Expression of HOXA13 in non-small cell lung cancer tissues and its effect on growth of xenograft in nude mice
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Abstract:
[Abstract] Objective: To investigate the expression of HOXA13 in non-small cell lung cancer (NSCLC) tissues, and to explore the effect of silencing HOXA13 gene on the growth of A549 cells in vitro and xenograft in nude mice. Methods: A total of 112 pairs of NSCLC tissues and corresponding adjacent normal tissues from patients, who underwent surgical resection at the Department of Thoracic surgery, Central Hospital of Jiaozuo Coal Industry Group from March 2014 to April 2016, were selected for this study. Real-time fluorescence quantitative PCR was used to detect the expressions of HOXA13 in NSCLC tissues and adjacent tissues. A549 cells were cultured and divided into siRNA-HOXA13 group, negative control group and control group. Real-time quantitative PCR was used to detect the expression of HOXA13 in cells, CCK-8 was used to detect cell proliferation, and Transwell assay was used to detect cell invasion.Xenograft model in nude mice was constructed, and the growth of nude mice was observed. After 5 weeks, the mice was sacrificed and weighed, and the tumor-inhibition rate was calculated. Real-time fluorescence quantitative PCR(qPCR) was used to detect the expression of HOXA13 in xenograft tissues. Results: The relative expression level of HOXA13 mRNA in NSCLC tissues was significantly higher than that in the adjacent tissues (1.83±0.13 vs 1.12±0.10, t=47.008, P=0.000), and its expression was correlated to TNM staging,differentiation and lymph node metastasis (all P<0.05). The relative expression level of HOXA13 mRNA in cells of siRNAHOXA13 group was lower than that in the negative control group and the control group (P<0.05). The cell proliferation level (D values)at 24, 48, 72 and 96 h in the siRNA-HOXA13 group were significantly lower than those in the negative control group and control group (F=30.727, 5.427, 13.816 and 24.454, all P<0.05 or P<0.01); the number of invasive cells in the siRNA-HOXA13 group was lower than that in the negative control group and the control group (all P<0.05). The mass of xenograft in nude mice at week 5 in the siRNA-HOXA13 group was smaller than that in the negative control group and control group, while the tumor-inhibition rate was higher than the negative control group (all P<0.05). The relative mRNA expression level of HOXA13 in xenograft tumor tissue in the siRNA-HOXA13 group was lower than that in the negative control group and the control group (all P<0.01). Conclusion: HOXA13 was highly expressed in the non-small cell lung cancer tissues, and was related to oncogenesis, progression and metastasis of cancer. Specific silencing of HOXA13 gene expression could inhibit the proliferation and invasion of tumor cells and suppress the growth of xenograft in nude mice.
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Project supported by the Science and Technology Development Plan of Henan Province (No.152300410158)