analysis of BTF3 protein expression in empty vector control and shRNA-BTF3 lentivirus-transfected, and in non-transfected HCT116 (top) and HT-29 (bottom) cells. β-actin was used as loading control. (C) Semi-quantitative densitometry analysis of BTF3 protein levels in HCT116 (top) and HT-29 (bottom) cells. **P<0.01. NT, non-transfected; shRNA, short hairpin RNA; BTF3, basic transcription factor 3; shBTF3, short hairpin BTF3.
Figure 2.
Figure 2.
Knockdown of BTF3 inhibits cell proliferation in colon cancer cells. The proliferation of (A) HCT116 and (B) HT-29 cells following lentiviral transfection with empty vector control or shBTF3 was assessed using an MTT assay. **P<0.01. shRNA, short hairpin RNA; BTF3, basic transcription factor 3; OD, optical density.
Figure 3.
Figure 3.
BTF3 knockdown induces HCT116 cells early apoptosis and arrested cells in G0-G1 phase of the cell cycle. (A) Flow cytometric analysis of empty vector control, shBTF3-transfected cells and non-transfected HCT116 cells apoptosis using annexin V and PI double staining. The proportion of early apoptotic cells (annexin-V-positive and PI-negative) was quantified. (B) Cell cycle distribution of empty vector control, shBTF3-transfected or non-transfected HCT116 cells determined by flow cytometry. **P<0.01. BTF3, basic transcription factor 3; shRNA, short hairpin RNA; NT, non-transfected; PI, propidium iodide; shBTF3, short hairpin BTF3; Dip, diploid peak.
Figure 4.
Figure 4.
BTF3 knockdown inhibits human colon cancer cell motility. (A) The migratory ability of human colon cancer HCT116 (top) and HT-29 (bottom) cell lines transfected with an empty vector control or with shBTF3 was assessed using a Transwell assay, as was that of non-transfected cells. (B) Quantification of migration indices. Each experiment was performed in triplicate. **P<0.01. BTF3, basic transcription factor 3; shRNA, short hairpin RNA; NT, non-transfected; shBTF3, short hairpin BTF3.
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