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. 2015 Aug;10(2):990-994.
doi: 10.3892/ol.2015.3303. Epub 2015 Jun 2.

Upregulation of SOX9 promotes cell proliferation, migration and invasion in lung adenocarcinoma

Xiaoying Wang  1 , Ying Ju  2 , M I Zhou  3 , Xiaoli Liu  3 , Chengjun Zhou  1
Affiliations

Upregulation of SOX9 promotes cell proliferation, migration and invasion in lung adenocarcinoma

Xiaoying Wang et al. Oncol Lett. 2015 Aug.

Abstract

Sex determining region Y-box 9 (SOX9) is an important transcription factor in development and has been implicated in several types of cancer. Although the association between upregulation of SOX9 and lung adenocarcinoma (ADC) has been reported previously, the role of SOX9 in the proliferation, migration and invasion of cancer cells remains unclear. Therefore, in the present study, SOX9 expression was detected in 163 human lung adenocarcinoma tissues by immunohistochemistry and western blotting. It was found that the SOX9 protein was over-expressed in the majority of lung adenocarcinoma. The full-length human SOX9 plasmid was then transfected into the lung ADC A549 cell line. An MTT assay was used to investigate the role of SOX9 in cell proliferation. Scratch and extracellular matrix cell invasion assays were performed to investigate whether SOX9 promotes the migration and invasion of lung ADC cells. The results revealed that ectopic overexpression of SOX9 in the lung adenocarcinoma cell line resulted in a marked increase in cell proliferation, migration and invasion. Accordingly, knockdown of SOX9 by RNA interference resulted in the inhibition of cell growth, migration and invasion. The present data indicate that SOX9 may act as a novel marker for lung adenocarcinoma and perform an important role in cell proliferation, migration and invasion.

Keywords: SOX9; invasion; lung adenocarcinoma; migration; proliferation.

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Figures

Figure 1.
Figure 1.
SOX9 protein expression in lung ADC samples and adjacent non-tumorous lung tissues. SOX9 expression was detected by immunohistochemistry. (A) Weak SOX9 expression was observed in all adjacent non-tumorous lung tissues. By contrast, the majority of lung ADC tissues demonstrated (B) moderate or (C) strong SOX9 expression (P<0.01). Magnifiction, x100. (D) The western blot analysis also revealed that the protein level of SOX9 was upregulated in cancerous tissues compared with the corresponding non-cancerous controls (P<0.01). SOX9, sex determining region Y-box 9; ADC, adenocarcinoma.
Figure 2.
Figure 2.
SOX9 is important for the proliferation of lung ADC cells. (A) The A549 cells were transfected with the full-length human SOX9 plasmid and the overexpression of SOX9 was confirmed by western blot analysis. (B) MTT assay revealed that the monolayer cell growth of A549 was significantly increased when SOX9 expression was upregulated. (C) SOX9 expression was also knocked down by RNA interference and the silencing of SOX9 was confirmed by western blotting. (D) An MTT assay revealed that knockdown of SOX9 resulted in a significant decrease of growth capability. ADC, adenocarcinoma; SOX9, sex determining region Y-box 9; siSOX9, small interfering RNA against SOX9; SOX9transfect, cells transfected with the SOX9 plasmid.
Figure 3.
Figure 3.
SOX9 is involved in the regulation of cell migration and invasion in the lung ADC cell line. (A) The A549 cells were transfected with the full-length human SOX9 plasmid or negative control plasmid and incubated for 2 days. The overexpression of SOX9 was confirmed by western blot analysis. (B) The migratory ability of A549 cells was promoted by overexpression of SOX9. The relative migration rates of A549 cells were determined by dividing the migration distance by the time. The migration rate of control cells was defined as 1. Statistical analysis was performed using Student's t-test. P<0.05 was considered to indicate a statistically significant difference. (C) The invasive ability of the A549 cells was promoted by overexpression of SOX9. The results were expressed as relative values, using control cells as 1. (D) The A549 cells were treated with SOX9 siRNA or non-targeting control siRNA and incubated for 2 days. The silencing of SOX9 was confirmed by western blotting. (E) The migratory ability of A549 cells was suppressed as a result of SOX9 knockdown. The results were expressed as the relative values using cells treated with non-targeting control siRNA as 1. (F) The invasive ability of A549 cells was suppressed as a result of SOX9 knockdown. The results were expressed as relative values using cells treated with non-targeting control siRNA as 1. *P<0.05. SOX9, sex determining region Y-box 9; siRNA, small interfering RNA; SOX9transfect, cells transfected with the SOX9 plasmid; siSOX9, siRNA against SOX9.
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